A Phase I Study of Immunotherapy With E6 T Cell Receptor T Cells for Vulvar High-Grade Squamous Intraepithelial Lesions
Phase: PHASE1
Conditions: Human Papillomavirus, HPV-16, High Grade Squamous Intraepithelial Lesion
Sponsor: National Cancer Institute (NCI)

Abbreviated Title: Immunotherapy for vulvar HSIL
Version Date: June 13, 2018
1
Abbreviated Title: Immunotherapy for vulvar HSIL
CC Protocol #: 17-C-0116 B
OSP #: 1704-1606
IBC #: RD-17-III-09
NCT #: NCT03197025
Version Date: June 13, 2018
Title: A Phase I Study of Immunotherapy with E6 T Cell Receptor T Cells for Vulvar High-
Grade Squamous Intraepithelial Lesions
Principal Investigator: Christian S. Hinrichs, MD
Experimental Transplantation and Immunology Branch
National Cancer Institute (NCI)
10 Center Drive, Room 4B04
Bethesda, MD 20892
Phone: 240-760-6059
Email: Hinrichs@nih.gov
Investigational Agents: 
Drug Name:
E6 TCR
IND Number:
16078
Sponsor:
Center for Cancer Research 
Manufacturer:
CC DTM
Commercial Agents:
Interleukin-2

Abbreviated Title: Immunotherapy for vulvar HSIL
Version Date: June 13, 2018
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PRÉCIS 
Background:
x
Vulvar high-grade squamous intraepithelial lesion (HSIL) is a premalignant epithelial
lesion that is frequently multifocal and/or recurrent.
x
The primary treatment is surgery, which may result in disfigurement and compromise of
the urethra, anus, or clitoris. Recurrence after surgery is common and primarily treated with
additional surgery.
x
Vulvar HSIL is caused by chronic infection with the human papillomavirus (HPV) type 16
infection. In this clinical trial the HPV-16 infection is targeted with a single infusion of
autologous T cells that have been genetically engineered to express an HPV-16 E6-specific
T cell receptor (E6 TCR T cells).
Objective:
x
Determine the safety of E6 TCR T cells for the treatment of vulvar HSIL.
Eligibility:
x
Histologically confirmed diagnosis of HPV-16+ vulvar HSIL.
x
Expression of the HLA-A2*02:01 allele.
x
Measurable lesion(s) that are recurrent or cannot be resected with acceptable cosmetic or
functional results.
x
Age greater than or equal to 18 years old and less than or equal to 65 years old.
x
Eastern Oncology Cooperative Group Performance Score of 0 or 1.
Design:
x
This is a phase I clinical trial with a 3+3 dose escalation design.
x
Subjects will receive E6 TCR T cells followed by up to two doses of aldesleukin 720,000
IU/kg IV.
x
No conditioning regimen will be given, aldesleukin will be capped at a maximum of two
doses, and E6 TCR T cell dosing will begin at dose level -1 from the previously determined
safe dose.

Abbreviated Title: Immunotherapy for vulvar HSIL
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TABLE OF CONTENTS 
PRÉCIS........................................................................................................................................... 2 
TABLE OF CONTENTS................................................................................................................ 3 
1. 
INTRODUCTION .................................................................................................................. 8 
1.1 
Study Objectives .............................................................................................................. 8 
1.1.1 
Primary Objective..................................................................................................... 8 
1.1.2 
Secondary Objectives................................................................................................ 8 
1.1.3 
Exploratory Objectives ............................................................................................. 8 
1.2 
Background and Rationale ............................................................................................... 8 
1.2.1 
Vulvar high-grade squamous intraepithelial lesion (HSIL)...................................... 8 
1.2.2 
Studies of the natural history and treatment history of vulvar HSIL........................ 9 
1.2.3 
Targeting of HPV-associated cancers with adoptive T cell therapy......................... 9 
1.2.4 
E6 TCR discovery and characterization ................................................................. 10 
1.2.5 
High-dose Interleukin-2 (Aldesleukin)................................................................... 13 
1.2.6 
Safety Considerations ............................................................................................. 13 
2 
ELIGIBILITY ASSESSMENT AND ENROLLMENT....................................................... 14 
2.1 
Eligibility Criteria .......................................................................................................... 14 
2.1.1 
Inclusion Criteria .................................................................................................... 14 
2.1.2 
Exclusion Criteria ................................................................................................... 16 
2.2 
Screening Evaluation...................................................................................................... 16 
2.2.1 
Any time prior to starting the therapeutic regimen:................................................ 16 
2.2.2 
Within 4 weeks prior to starting the therapy unless otherwise indicated: .............. 17 
2.2.3 
Within 14 days prior to starting the therapy: .......................................................... 17 
2.2.4 
Within 7 days prior to starting the therapy regimen:.............................................. 17 
2.3 
Registration Procedures.................................................................................................. 17 
2.3.1 
Treatment Assignment and Randomization/Stratification Procedures for 
registration purposes only..................................................................................................... 18 
2.4 
Baseline Procedures ....................................................................................................... 18 
3 
STUDY IMPLEMENTATION ............................................................................................ 19 
3.1 
Study Design .................................................................................................................. 19 
3.1.1 
Protocol schema...................................................................................................... 20 
3.1.2 
Leukapheresis ......................................................................................................... 21 
3.1.3 
E6 TCR T cell preparation...................................................................................... 21 

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3.1.4 
Treatment Phase...................................................................................................... 21 
3.1.5 
Dose Limiting Toxicities (DLTs) ........................................................................... 22 
3.1.6 
Dose Escalation....................................................................................................... 22 
3.1.7 
Safety Protocol Stopping Rules .............................................................................. 22 
3.2 
Drug Administration ...................................................................................................... 23 
3.3 
Study Calendar............................................................................................................... 24 
3.4 
On-Study Evaluations .................................................................................................... 26 
3.4.1 
Prior to starting cell infusion: ................................................................................. 26 
3.4.2 
After Cell Infusion:................................................................................................. 27 
3.4.3 
During Hospitalization:........................................................................................... 27 
3.5 
Post Treatment Evaluation (Follow-Up)........................................................................ 27 
3.5.1 
Evaluation:.............................................................................................................. 27 
3.5.2 
At each Response Evaluation, patients will undergo the following: ...................... 28 
3.5.3 
Patients who are unable or unwilling to return for follow up evaluations.............. 28 
3.5.4 
Long-term Follow-Up:............................................................................................ 28 
3.6 
Criteria for Removal from Protocol Therapy and Off Study Criteria............................ 29 
3.6.1 
Criteria for removal from protocol therapy ............................................................ 29 
3.6.2 
Off-Study Criteria................................................................................................... 29 
3.6.3 
Off Protocol Therapy and Off-Study Procedure..................................................... 29 
4 
CONCOMITANT MEDICATIONS/MEASURES.............................................................. 30 
4.1 
Steroids........................................................................................................................... 30 
4.1.1 
Blood Product Support............................................................................................ 30 
4.2 
Other concomitant medications/measures...................................................................... 30 
5 
BIOSPECIMEN COLLECTION.......................................................................................... 30 
5.1 
Correlative Studies for Research.................................................................................... 30 
5.1.1 
Pre cell infusion evaluations ................................................................................... 30 
5.1.2 
Post cell infusion evaluations.................................................................................. 31 
5.1.3 
Optional vulvar Biopsies......................................................................................... 31 
5.1.4 
Immunological Testing........................................................................................... 32 
5.2 
Summary of Sample Collection ..................................................................................... 32 
5.3 
Gene-therapy-specific follow-up.................................................................................... 33 
5.4 
Sample Storage, Tracking and Disposition.................................................................... 33 

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5.4.1 
Storage/Tracking in the Preclinical Development and Clinical Monitoring Facility 
(PDCMF) .............................................................................................................................. 33 
5.4.3 
Storage/Track in the Christian Hinrichs lab............................................................ 34 
5.4.4 
Storage/Tracking in the Liang Cao lab................................................................... 35 
5.4.4 
Protocol Completion/Sample Destruction .............................................................. 35 
6 
DATA COLLECTION AND EVALUATION..................................................................... 35 
6.1 
Data Collection............................................................................................................... 35 
6.1.1 
Exclusions to Routine Adverse Event Reporting: .................................................. 36 
6.2 
Data Sharing Plans ......................................................................................................... 36 
6.2.1 
Human Data Sharing Plan....................................................................................... 36 
6.3 
Response Criteria ........................................................................................................... 37 
6.3.1 
Definitions............................................................................................................... 37 
6.3.2 
Disease Parameters ................................................................................................. 37 
6.3.3 
Methods for Evaluation of Measurable Disease..................................................... 37 
6.3.4 
Response Criteria.................................................................................................... 37 
6.3.5 
Duration of Response.............................................................................................. 39 
6.4 
Toxicity Criteria............................................................................................................. 39 
7 
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING PLAN
39 
7.1.1 
Adverse Event......................................................................................................... 39 
7.1.2 
Suspected adverse reaction ..................................................................................... 40 
7.1.3 
Unexpected adverse reaction .................................................................................. 40 
7.1.4 
Serious..................................................................................................................... 40 
7.1.5 
Serious Adverse Event............................................................................................ 40 
7.1.6 
Disability................................................................................................................. 40 
7.1.7 
Life-threatening adverse drug experience............................................................... 40 
7.1.8 
Protocol Deviation (NIH Definition)...................................................................... 40 
7.1.9 
Non-compliance (NIH Definition).......................................................................... 41 
7.1.10 Unanticipated Problem............................................................................................ 41 
7.2 
NCI-IRB and Clinical Director Reporting ..................................................................... 41 
7.2.1 
NCI-IRB and NCI Clinical Director Expedited Reporting of Unanticipated 
Problems and Deaths............................................................................................................. 41 
7.2.2 
NCI-IRB Requirements for PI Reporting at Continuing Review........................... 41 

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7.2.3 
NCI-IRB Reporting of IND Safety Reports............................................................ 42 
7.3 
IND Sponsor Reporting Criteria .................................................................................... 42 
7.3.1 
Reporting Pregnancy/Maternal exposure .............................................................. 42 
7.4 
Institutional Biosafety Committee (IBC) Reporting Criteria......................................... 43 
7.4.1 
Serious Adverse Event Reports to IBC................................................................... 43 
7.4.2 
Annual Reports to IBC............................................................................................ 43 
7.5 
Data and Safety Monitoring Plan................................................................................... 44 
7.5.1 
Principal Investigator/Research Team.................................................................... 44 
7.5.2 
Sponsor Monitoring Plan........................................................................................ 44 
7.5.3 
Safety Monitoring Committee (SMC) .................................................................... 45 
8 
STATISTICAL CONSIDERATIONS.................................................................................. 45 
9 
COLLABORATIVE AGREEMENTS................................................................................. 45 
9.1 
Cooperative Research and Development Agreement (CRADA)................................... 45 
10 HUMAN SUBJECTS PROTECTIONS ............................................................................... 45 
10.1 Rationale For Subject Selection..................................................................................... 45 
10.2 Strategies/Procedures for Recruitment........................................................................... 46 
10.3 Participation of Children................................................................................................ 46 
10.4 Participation of Subjects Unable to Give Consent......................................................... 46 
10.5 Evaluation of Benefits and Risks/Discomforts .............................................................. 46 
10.6 Risks/Benefits Analysis (including who are or may become unable to consent) .......... 46 
10.7 Consent Process and Documentation............................................................................. 47 
10.7.1 Telephone consent and re-consent.......................................................................... 47 
10.7.2 Enrollment of non-English Speaking Subjects ....................................................... 47 
11 PHARMACEUTICAL INFORMATION............................................................................. 48 
11.1 Interleukin-2 (Aldesleukin, Proleukin, Recombinant Human Interleukin 2)................. 48 
11.1.1 How Supplied: ........................................................................................................ 48 
11.1.2 Formulation/Reconstitution: ................................................................................... 48 
11.1.3 Storage: ................................................................................................................... 48 
11.1.4 Dilution/Stability: ................................................................................................... 48 
11.1.5 Administration: ....................................................................................................... 48 
11.1.6 Toxicities: ............................................................................................................... 48 
11.2 Cell Preparation (E6 TCR Transduced PBL) (IND # 16078)........................................ 49 
11.2.1 Retroviral Vector Containing the E6 TCR gene..................................................... 49 

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12 REFERENCES ..................................................................................................................... 50 
13 APPENDICES ...................................................................................................................... 52 
13.1 Appendix A -Performance Status Criteria ..................................................................... 52 
13.2 $SSHQGL[%$GYHUVH(YHQWV2FFXUULQJ,Q2I3DWLHQWV7UHDWHG:LWK$OGHVOHXNLQ
(N=525)1 ................................................................................................................................... 53 
13.3 Appendix C: Expected Aldesleukin Toxicities and Their Management........................ 54 
13.4 Appendix D: Certificate of Analysis HPV-16 E6 TCR ................................................. 56 
13.5 Appendix E:  HPV-16 DDPCR Peripheral Blood Processing Instructions to Patient and 
Physician................................................................................................................................... 58 

Abbreviated Title: Immunotherapy for vulvar HSIL
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1. INTRODUCTION
1.1
STUDY OBJECTIVES
Objectives:
1.1.1
Primary Objective
x
Determine the safety of E6 TCR T cells for the treatment of vulvar HSIL.
1.1.2
Secondary Objectives
x
Assess clinical responses in subjects treated with E6 TCR T cells for vulvar HSIL.
x
To evaluate a novel assay to genotype HPV from metastatic HPV+ cancers using a
ddPCR-based blood test.
1.1.3
Exploratory Objectives
ƒ
Test for clearance of HPV infection from vulvar tissues.
ƒ
Evaluate the expansion, survival, trafficking, phenotype, and function of E6 
TCR cells following infusion.
ƒ
Investigate the vulvar HSIL microenvironment including immune cell subsets 
and costimulatory/inhibitory molecules.
ƒ
Assess circulating HPV DNA levels associated with treatment. 
1.2
BACKGROUND AND RATIONALE
Vulvar high-grade squamous intraepithelial lesion (HSIL) is a premalignant condition that is
caused by high-risk human papillomavirus (HPV) infection. The primary treatment for vulvar
HSIL is surgical. Surgery can result in substantial morbidity, particularly for lesions that are
multifocal, extensive, or adjacent to important anatomic structures (e.g. the anus, clitoris, or
urethra). Following surgery, disease recurrence is common due to persistence of the underlying
HPV infection. Therefore, a systemic treatment directed against the viral infection at the
foundation of the disease may be more effective. This research protocol seeks to target the HPV
infection that is causing vulvar HSIL with immunotherapy. The immunotherapy consists of
autologous T cells that are genetically engineered to recognize the HPV-16 E6 oncoprotein,
which is expressed by HPV-16 infected cells. The protocol is designed as a one-time treatment to
clear the HPV infection and thereby cause regression of the HSIL lesions.
1.2.1
Vulvar high-grade squamous intraepithelial lesion (HSIL)
Vulvar HSIL is defined by the 2015 International Society for the Study of Vulvovaginal Disease
(ISSVD) as an HPV-associated lesion of the vulva that has high malignancy potential. The term,
vulvar HSIL, is equivalent to the previously used nomenclature of vulvar intraepithelial
neoplasia, usual type [1]. It does not include low-grade squamous intraepithelial lesions without
malignant potential such as condyloma acuminatum. It also does not include premalignant
lesions that are not associated with HPV, which are classified as differentiated-type vulvar
intraepithelial neoplasia. Multiple HPV types can cause vulvar HSIL but HPV-16 is the most
prevalent [2]. Spontaneous regression of vulvar HSIL is rare, reportedly occurring in fewer than
1.5% of patients per year [3].

Abbreviated Title: Immunotherapy for vulvar HSIL
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Patients with vulvar HSIL may present with symptoms of local itching, burning, or pain.
Diagnosis is suggested by careful inspection of the lesion(s) and is confirmed by biopsy.
Multicentric disease is common due to diffuse spread of HPV infection throughout vulvar
epithelium. Vulvar HSIL transforms into vulvar squamous cell carcinoma (SCC) in 3 to 9% of
patients. Therefore, all lesions require surveillance with physical exams and biopsies to rule out
invasive cancer [4]. For lesions that do not regress, excision is required. The type of surgery
depends on the extent and anatomic location of disease. Treatment of multifocal lesions,
extensive disease, or lesions in proximity to the urethra, clitoris, or anus can result in deformity
and functional impairment. Even with radical excision to negative margins, recurrence occurs in
approximately 30% of patients [5]. Thus, surgery may be an unsatisfactory treatment option as it
often fails to prevent disease recurrence, results in morbidity, and negatively impact
psychological and sexual health [6].
1.2.2
Studies of the natural history and treatment history of vulvar HSIL
Studies of the natural history and treatment of vulvar HSIL are scientifically limited as they are 
mostly retrospective and have small sample sizes. One systematic review reported that 6.5% of 
3322 patients with vulvar HSIL progressed to develop an invasive vulvar carcinoma [3] . In 
untreated patients, invasive vulvar carcinoma developed in nine percent of subjects within one to 
eight years. Invasive cancers are treated with radical local excision or modified radical 
vulvectomy, depending on the tumor stage. Locally advanced cancers are treated with 
chemoradiation. Salvage surgery with pelvic exenteration and colostomy or ureteral diversion 
may be employed. Five-year survival for local, regional, and distant vulvar cancers are 86%, 
54%, and 16% respectively. A study based on data from the United States reported the incidence 
of vulvar HSIL to be 2.86. per 100,000 women [7]. Immunotherapy for vulvar HSIL
Vulvar HSIL is caused by HPV infection, and HPV infections can be recognized and attacked by
the immune system. Hence, immunotherapy is an attractive treatment strategy for vulvar HSIL.
Topical immunotherapy with imiquimod, an immune response modifier that activates dermal
dendritic cells through Toll-like receptor 7, has been reported to have response rates of 13-81%,
but vulvar inflammation and pain limit its use and it is not currently FDA-approved [8].
Nonetheless, it is a treatment option that some patients should consider as an alternative to this
protocol. Experimental vaccines have been tested for the treatment of vulvar HSIL. A clinical
trial of a vaccine targeting HPV-16 E6 and E7 showed promising results in a single-arm phase II
study [9]. This vaccine is experimental and not available outside of a clinical trial. The
preventative vaccines for HPV (e.g. Gardasil and Cervarix) are highly effective at preventing
HPV infections but not considered effective for the treatment of VIN [10].
1.2.3
Targeting of HPV-associated cancers with adoptive T cell therapy
In a clinical trial of HPV-16 E6 TCR T cells for patients with metastatic cancers associated with
HPV-16 (e.g. cervical cancer, anal cancer, head and neck cancer, etc.), subjects received a
chemotherapy conditioning regimen followed by E6 TCR T cells and aldesleukin (protocol 15-
C-0005). Two of 12 patients experienced objective tumor responses. No E6 TCR T cell dose-
limiting toxicities, off-target toxicity, healthy tissue targeting, or cytokine storm were observed.
The maximum administered dose was deemed the maximum tolerated dose. Separately, in a
clinical trial of autologous tumor-infiltrating T cells generated from cultures with HPV reactivity
for metastatic HPV+ cancers, objective tumor responses occurred in 3/9 patients with cervical

Abbreviated Title: Immunotherapy for vulvar HSIL
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10
cancer [11]. Adoptive T cell therapy has not been reported for the treatment of premalignant
HPV-related diseases.
1.2.4
E6 TCR discovery and characterization
HPV-16 E6 is a viral oncoprotein that is constitutively expressed by and important to the 
survival of HPV-16+ infected cells, and it is absent from healthy human tissues. We isolated a 
TCR from the tumor-infiltrating T cells of a patient with metastatic anal cancer that recognized
the E629-38 epitope (described below).
Figure 1. PG13-oDCA2-E6-C12 TCR vector map. An MSGV1 retroviral vector encoding the
TCR was constructed. This retroviral vector consists of 7,325 base pairs and includes a 5’LTR 
from the murine stem cell virus (promoter), packaging signal including the splicing donor (SD) 
and splicing acceptor sites (SA). Alpha and beta chains of the E6 TCR are linked by a P2A 
peptide.
Peripheral blood T cells transduced to express the E6 TCR displayed high avidity for the E629-38
peptide (Figure 2) and CD8-independent HLA-A*02:01/E629-38 tetramer binding (Figure 3).
They specifically recognized a panel of HPV-16+ HLA-A*02:01+ cervical and oropharyngeal 
cancer cell lines. Thus, TCR gene engineered T cells expressing the E6 receptor can be used to 
target HPV-16+ HLA-A*02:01+ cells.

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Figure 2. T cells transduced to express the E6 TCR demonstrated high avidity for the E629- 38 
peptide. CD4 and CD8 T cells from PBMC were transduced to express the E6 TCR. Functional 
avidity was tested by coculture with T2 cells pulsed with titrated concentrations of E629-38 
peptide.

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HLA-A2/E629-38 
mTRBC
Figure 3. Peripheral blood T cells transduced to express the E6 TCR display CD8-
independent HLA-A*02:01/E629-38 tetramer binding. T cells from PBMC were transduced to 
express the E6 TCR. Dot plots shown are gated on PI- lymphocytes.
In contrast to TCRs that have had unexpected cross-reactivity against normal human proteins, 
this TCR was isolated directly from a human T cell. Hence, it was subjected to thymic selection 
and is unlikely to possess avid reactivity against self-antigens. The complementarity determining 
regions of the TCR have not been modified, so there is no chance that cross-reactivity has been
artificially introduced. The target epitope is derived from a viral protein, and no more than 6 of 
its 10 amino acids are shared with any human protein. There is no cross reactivity of this TCR 
against epitopes of these human proteins (Figure 4).
HPV-16E629-38
ITGA8
DMBX1
ACO2
PTPRG
HPTPG
PTPRU
no peptide
T2 alone
Tcell alone
OKT3
0
10000
20000
30000
IFN-J (pg/mL)

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Gene Abbreviation 
Sequence 
Full Name of Protein 
HPV16 E629-38 
TIHDIILECV
HPV-16 E629-38 
ITGA8 
TISDTILEVG
Integrin alpha-8 
DMBX1 
TFQDIILEAR
Diencephalon/mesencephalon homeobox 1 
ACO2 
IIHQIILENY
Aconitate hydratase, mitochondrial 
PTPRG 
FIHDFILEAI
Protein tyrosine phosphatase 
HPTPG 
IIHDFILEAT
Receptor-type tyrosine-protein phosphatase gamma 
PTPRU 
FIHDAILEAC
Receptor-type tyrosine-protein phosphatase kappa or mu or U 
Figure 4(7&5WUDQVGXFHG7FHOOVGLGQRWUHFRJQL]H7FHOOVSXOVHGZLWKSHSWLGHVVKDULQJ
6 amino acids with peptides from human proteins identified by BLAST search.  T2 cells pulsed 
with the peptides indicated in the table were cocultured with T cells expressing the E6 TCR.  
1.2.5
High-dose Interleukin-2 (Aldesleukin)
Interleukin-2 (IL-2) is a T cell growth factor that has been administered as a single agent and in
combination with adoptive T cell therapy for the treatment of cancer [12]. As a single agent it
can induce regression of metastatic melanoma and renal cell carcinoma with overall response
rates around 20% [13]. Although high quality published data comparing adoptive T cell therapy
with and without IL-2 are lacking, data from animal models indicate that IL-2 enhances the anti-
tumor function of adoptively transferred T cells [14, 15].
Single-agent high-dose aldesleukin for invasive cancers is administered at a fixed dose (600,000
or 720,000 IU/Kg) but a variable number of doses are given depending on individual tolerance of
the drug. Aldesleukin infusions are continued until adverse events prohibit further drug
administration. To increase that safely of the present protocol, aldesleukin dose number will be
capped at two doses, a number of doses that has been well tolerated in prior studies [16, 17]. In
prior studies, the toxicity of aldesleukin was not increased by coadministration of autologous T
cells [18, 19].
1.2.6
Safety Considerations
HPV-16 E6 TCR T cells target a foreign viral protein that is not present in human tissue. The
TCR was discovered from the study of tumor infiltrating T cells isolated from a human and
therefore has undergone thymic selection to not have human autoreactivity. The nucleotide
sequence of the E6 TCR was codon optimized for expression in human tissues and the TCR
constant regions replaced with their murine counterparts to improve TCR alpha/beta chain
pairing and TCR expression. The alpha/beta TCR sequence was cloned into the MSGV1
retroviral vector, which has been tested extensively in clinical trials. TCR avidity enhancement
was not performed and wild-type CDR regions are present, therefore there is no chance that off-
target reactivity has been artificially introduced. The clinical safety of E6 TCR T cells has been
studied in a protocol for the treatment of metastatic HPV-16 cancers (15-C-0005). In that study,

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a conditioning chemotherapy regimen that enhances T cell survival and function, and increases
the toxicity of the treatment was employed, and high-dose aldesleukin was administered to
individual patient tolerance. Twelve patients were treated and no dose-limiting toxicity for E6
TCR T cell infusion was encountered. The E6 TCR T cells did not demonstrate off-target
toxicity, targeting of healthy tissues, or cytokine storm. The maximum administered dose was
thus declared to be the maximum tolerated dose (1.7 x 1011 cells).
In the present protocol, the safety margin of the treatment has been increased by the following
measures: 1) No conditioning chemotherapy will be given. 2) Aldesleukin dose number will be
capped at two. 3) E6 TCR T cell dose will begin at dose level -1.
Experience at the NIH Clinical Center treating more than 200 patients with advanced cancers
with genetically engineered T cells indicates that these cells do not have a significant risk of
malignant transformation in this setting. While insertional mutagenesis is a known hypothetical
risk, there has never been a reported vector-mediated gene transfer into mature T cells since the
first NCI sponsored gene transfer study in 1989 [20]. This proposed protocol follows all current
FDA guidelines regarding testing and follow up of patients receiving gene transduced cells.
2
ELIGIBILITY ASSESSMENT AND ENROLLMENT
2.1
ELIGIBILITY CRITERIA
2.1.1
Inclusion Criteria
2.1.1.1 Patients must have vulvar HSIL as confirmed by pathology report from a CLIA-certified
laboratory.
2.1.1.2
Vulvar HSIL must be HPV-16+ by a PCR, RNA, or in situ hybridization test from a 
CLIA certified laboratory.
2.1.1.3
Patients must have measurable lesion(s) as defined in section 6.3.2 and one or more of 
the following criteria:
a.
Failure of surgery to control disease (i.e. positive margins or recurrence of 
HSIL after surgery).
b. Multifocal or extensive disease for which surgery would result in major 
deformity that is not be acceptable to the patient.
c.
Disease for which surgery would have a risk of functional impairment that 
is not be acceptable to the patient (i.e. involve partial or complete excision 
of the clitoris, anus, vagina, or urethra).
2.1.1.4
Patients may have received any previous therapy, including surgical excision, but must
have histologically documented recurrence on new biopsy and a measurable lesion that
meets the above criteria.
2.1.1.5
The presence of disease that can be biopsied for research purposes is not an inclusion
criterion.
2.1.1.6
Patients must have the HLA-A*02:01 allele

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15
2.1.1.7
Age >18 years and  65 years. As age increases, the ability to tolerate the toxicities of
aldesleukin decreases, so the patient population for this study will include up to and
including 65 years of age to increase safety.
2.1.1.8
ECOG performance status of 0-1 as defined by Appendix A. Able to understand and
sign the Informed Consent Document.
2.1.1.9
Women of child-bearing potential must have a negative pregnancy test. Women of
child-bearing potential are defined as all women who are not post-menopausal or who
have not had a hysterectomy. Postmenopausal will be defined as women over the age
of 55 who have not had a menstrual period in at least 1 year.
2.1.1.10 The effects of E6TCR T Cells on the developing human fetus are unknown.  For this 
reason, women of child-bearing potential must agree to use adequate contraception
(hormonal or barrier method of birth control; abstinence) prior to study entry and for
the duration of study participation. Should a woman become pregnant or suspect she is
pregnant while she is participating in this study, she should inform her treating
physician immediately.
2.1.1.11 Seronegative for HIV antibody. The experimental treatment being evaluated in this
protocol depends on an intact immune system. Patients who are HIV seropositive can
have decreased immune-competence and thus be less responsive to the experimental
treatment.
2.1.1.12 Seronegative for hepatitis B antigen and hepatitis C antibody. If hepatitis C antibody
test is positive, then the patient must be tested for the presence of antigen by RT-PCR
and be HCV RNA negative.
2.1.1.13 Must be willing to participate in Gene Therapy Long Term Followup Protocol (15-C-
0141), which will follow patients for up to 15 years per Food and Drug Administration 
(FDA) requirements.
2.1.1.14 Patients must have normal organ and marrow function as defined below:
x
leukocytes
>3,000/mcL
x
absolute neutrophil count
>1,000/mcL
x
platelets
>150,000/mcL
x
hemoglobin
>10.0 g/dL
x
total bilirubin
within normal institutional limits except
in patients with Gilbert’s Syndrome who
must have a total bilirubin < 3.0 mg/dL
x
AST(SGOT)/ALT(SGPT)
Serum ALT/AST < 3X ULN
x
creatinine
< 1.5X baseline, < 1.5X ULN
OR
x
creatinine clearance
>60 mL/min/1.73 m2 for patients with
creatinine levels above institutional normal
(by the Chronic Kidney Disease Epidemiology
Collaboration (CKD-EPI) equation)

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16
2.1.2
Exclusion Criteria
2.1.2.1
Patients who are receiving any other investigational agents
2.1.2.2
Because there is an unknown but potential risk for adverse events in nursing 
infants secondary to treatment of the mother with E6 TCR, breastfeeding should 
be discontinued if the mother is treated with E6 TCR.  These potential risks may 
also apply to other agents used in this study.
2.1.2.3
Uncontrolled intercurrent illness including, but not limited to, any ongoing or 
active infection (e.g. requiring anti-infective therapy), coagulation disorders, 
cardiovascular disorders, respiratory disorders, cancer, or psychiatric illness/social 
situations (within the last six months) that would limit compliance with study 
requirements. 
2.1.2.4
Any form of systemic immunodeficiency, including acquired deficiency such as
HIV or primary immunodeficiency such as Severe Combined Immunodeficiency
Disease. The experimental treatment being evaluated in this protocol depends on
an intact immune system. Patients who have decreased immune competence may
be less responsive to the treatment.
2.1.2.5
Concurrent systemic steroid therapy if greater than the equivalent of 5 mg
prednisone PO daily. Patients previously on steroids must be off steroids for four
weeks prior to treatment.
2.1.2.6
Any history of clinically significant cardiac arrhythmia, coronary
revascularization, ischemic symptoms, or previously documented LVEF of less
than or equal to 45%. A cardiac stress test is required for all patients greater than
50 years old. A cardiac stress test may also be performed for any clinical concern.
Patients with cardiac ischemia are not eligible.
2.1.2.7
Patients with any active invasive cancer are not eligible.
2.1.2.8
Patients vulvar HSIL that is not HPV-16+ or is associated with multiple types of
high-risk HPV are not eligible.
2.2
SCREENING EVALUATION
2.2.1
Any time prior to starting the therapeutic regimen:
a. HLA-Typing
b. Review of most recent cervical cytology and HPV testing. Cervical cytology testing
will be performed if clinically indicated.
c.
If a pathology report confirming vulvar HSIL is not available, if available results are
unclear, or there is a lack of necessary information in the available results, a vulvar
biopsy will be performed to confirm diagnosis.
d. Cervical high-risk HPV testing
e. Venous assessment (as per apheresis clinic policy)
f.
One 10 mL tube of blood will be collected for the optional ddPCR-based research 
blood test.

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17
2.2.2
Within 4 weeks prior to starting the therapy unless otherwise indicated:
a. Complete history and physical examination, noting in detail the exact size and
location of all lesions. (Note: Patient history may be obtained within 8 weeks prior to
starting therapy.)
b. Chest x-ray
c. EKG
d. Cardiac stress test for patients over the age of 50.
e. HIV antibody titer, HbsAG determination, and anti-HCV antibody. (Note: may be
performed within 3 months of therapy start date).
f.
Past medical history including family history, social history, allergies, and current
medications.
2.2.3
Within 14 days prior to starting the therapy:
a.
Chem 20 equivalent: (Sodium (Na), Potassium (K), Chloride (Cl), Total CO2
(Bicarbonate), Creatinine, Glucose, Urea nitrogen (BUN), Albumin, Calcium total,
Magnesium total (Mg), Inorganic Phosphorus, Alkaline Phosphatase, ALT/GPT,
AST/GOT, Total Bilirubin, Direct Bilirubin, LD, Total Protein, Total CK, Uric Acid).
b. Thyroid Panel
c. CBC with differential and TBNK
d. PT/PTT
2.2.4
Within 7 days prior to starting the therapy regimen:
a. Beta-HCG pregnancy test (serum or urine) on all women of child-bearing potential
b. ECOG performance status of 0 or 1
2.3
REGISTRATION PROCEDURES
Registration will be a two-part process as patients are screened on this protocol. Authorized staff 
must register an eligible candidate with NCI Central Registration Office (CRO) within 24 hours 
of signing consent. To initially register a subject after the participant has signed the consent, 
complete the top portion of the registration Eligibility Checklist from the website 
(http://home.ccr.cancer.gov/intra/eligibility/welcome.htm) indicating that the patient is being 
registered for screening and send via encrypted email to: NCI Central Registration Office  
ncicentralregistration-l@mail.nih.gov. Once eligibility is confirmed after completion of 
screening studies, complete the remainder of the form which is the eligibility checklist, 
indicating that the patient is being registered for treatment and email the completed registration 
checklist to the CRO at NCI Central Registration Office  ncicentralregistration-
l@mail.nih.gov. After confirmation of eligibility at Central Registration Office, CRO staff will 
call pharmacy to advise them of the acceptance of the patient on the protocol prior to the release 
of any investigational agents. Verification of Registration will be forwarded electronically via e-
mail to the research team. A recorder is available during non-working hours. 
Subjects that do not meet screening criteria should be removed from the study following the 
procedure in section 3.6.3.

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Version Date: June 13, 2018
18
2.3.1
Treatment Assignment and Randomization/Stratification Procedures for registration
purposes only
Cohorts
Number
Name
Description
1
Cohort 1
Patients with HPV 16 positive vulvar HSIL enrolled to 
determine the MTD
2
Cohort 2
Patients with HPV 16 positive vulvar HSIL enrolled 
after MTD is determined
Arms
Number
Name
Description
1
Arm 1
Patients will undergo leukapheresis, then treatment with 
E6 TCR cells (at escalating doses) + aldesleukin
2
Arm 2
Patients will undergo leukapheresis, then treatment with 
E6 TCR cells (at the MTD) + aldesleukin
Stratifications
Not applicable to this protocol.
Randomization and Arm Assignment
This is not a randomized study.  Patients in Cohort 1 will be directly assigned to Arm 1. Patients 
in Cohort 2 will be directly assigned to Arm 2.
2.4
BASELINE PROCEDURES
a. Baseline photographs and optional biopsies of HSIL lesions to be performed under local 
anesthetic in the clinic within 8 weeks prior to therapy. Patients will be required to sign a 
separate consent for all biopsies conducted during the study.  Refer to Section 5.4 for 
guidelines for handling specimens. 
b. Peripheral blood lymphocytes (PBLs), plasma, and serum collected at time of baseline 
biopsy. As noted in 5.1.3, Biopsies will not be performed if they are not technically 
feasible, if they will interfere with response assessment, or if a patient declines consent.  
c.
Replication Competent Retrovirus (RCR) testing.
d. Leukapheresis as described in section 3.1.2.

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19
3
STUDY IMPLEMENTATION
3.1
STUDY DESIGN
This is a phase I clinical trial with a 3+3 dose escalation design.

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Version Date: June 13, 2018
20
3.1.1
Protocol schema
1 month
Enrollment
Leukapheresis & Biopsy
E6 TCR Cells
+ aldesleukin
Safety 
Safety Visit, Leukapheresis & Biopsy (10-14 days)
1 week
1 week
Response Assessment 
& Biopsy (27-34 days)
Response Assessment               (2 months)
Response Assessment (3 months)
2 weeks
1 month
NR
Standard of 
Care 
CR or PR
Response Assessment q 1 
month x 3 (4, 5 and 6 month)
PR
CR
Standard of 
Care Treatment
Followup every 3 months for 2 
visits, then every 6 months for 2 
visits (2 years of followup total)

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21
3.1.2
Leukapheresis
The patient will undergo a 10-15 liter leukapheresis (generally, 12 liters will be processed to target 
yield of 6-10 x109 lymphocytes) in the Department of Transfusion Medicine (DTM) Dowling 
Apheresis Clinic according to DTM standard operating procedures.  The procedure requires dual 
venous access, and takes approximately 3-4 hours to complete.  A central line will be placed if 
peripheral venous access is not sufficient. The leukapheresis collection will be obtained at least 21 
days prior to the cell infusion. Leukapheresis material that is not required for clinical use will be 
retained and cryopreserved in 10 vials at 100x106 cells per vial with remaining cells stored at 
300x106 cells per vial for research and banked on protocol 16-C-0061 (ETIB Tissue Procurement 
Protocol).
3.1.3
E6 TCR T cell preparation
After cells are obtained by apheresis, further cell processing to generate E6 TCR cells will occur 
in the DTM according to standard operating procedures and the E6 TCR investigational new drug 
application. E6 TCR cells can be produced in approximately 21 to 27 days. Cell products may be 
cryopreserved during production to accommodate patient treatment schedules. Either freshly-
collected cells or cryopreserved cells can be used to initiate the cell-preparation process.  
Peripheral blood mononuclear cells (PBMC) will be isolated. Sufficient cells for three complete 
cell productions (2-3 vials at 3-4.5 x 109 cells/vial) will be retained in the DTM; the remaining 
cells will first be frozen in 10 vials at 100 x 106 cells per vial with excess frozen at 300 x 106
cells/vial. Cells will be frozen in the DTM and then transferred to ETIB the following 
day. Contacts in the ETIB preclinical core are Fran Hakim and Jeremy Rose (301-594-5339).  
Before infusion, the percentage of T cells expressing the E6 TCR will be determined by flow 
cytometry. Successful TCR gene transfer will be defined as greater than 20% TCR positive cells.
In addition to flow cytometry, further testing of the cells will take place prior to infusion to evaluate 
for microbial contamination, replication-competent retroviruses, and viability.  Details of this 
testing can be found in the appropriate DTM SOPs. Any remaining pretreatment PBMC collected 
on this protocol and cryopreserved as described in Section 3.1.2 will be transferred from the 
Department of Transfusion Medicine to the Principal Investigator of this protocol for storage in 
the ETIB preclinical core and possible use in research and banked according to protocol 16-C-
0061 (ETIB Tissue Procurement Protocol). 
3.1.4
Treatment Phase
E6 TCR T cells will be infused while the patient is admitted to the hospital on treatment day 0. A
central line catheter may be placed for cell infusion. The day after cell infusion, patients will 
receive aldesleukin 720,000 IU/kg IV q 12 hours x up to 2 doses. The second dose of aldesleukin 
will not be given if a patient develops any of the following: 1) hypotension or oliguria that 
require fluid boluses, 2) mental status changes, 3) hypoxia that requires supplemental oxygen, 4) 
diarrhea or nausea that are not controlled with symptomatic therapy. The second dose of 
aldesleukin may also be held for any safety concern at the discretion of the treating physician, 
and will not be given if a grade 3 or higher AE occurs following the first dose. 
The patient will remain an inpatient for aldesleukin administration. Patients also will be observed 
overnight as inpatients following aldesleukin administration, and will not be discharged until 
cleared by the treating physician. Patients will receive one course of treatment.  

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22
3.1.5
Dose Limiting Toxicities (DLTs)
All treatment related Grade 3 and greater AEs occurring within 30 days of the cell infusion will 
be considered DLTs with the exception of the following expected transient effects of aldesleukin.
x
Grade 3 fever or chills responsive to symptomatic treatment WKDWUHVROYHWRJUDGH
in 48 hours.
x
Grade 3 hypotension or oliguria responsive to /RI,9IOXLGEROXVHVLQKRXUV
WKDWUHVROYHVWRJUDGHLQKRXUV.
x
Grade 3 dyspnea/hypoxia that improves to grade 2 or less with supplemental 
oxygen DQGUHVROYHVWRJUDGHZLWKRXWVXSSOHPHQWDOR[\JHQLQKRXUV
x
Grade 3 creatinine or electrolyte abnormalities WKDWUHVROYHWRJUDGHLQKRXUV
x
Grade 3 or higher white blood cell count decreased and/or lymphocyte count 
decreased WKDWUHVROYHVWRJUDGHLQKRXUV
Further outline of expected AEs with aldesleukin and their management is included in 
Appendix B and Appendix C.
3.1.6
Dose Escalation
The protocol will follow a 3+3 phase 1 dose escalation design. Three patients will be treated 
initially on dose level -1. If 2 of these 3 patients experience DLTs, the study will be stopped. If 
none of the 3 patients experiences a DLT, enrollment can start on dose level 1. If 1 of the first 3
patients experiences a DLT on dose level -1, 3 more patients will be treated at that dose level.  If 
a second patient of the 6 has a DLT, the study will be stopped; otherwise, accrual can proceed to 
dose level 1. On dose level 1, if 2 patients have DLTs at any time during the treatment of 6 
patients, then treatment on dose level 1 will be stopped, and 3 additional patients will be accrued 
at dose level -1 to reach a total of 6, if only 3 were previously treated. The maximum tolerated 
dose is the highest dose at which a maximum of 1 of 6 patients has a DLT.
There will be a minimum of a 7-day delay in treatment between the patients and a minimum of a 
14-day delay in treatment between cohorts.
The total number of E6 TCR T cells transferred for each cohort will be:
x
Dose level -1 0.7 x 1010 transduced T cells
x
Dose level 1
0.7 x 1011 transduced T cells
The cell dose administered will be in a range of +/- 30% of the target dose above. The frequency 
of transduced T cells will be determined by flow cytometry with anti-mouse TCR beta-chain 
antibody.  The number of transduced cells to be administered will be determined by multiplying 
the frequency of transduced T cells by the total number of viable T cells. If fewer than the target 
number of cells are generated the patient will be treated but toxicity data from the patient will not 
be used in determining the MTD, and that patient will be replaced in the protocol enrollment.
3.1.7
Safety Protocol Stopping Rules
The study may be halted if any of the following safety conditions are met:

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23
1.
If one or more deaths occur, we will immediately stop accrual and promptly discuss this 
with the NCI IRB and the FDA. 
2.
If two or more patients develop a Grade 3 or greater toxicity not attributable to the 
aldesleukin (or circumstances unrelated to the study) that does not resolve to Grade 2
within 24 hours.
3.
If two patients develop Grade 3 or greater Immune Related Adverse Event (IRAE) related 
to aldesleukin that cannot be resolved to less than or equal to a Grade 2 within 10 days.
4.
If 2 DLTs occur on dose level 1. 
3.2
DRUG ADMINISTRATION
Inpatient treatment will be according to the following schedule. Cell infusions may be slowed or 
delayed as medically indicated. Administration of diuretics, hydration, and electrolyte 
monitoring and replacement will be performed as clinically indicated.
Day 0 
E6 TCR cells
The E6 TCR cells will be delivered to the patient care unit by an authorized staff 
member.  Prior to infusion, the cell product identity label is double-checked by two 
authorized staff (MD or RN) and an identification of the product and documentation of 
administration are entered in the patient’s chart (as is done for blood banking protocols).
E6 TCR T cells will be administered intravenously over 20 to 30 minutes via non-
filtered tubing, gently agitating the bag during infusion to prevent cell clumping. A
peripheral IV may be used if access is adequate (in general, two 18 gauge or larger IVs 
should be established). A PICC line or other central line may be used if peripheral access 
is not adequate.
Prophylactic antibiotics
Prophylactic antibiotics will be administered starting the morning of cell infusion and 
continue until 24 hours after the last dose of aldesleukin per PI discretion. Oxacillin,
clindamycin, or other antibiotic with equivalent gram positive coverage may be chosen 
based on patient’s history of allergies and physician discretion. 
Day 1
Beginning in the morning of day 1, Aldesleukin will be given 720,000 IU/kg IV q12 
hours for up to 2 doses. Infusion will take place over 15 minutes. The second dose of 
aldesleukin will be given at the treating physician’s discretion and will not be given if a 
grade 3 or higher AE occurs following the first dose. 

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Version Date: June 13, 2018
24
3.3
STUDY CALENDAR
Procedure
Screening/
Baseline 2
Day
0
Day
1
Day
2
Day
3
Daily
until d/c
Follow-up Period
1 wk
(Days 5-8)
2 wk
(Days 10-14)
Monthly f/u 8, 9
(Month 1 is also
End of Treatment
Visit)
Annual
Followup
History, physical exam, and review of systems1
X
X1
X1
X1
X1
X1
X
X
X
Vital signs
X
X
X
X
X
X
X
X
X
Concomitant medications
X
X
X
X
X
X
X
X
X
ECOG performance status
X
X
X
X
Chest x-ray
X
EKG/Cardiac stress test (as per guidelines in
Section 2.2)
X
Screening labs: HIV, HBV serology, HCV
serology, thyroid panel, PT/PTT, Beta-HCG
pregnancy test, HLA-A typing for HLA-A*02:01
allele positivity
X
HPV-16 verification and pathology review for
confirmation of diagnosis3
X
Acute care panel, hepatic panel, mineral panel,
LD, total protein, CK, uric acid, CBC with
differential and TBNK
X
X
X
X

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25
Procedure
Screening/
Baseline 2
Day
0
Day
1
Day
2
Day
3
Daily
until d/c
Follow-up Period
1 wk
(Days 5-8)
2 wk
(Days 10-14)
Monthly f/u 8, 9
(Month 1 is also
End of Treatment
Visit)
Annual
Followup
HSIL measurements, vulvar biopsy, and
photographs4
X
X
X
Leukapheresis
X
X
Replication Competent Retrovirus (RCR) testing5
X
X5
Research samples: peripheral blood lymphocytes,
plasma, and serum6
X
X
X
X6
X
X
X
E6 TCR T cells
X
Aldesleukin
X
Prophylactic antibiotics7
X7
X7
X7
Response Evaluation
X
HSIL and patient status update11
X
Adverse Events
X
X
X
X
X
X
X
X
1 Patients will have a full history and physical at screening, on admission to the hospital, and on follow-up visits. While inpatient, patients will have a daily
review of systems and physical exam.

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2 Exact timeline is indicated in Section 2.2
3 Vulvar HSIL must be HPV-16+ by PCR, RNA, or in situ hybridization test from a CLIA certified laboratory prior
to treatment.
4 Vulvar biopsies will be performed as a punch biopsy under local anesthetic in the outpatient clinic within 8 weeks
prior to starting therapy, at 2-week follow-up (day 10-14), and at 1-month follow-up (day 27-34) for all patients. An
additional biopsy will be obtained at the time a patient comes off-treatment (e.g. at time of progression, at time of
complete response, or at 6-month visit for patients with continuing partial response). Biopsies will be performed
only if technically feasible, if they do not interfere with response assessment, and if the patient provided consent.
The investigators may elect not to perform biopsies at their discretion (i.e. safety concerns, patient concerns, etc.).
HSIL photographs will occur at each follow up visit.
5 Patients’ blood samples will be obtained and undergo analysis for detection of replication competent retroviruses
(RCR) by PCR prior to cell infusion and at approximately 3, 6, and 12 months post cell administration. Blood
samples will be archived annually thereafter if all previous testing has been negative with a brief clinical history.
The ETIB preclinical core staff will cryopreserve 2 vials of cells if possible. If less than 10 million cells are
available, only 1 vial of cells will be cryopreserved.
6 Research samples will be collected at screening, pre-treatment (within four weeks prior to cell infusion);
posttreatment on post-therapy day 1, post-therapy day 3, thereafter any MWF while still an inpatient for one week,
and thereafter weekly while still inpatient; and post-treatment at each research visit as described in section 3.5. For
each time point, six 8mL CPT tubes will be drawn (5 for PBMC and 1 for plasma) and one 8mL SST tube (for sera).
Processing and storage are as described in section 5.
7 Prophylactic antibiotics will be administered starting the morning of cell infusion and continue until 24 hours after
the last dose of aldesleukin per PI discretion. If patients are discharged prior to completing antibiotics they will be
given a prescription to complete the course at home. Oxacillin, clindamycin, or other antibiotic with equivalent gram
positive coverage may be chosen based on patient’s history of allergies and physician discretion.
8 All patients follow- up 1, 2, and 3 months after cell infusion. Patients with no response at the 3-month evaluation
will come off-treatment and receive standard of care therapy. Patients with a partial or complete response at the 3-
month evaluation will continue follow up at 4, 5, and 6 months for safety, response, and research evaluations. After
the 6-month response assessment patients with partial response will receive standard of care therapy and patients
with a complete response will continue to have follow-up every 3 months (+/- 30 days) for 2 visits, then every 6
months (+/- 30 days) for 2 visits (2 years of follow up total). Patients with disease progression at any time will come
off treatment and receive standard of care therapy either at the NIH, with their primary physician, or another
institution. For follow up visits, if unwilling or unable to travel to the NIH Clinical Center, patients will be
contacted by telephone regarding their status, and may be asked to send labs and physical exam reports to fulfill
follow up visit requirements. Adverse events associated with standard of care therapy will not be recorded or
reported on this study.
9As indicated in 3.5 (+/- 14 days)
10As indicated in section 10.4DOOVXEMHFWVDJHZLOOEHRIIHUHGWKHRSSRUWXQLW\WRFRPSOHWHDQ1,+DGYDQFHG
directives form.  This should be done preferably at baseline but can be done at any time during the study as long as 
the capacity to do so is retained.  The completion of the form is strongly recommended, but is not required.
11As indicated in section 3.5.2, subjects will be contacted to collect information annually for five years after the date 
of cell infusion.  This follow up can be conducted via telephone.
3.4
ON-STUDY EVALUATIONS
3.4.1
Prior to starting cell infusion:
Within 14 days prior to therapy, patients will have a complete blood count, serum
chemistries performed including electrolytes, BUN, creatinine, pregnancy test and liver function

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27
tests. If any results are beyond the criteria established for eligibility, the patient will not proceed
until the abnormalities can be resolved.
x
Pre-treatment biopsy within 8 weeks prior to therapy
x
Blood samples for analysis for detection of Replication Competent Retrovirus (RCR) 
by PCR as described in Section 5.3.
3.4.2
After Cell Infusion:
x
Vital signs will be monitored hourly (+/- 15 minutes) for 4 hours and then routinely 
(every 4-6 hours) unless otherwise clinically indicated for the duration of the hospital 
stay
3.4.3
During Hospitalization:
Every 1-2 days.
x
Daily review of systems and physical exam
x
Other tests will be performed as clinically indicated.
3.5
POST TREATMENT EVALUATION (FOLLOW-UP)
Follow-up visits will be done according to the following schedule. 
x
1 week safety and research visit (days 5-8)
x
2 week safety and research visit (days 10-14)
x
1 month safety, research, and response visit (+/- 7 days)
x
2 month safety, research, and response visit (+/- 7 days)
x
3 month safety, research, and response visit (+/- 7 days)
x
4 month safety, research, and response visit (+/- 7 days)
x
5 month safety, research, and response visit (+/- 7 days)
x
6 month safety, research, and response visit (+/- 7 days)
x
As clinically indicated subsequently.
Note: Patients may be seen more frequently as clinically indicated
3.5.1
Evaluation:
x
Patients will follow up 1 and 2 weeks after cell infusion for safety assessment and 
research studies as indicated on the Study Calendar. Response will not be measured 
at these visits. 
x
Patients will follow up 1, 2, and 3 months after cell infusion for safety assessment,
research studies, and response evaluations.
x
Patients with no response at the 3 month evaluation will come off-treatment and 
receive standard of care therapy.
x
Patients with a partial or complete response at the 3-month evaluation will follow up 
at 4, 5, and 6 months for safety, response, and research evaluations.
x
After the 6-month response assessment patients with partial response will receive 
standard of care therapy. Patients with a complete response will thereafter have 
follow-up every 3 months (+/- 30 days) for 2 visits, then every 6 months (+/- 30 days) 
for 2 visits (2 years of follow up total).
x
Patients with disease progression at any time will come off treatment and receive 
standard of care therapy. 

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x
Patients who come off treatment will follow up as required for gene transfer studies
or as clinically indicated.
3.5.2
At each Response Evaluation, patients will undergo the following:
x
Physical examination
x
Chem 20 equivalent: (Sodium (Na), Potassium (K), Chloride (Cl), Total CO2 
(Bicarbonate), Creatinine, Glucose, Urea nitrogen (BUN), Albumin, Calcium total, 
Magnesium total (Mg), Inorganic Phosphorus, Alkaline Phosphatase, ALT/GPT, 
AST/GOT, Total Bilirubin, Direct Bilirubin, LD, Total Protein, Total CK, Uric Acid).
x
Complete Blood Count with differential and TBNK
x
Toxicity assessment, including a review of systems.
x
Serum, plasma, and peripheral blood lymphocytes.
x
HSIL measurements and photographs will be taken for documentation of response. 
x
Optional biopsies for research purposes will be obtained at baseline (within 8 weeks 
prior to starting treatment); at 2 week (day 10-14) safety and research visit; at 1 
month (day 27 to 34) safety, response, and research visit; and at the time the patient 
comes off-treatment (i.e. attains a complete response, experiences progression, or 
reaches the 6-month time point without a complete response). Biopsies will be 
performed only if technically feasible, if they do not interfere with response 
assessment, and if the patient provides consent. The investigators may elect not to 
perform biopsies at their discretion (i.e. safety concerns, patient concerns, etc.).
Biopsies for clinical concern will be performed as clinically indicated. Biopsies will 
be processed as per Section 5.1.3.
x
An approximately 5 liter leukapheresis will be performed at the 2 week (day 10-14)
follow up visit. If the patient is unable to undergo apheresis, approximately 96 ml of 
blood may be obtained. Subsequently, approximately 60 ml of blood will be obtained 
at each response visit (see Study Calendar for sampling details). Peripheral blood 
mononuclear cells will be cryopreserved for immunologic testing and will be banked 
under protocol 16-C-0061 (ETIB Tissue Procurement Protocol).  PBMC from 
apheresis will be stored in 10 vials at 1x108 cells per vial with remaining cells stored 
at 3x108 cells per vial. 
x
Cervical testing for high-risk HPV if the screening assessment was positive for HPV-
16.
x
The following information will be collected annually for a total of five years after the 
date of cell infusion: 1) procedures or treatments for vulvar HSIL, 2) procedures or 
treatments for cervical, vaginal, or anal intraepithelial neoplasia, 3) occurrence of 
vulvar, cervical, or anal cancer.
3.5.3
Patients who are unable or unwilling to return for follow up evaluations
They will be followed via phone or e-mail contact. Patients may be asked to send laboratory, 
imaging and physician exam reports performed by their treating physician.
3.5.4
Long-term Follow-Up:
Long-term follow-up of patients receiving gene transfer is required by the FDA 
and must continue even after the patient comes off the study.  Long-term follow-up

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29
will be done under a different protocol, 15-C-0141. After the patient comes off 
study, health status data will be obtained from surviving patients via telephone 
contact or mailed questionnaires for a total of 15 years after cell infusion. 
3.6
CRITERIA FOR REMOVAL FROM PROTOCOL THERAPY AND OFF STUDY CRITERIA
Prior to removal from study, effort must be made to have all subjects complete a safety visit 
approximately 30 days following the last dose of study therapy. Since patients will be 
completing a one-month follow up visit, the end of treatment visit will be the 30 day followup 
visit as indicated on the Study Calendar in Section 3.3
3.6.1
Criteria for removal from protocol therapy
x
Completion of protocol therapy.
x
Grade 3 or greater autoimmunity that involves vital organs (heart, kidneys, brain, eye, 
liver, colon, adrenal gland, lungs). 
x
Positive pregnancy test
x
If a patient experiences grade 3 or 4 toxicity due to cell infusion (reaction to cellular 
product or infusion reaction), the infusion will be stopped and the patient will receive 
no further treatment (i.e. aldesleukin).
x
Progression of disease
x
Participant requests to be withdrawn from active therapy
x
Investigator discretion
3.6.2
Off-Study Criteria
x
Screen failure
x
Completed study follow-up period
x
Inability to generate a cell product. A second attempt may be made to generate a cell 
product from the patient.   If the second attempt fails, that patient will be removed 
from the study and replaced with another patient.  
x
Participant requests to be withdrawn from study
x
There is significant noncompliance
x
General or specific changes in the patient’s condition render continued participation 
of the patient unacceptable for further follow up in the judgment of the investigator.
x
Death
3.6.3
Off Protocol Therapy and Off-Study Procedure
Authorized staff must notify Central Registration Office (CRO) when a subject is taken off 
protocol therapy and when a subject is taken off study. A Participant Status Update Form from 
the web site (http://home.ccr.cancer.gov/intra/eligibility/welcome.htm) main page must be 
completed and sent via encrypted email to: NCI Central Registration Office 
ncicentralregistration-l@mail.nih.gov .

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30
4
CONCOMITANT MEDICATIONS/MEASURES
4.1
STEROIDS
Patients needing systemic steroid therapy may not participate in this study.  
4.1.1
Blood Product Support
Patients are unlikely to require blood product support as a side effect of therapy. However, 
any patient on study may receive platelets and packed red blood cells (PRBC’s) in clinically 
needed.  As a general guideline, patients may be transfused for: 
x
Hemoglobin < 8 gm/dl
x
Platelets < 10,000/mm3
All blood products will be irradiated and leukocyte filters will be utilized to decrease 
sensitization to transfused WBC’s.
4.2
OTHER CONCOMITANT MEDICATIONS/MEASURES
Concomitant medications to control side effects of therapy may be given.  Meperidine 
(25-50 mg) will be given intravenously if severe chilling develops.  Other supportive therapy 
will be given as required and may include acetaminophen (650 mg q4h), indomethacin (50-75
mg q8h) and ranitidine (150 mg g12h). If patients require steroid therapy they will be taken off 
treatment.  Patients who require transfusions will receive irradiated blood products.  Ondansetron 
will be administered per standard dosing instructions as needed for nausea and vomiting.  
Additional antiemetics will be administered as needed for nausea and vomiting uncontrolled by 
ondansetron.  Antibiotic coverage will be provided at the discretion of the investigator.
5
BIOSPECIMEN COLLECTION
5.1
CORRELATIVE STUDIES FOR RESEARCH
The amount of blood that may be drawn from adult patients for research purposes shall not 
exceed 10.5 mL/kg or 550mL; whichever is smaller, over any 8-week period.  
5.1.1
Pre cell infusion evaluations
x
The following samples will be drawn for research and sent to the ETIB preclinical core
within 4 weeks prior to cell infusion. Send to Dr. Fran Hakim, Pre-Clinical Core lab;
Attention Jeremy Rose, Bldg 10, room 12C216 contact phone: 301-594-5339.
o
6 CPT tubes (8mL each). One CPT tube per time point will be used to collect 
4mL of plasma, which will be frozen in 4mL vials. PBMC from the remainder of 
the CPT tubes will be frozen in aliquots of 10 x 106 cells/vial
o
1 SST tube (8mL) for sera. Serum will be processed in the ETIB Pre-Clinical 
Core and aliquoted into vials of 0.5-1 mL each. 
x
During screening, one 10 mL tube of blood will be collected from each patient in a Cell-
Free DNA BCT tube.  Specimens may be drawn at the patient’s local doctor’s office or 
laboratory and FedExed to the assigned research nurse.  Specimens will be transported by 
the assigned research nurse to Dr. Christian Hinrichs’ lab for sample labeling.  Contact 
Stacey Doran, Bldg 10, room 4B-04. HPV-16 in peripheral blood via ddpCR assay
samples will be coded with a patient identification number, and linked to samples by that 
identification number in LabMatrix. Access to personally identifiable information (PII) is

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31
limited to the PI and study personnel who interact directly with the patient and their 
samples.  Once labeled, samples will be sent to the laboratory of Liang Cao, Genetics 
Branch, CCR for HPV-16 ddpCR assay testing.  See Appendix E for collection and 
processing instructions, which will be sent to the patient.
5.1.2
Post cell infusion evaluations
x
The following samples will be drawn for research and sent to the ETIB preclinical core
While the patient is inpatient, samples will be drawn on post-treatment day 1, post-
treatment day 3, thereafter every Monday/Wednesday/Friday x5 days, and then weekly
until discharge. Samples will also be collected at each outpatient follow-up visit. Send to
Dr. Fran Hakim, Pre-Clinical Core lab; Attention Jeremy Rose, Bldg 10, room 12C216
contact phone: 301-594-5339.
o
6 CPT tubes (8mL each). One CPT tube per time point will be used to collect 
4mL of plasma, which will be frozen in 4mL vials. PBMC from the remainder of 
the CPT tubes will be frozen in aliquots of 10 x 106 cells/vial
o
1 SST tube (8mL) for sera. Serum will be processed in the ETIB Pre-Clinical 
Core and aliquoted into vials of 0.5-1 mL each. 
Aliquots of serum samples will be provided to the laboratory of Liang Cao, Genetics 
Branch, CCR for testing of plasma or serum for HPV DNA.
5.1.3
Optional vulvar Biopsies
x
Biopsies of vulvar lesions for clinical purposes will be obtained within 8 weeks prior to 
starting treatment if clinically indicated to rule out invasive cancer. Biopsies will be 
performed in the outpatient clinic under local anesthetic. 
x
Biopsies for research purposes will be obtained at baseline (within 8 weeks of starting 
treatment), 10 to 14 days after treatment, 27 to 34 days after treatment, and at the time the 
patient comes off-treatment (i.e. attains a complete response, experiences progression, or 
reaches the 6-month time point without a complete response). Clinically indicated 
biopsies may be obtained at any time by PI discretion if there is concern for invasive 
cancer.
x
Biopsies for research will be performed with a standard punch biopsy instrument of up to 
4 mm diameter. At each time point, up to 3 biopsies of HSIL lesions and up to 2 biopsies 
of adjacent normal tissue will be performed. A formalin-fixed paraffin embedded 
specimen will be transported to the NCI Laboratory of Pathology for review and 
archiving. The contact will be Stacey Doran, Bldg 10, Room 4B04, contact 301-491-
6957. Specimens will then be transported to the ETIB preclinical core for further 
processing and storage. They will be delivered to Dr. Fran Hakim, Pre-Clinical Core lab; 
Attention Jeremy Rose, Bldg 10, room 12C216 contact phone: 301-594-5339.
x
Processing of biopsies will depend upon the amount of tissue obtained. Examples of 
processing may include OTC blocks, formalin-fixed paraffin embedding, snap freezing, 
placing in RNAlater or other required laboratory processing technique. 
x
Biopsies will not be performed if they are not technically feasible, if they will interfere 
with response assessment, or if a patient declines consent.

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32
x
Tissue samples will be coded and deidentified prior to transfer to the laboratory of Liang 
Cao, Genetics Branch, CCR for quantification of HPV DNA.
x
Some of these samples will be archived and analyzed under another protocol 16-C-0061
(ETIB Tissue Procurement Protocol) if the subject is also enrolled on that study.
5.1.4
Immunological Testing
x
Leukapheresis will be performed prior to treatment and 10-14 days after treatment.  
x
Processing of leukapheresis will be in the Department of Transfusion Medicine (DTM). 
Cell product will be frozen in 10 vials at concentration 100 x 106 cells/mL and additional 
vials at 300 x 106 cells/mL. Cells will be stored in a -80 degrees Celsius freezer in DTM 
overnight and transferred to the ETIB Preclinical Core the following day. Samples will be 
sent to Dr. Fran Hakim, Pre-Clinical Core lab; Attention Jeremy Rose, Bldg 10, room 
12C216. Contact phone: 301-594-5339.
x
At other time points, peripheral blood lymphocytes (PBL) and plasma will be obtained 
from whole blood by purification using centrifugation. These samples will be transferred 
directly to the ETIB Pre-Clinical Core lab for processing. Plasma will be frozen in 4mL 
vials. PBL will be frozen in aliquots of 10 x 106 cells/vial.
x
A variety of tests including but not limited to the following will be used to evaluate 
immunologic correlates: specific lysis and cytokine release, intracellular FACS of 
cytokine production, ELISPOT assays, and lymphocyte subset analysis. 
x
Samples of all infused cell products will be cryopreserved, and extensive retrospective 
analysis of infused cell phenotype and function will be performed to attempt to find in 
vitro characteristics of the infused cells which correlate with in vivo activity. 
x
Peripheral blood lymphocytes (PBL) may be tested by cytolysis assays, cytokine release, 
limiting dilution analysis, and other experimental studies.  Immunological monitoring 
will consist of quantifying T cells reactive with tumor cells using established techniques 
such as intracellular FACS, cytokine release assays, and ELISPOT assays.  
Immunological assays will be standardized by the inclusion of 1) pre-infusion PBMC and 
2) an aliquot of the T cells cryopreserved at the time of infusion. 
x
Blood and tissue specimens collected in the course of this research project may be banked 
and used in the future to investigate new scientific questions related to this study and 
protocol 16-C-0061 (ETIB Tissue Procurement Protocol).
5.2
SUMMARY OF SAMPLE COLLECTION
Test/assay
Volume blood 
(approx)
Type of 
tube
Collection point 
(+/- 48hrs)
Location of specimen analysis
Whole Blood
10 mL
Cell-free 
DNA BCT 
Screening
Processed in Christian Hinrichs Lab, 
then transported to Dr. Liang Cao’s 
lab for analysis
Plasma/PBMC
48 mL
CPT 
<4 weeks prior to cell 
infusion
Pre-Clinical core lab (Dr. Fran 
Hakim’s lab)

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33
Test/assay
Volume blood 
(approx)
Type of 
tube
Collection point 
(+/- 48hrs)
Location of specimen analysis
Serum
8 mL
SST
<4 weeks prior to cell 
infusion
Pre-Clinical core lab (Dr. Fran 
Hakim’s lab)
Plasma/PBMC
48 mL
CPT
Post cell infusion day 1, 
3, every Mon/Wed/Fri x 
5 days, weekly until 
discharge
Pre-Clinical core lab (Dr. Fran 
Hakim’s lab)
Serum
8mL
SST
Post treatment day 1, 3, 
every Mon/Wed/Fri x 5 
days, weekly until 
discharge
Pre-Clinical core lab (Dr. Fran 
Hakim’s lab)
Optional Vulvar 
Biopsy
N/A
N/A
< 8 weeks prior to 
treatment, 10-14 days 
after treatment, 27-34
days after treatment, and 
when patient comes off 
treatment.
NCI Laboratory of Pathology (for 
review and archiving)
Processed in Christian Hinrichs Lab, 
then transported to Pre-Clinical core 
lab (Dr. Fran Hakim’s lab)
Dr. Liang Cao’s lab
5.3
GENE-THERAPY-SPECIFIC FOLLOW-UP
x
Patients will be enrolled on the Long Term Followup Gene Therapy Study (Protocol 15-
C-0141) in order to collect information to meet current FDA requirements. All followup
as required by the FDA will occur via the 15-C-0141 protocol.
5.4
SAMPLE STORAGE, TRACKING AND DISPOSITION
5.4.1
Storage/Tracking in the Preclinical Development and Clinical Monitoring Facility
(PDCMF)
x
Samples will be ordered in CRIS and tracked through the Clinical Trial Data 
Management System.  Should a CRIS screen not be available, the CRIS downtime 
procedures will be followed.  Samples will not be sent outside the NIH without IRB 
notification and an executed MTA. 
x
Patient blood and tissue samples, collected for the purpose of research under IRB 
approved protocols of the Experimental Transplantation and Immunology Branch
(ETIB), may be archived by the ETIB Preclinical Development and Clinical Monitoring 
Facility (PDCMF). All data associated with archived clinical research samples is entered 
into the ETIB PDCMF’s Microsoft Excel databases on frozen cells and serum. These 
databases are stored on the NCI group drive in the ETIB ‘PRECLINSERVICE’ folder. 
Access to this folder is limited to PDCMF staff and ETIB clinical staff, requiring 
individual login and password. All staff in the PDCMF laboratory receive annually 
updated NIH/CIT training and maintain standards of computer security.

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x
The data recorded for each sample includes the patient ID, trial name/protocol number, 
date drawn, treatment cycle/post-transplant time point, cell source (e.g. peripheral blood, 
marrow,) as well as box and freezer location. Patient demographics that correlate 
treatment outcomes and therapies with the samples can be obtained only through the 
NCI/ETIB clinical records. As of January 2007, all newly received samples receive a 
unique bar code number, which is included in the sample record in the PDCMF database. 
Only this bar code is recorded on the sample vial and the vials will not be traceable back 
to patients without authorized access to the PDCMF database. All non-coded samples 
previously archived will be stripped of identifiers prior to distribution for any use other 
than as a primary objective of the protocol under which they were collected.
x
Samples are stored in freezers.  All samples will be labeled solely with a bar code (which 
includes the date, and serially determined individual sample identifier). The key will be 
available to a restricted number of ETIB investigators and associate investigators on the 
protocol. Coded samples will be stored frozen at -20°, -80° or liquid nitrogen vapor phase 
to -180 C according to the stability requirements in a single location under the restricted 
control of the PDCM Facility of ETIB.
These freezers are located onsite at the Preclinical Service laboratory (12C216) (-85° 
freezer) or in ETIB common equipment space (CRC/3-3273). Access to samples from a 
protocol for research purposes will be by permission of the Principal Investigator of that 
protocol in order to be used (1) for research purposes associated with protocol objectives 
for which the samples were collected, or (2) for a new research activity following 
submission and IRB approval of a new protocol and consent, or (3) for use only as unlinked 
or coded samples under the OHSRP Exemption Form guidelines stipulating that the 
activity is exempt from IRB review. Unused samples must be returned to the PDCMF 
laboratory. Samples, and associated data, will be stored permanently unless the patient 
withdraws consent. If researchers have samples remaining once they have completed all 
studies associated with the protocol, they must be returned to the PDCMF laboratory.
5.4.3
Storage/Track in the Christian Hinrichs lab
Blood, plasma, serum, and tissue samples will be processed, labeled, and stored in the ETIB 
Pre-Clinical Core. Samples will be requested and transferred to Dr. Hinrichs’ lab as needed 
for analysis. The contact will be Stacey Doran, Bldg 10, Room 4B04, phone 301-491-6957.
Once received, the samples will be coded utilizing LabMatrix.  The bar-coded samples will 
be stored at -80oC or in liquid nitrogen until time of use. Any unused samples will be 
returned to the PDCMF laboratory at the completion of all studies associated with the 
protocol. 
Samples transferred to the Hinrichs laboratory will be barcoded and tracked with 
LabMatrix.
Laboratory research data will be stored on the NCI secure server in the Hinrichs 
laboratory folder with secure access by laboratory personnel only. Access to personally 
identifiable information (PII) is limited to the PI and study personnel who interact 
directly with the patient and their samples.

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5.4.4
Storage/Tracking in the Liang Cao lab
Serum samples will be processed in the ETIB Pre-Clinical Core and 1 ml aliquots will be 
transferred to Dr. Cao’s lab in batches for circulating tumor DNA analysis.  Samples will 
be sent to Dr. Liang Cao, Molecular Targets Core Lab of Genetics Branch; Attention Dr. 
Zhigang Kang, Bldg 37, room 6134. Contact phone: 301-443-2817.
Once received, the bar-coded samples will be stored at -80oC until the time of testing.  
Serum DNA will be isolated with an automated purification system by Promega Corp 
using a system-attached bar-code reader to track samples and DNA products.  The 
circulating tumor/HPV DNA will be quantified with a digital droplet PCR system from 
Bio-Rad to obtain precise quantification.
5.4.4
Protocol Completion/Sample Destruction
x
Once research objectives for the protocol are achieved, researchers can request access to 
remaining samples, providing they have both approval of the Principal Investigator of the 
original protocol under which the samples or data were collected and either an IRB 
approved protocol and patient consent or an OHSRP exemption indicating that the 
activity is exempt from IRB review.
x
The PDCMF staff will report to the Principal Investigators any destroyed samples, if 
samples become unsalvageable because of environmental factors (ex. broken freezer or 
lack of dry ice in a shipping container), lost in transit between facilities or misplaced by a 
researcher. 
x
The PI will report destroyed samples to the IRB if samples become unsalvageable 
because of environmental factors (ex. broken freezer or lack of dry ice in a shipping 
container) or if a patient withdraws consent. Samples will also be reported as lost if they 
are lost in transit between facilities or misplaced by a researcher.  Freezer problems, lost 
samples or other problems associated with samples will also be reported to the IRB, the 
NCI Clinical Director, and the office of the CCR, NCI.
6
DATA COLLECTION AND EVALUATION 
6.1
DATA COLLECTION
The PI will be responsible for overseeing entry of data into an in-house password protected 
electronic system (C3D) and ensuring data accuracy, consistency and timeliness. The principal 
investigator, associate investigators/research nurses and/or a contracted data manager will assist 
with the data management efforts. All data obtained during the conduct of the protocol will be 
kept in secure network drives or in approved alternative sites that comply with NIH security 
standards. Primary and final analyzed data will have identifiers so that research data can be 
attributed to an individual human subject participant.
The key for assignment of patient code identification numbers with the personal identifiers will 
be stored in a secure data base. This key will not be shared with other investigators. Investigators 
conducting the individual sample testing will only have access to coded identification numbers 
and coded patient information (i.e. treatment regimens, treatment responses, diagnoses, 
pathology information).
The date on which screening consent is signed is the same date informed consent is signed.

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36
All adverse events, including clinically significant abnormal findings on laboratory evaluations, 
regardless of severity, will be followed until return to baseline or stabilization of event. Patients 
will be followed for adverse events for at least 30 days after removal from study treatment or 
until off-study, whichever comes first.
End of study procedures: Data will be stored according to HHS, FDA regulations and NIH 
Intramural Records Retention Schedule as applicable.
Loss or destruction of data: Should we become aware that a major breach in our plan to protect 
subject confidentiality and trial data has occurred, the IRB will be notified. 
6.1.1
Exclusions to Routine Adverse Event Reporting:
Adverse events in patients who go on to standard of care therapy will not be collected or reported 
for this study.
6.2
DATA SHARING PLANS
6.2.1
Human Data Sharing Plan
What data will be shared?
I will share human data generated in this research for future research as follows:
x__ De-identified data in an NIH-funded or approved public repository.  
_x__ De-identified data in BTRIS (automatic for activities in the Clinical Center)
_x__ De-identified or identified data with approved outside collaborators under appropriate 
agreements.
How and where will the data be shared? 
Data will be shared through:
x__ An NIH-funded or approved public repository.  Insert name or names: ClinicalTrials.gov.
_x__ BTRIS (automatic for activities in the Clinical Center)
_x__ Approved outside collaborators under appropriate individual agreements.
_x__ Publication and/or public presentations.
When will the data be shared
_x__ Before publication.
_x__ At the time of publication or shortly thereafter.

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6.3
RESPONSE CRITERIA
6.3.1
Definitions
Evaluable for toxicity: All patients will be evaluable for toxicity from the time of E6 TCR T cell
infusion. Toxicity data will be collected for all patients.
Evaluable for objective response: Only those patients who have measurable disease present at
baseline, have received an E6 TCR T cell infusion, and have had their disease re-evaluated for
response will be considered evaluable for response. These patients will have their response
classified according to the definitions stated below.
6.3.2
Disease Parameters
Measurable disease: Measurable lesions are defined as those that can be accurately measured in
at least one dimension as > 5 mm. All measurements must be recorded in millimeters (or
decimal fractions of centimeters).
Non-measurable disease. All other lesions (or sites of disease), including small lesions (longest
diameter <5 mm), are considered non-measurable disease.
Target lesions. All measurable lesions will be identified as target lesions and recorded and
measured at baseline.
Non-target lesions. All other lesions (or sites of disease) will be identified as non-target lesions
and should also be recorded at baseline. Measurements of these lesions are not required, but the
presence, absence, or in rare cases unequivocal progression of each will be noted throughout
follow-up.
6.3.3
Methods for Evaluation of Measurable Disease
Lesions will be photographed, described in detail, and measured at each response evaluation. 
Measurements will be recorded in metric notation using a ruler or calipers. Baseline evaluations 
will be performed no more than 8 weeks before starting treatment.
The same method of assessment will be used to characterize each identified and reported lesion
at baseline and during follow-up. Lesions will be documented by digital color photography at
each response evaluation.
6.3.4
Response Criteria
6.3.4.1
Evaluation of Target Lesions
Complete Response (CR): Disappearance of all target lesions. No appearance of new lesions.
Partial Response (PR): A  50% decrease in the sum of the product of the longest perpendicular
diameters of target lesions, taking as reference the baseline measurements. No appearance of
new lesions. No increase of greater than 25% of any index lesion.
Progressive Disease (PD): A  25% increase in sum of the product of the longest perpendicular 
diameters of target lesions, taking as reference the smallest product on study (this includes the 
baseline product if that is the smallest on study).  In addition to the relative increase of 25%, the 
sum must also demonstrate an absolute increase of at least 5 mm.  (Note: the appearance of one 
or more new lesions is also considered progression).

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Non-CR/Non-PD: Neither sufficient decrease to qualify for PR nor sufficient increase to qualify 
for PD, taking as reference the smallest product of diameters while on study.
Note: In the event of lesion regression with a residual abnormality of unclear significance, a 
biopsy may be obtained. If HSIL is present in the pathology specimen the response will be 
considered less than complete. If HSIL is not present the response will be considered complete. 
6.3.4.2
Evaluation of Non-Target Lesions
Complete Response (CR): Disappearance of non-target lesions.
Progressive Disease (PD): Appearance of one or more new lesions and/or unequivocal
progression of existing non-target lesions. It must be representative of the overall non-target
disease rather than an increase in a single lesion.
Non-CR/Non-PD: Persistence of one or more non-target lesion(s).
6.3.4.3
Evaluation of Best Overall Response
The best overall response is the best response recorded from the start of the treatment until
disease progression/recurrence (taking as reference for progressive disease the smallest
measurements recorded since the treatment started).
For Patients with Measurable Disease (i.e., Target Disease)
Target
Lesions
Non-Target
Lesions
New
Lesions
Overall Response
CR
CR
No
CR
CR
Non-
CR/Non-PD
No
PR
CR
Not evaluated
No
PR
PR
Non-
CR/Non-
PD/not
evaluated
No
PR
Non-
CR/Non-
PD
Non-
CR/Non-
PD/not
evaluated
No
Non-CR/Non-PD

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39
PD
Any
Yes or No
PD
Any
PD*
Yes or No
PD
Any
Any
Yes
PD
*
In exceptional circumstances, unequivocal progression in non-target lesions may be
accepted as disease progression.
Note:
Patients with a global deterioration of health status requiring discontinuation of
treatment without objective evidence of disease progression at that time should be
reported as “symptomatic deterioration.” Every effort should be made to document the
objective progression even after discontinuation of treatment.
6.3.5
Duration of Response
Duration of overall response:  The duration of overall response is measured from the time 
measurement criteria are met for CR or PR (whichever is first recorded) until the first date that 
recurrent or progressive disease is objectively documented (taking as reference for progressive 
disease the smallest measurements recorded since the treatment started).
Duration of overall CR: The duration of overall CR is measured from the time measurement 
criteria are first met for CR until the first date that progressive disease is objectively documented.
Duration of overall non-CR/non-PD: Non-CR/non-PD disease is measured from the start of the 
treatment until the criteria for progression are met, taking as reference the smallest measurements 
recorded since the treatment started, including the baseline measurements. 
6.4
TOXICITY CRITERIA
Careful evaluation to ascertain the toxicity, immunologic effects and anti-HSIL efficacy of the 
treatment regimens will be performed. This study will utilize the CTCAE version 4.0 for toxicity 
and adverse event reporting.  A copy of the CTCAE version 4.0 can be downloaded from the 
website http://ctep.cancer.gov.  All appropriate treatment areas should have access to a copy of 
the CTCAE version 4.0. A copy of the CTCAE version 4.0 can be downloaded from the CTEP 
web site (http://ctep.cancer.gov/protocolDevelopment/electronic_applications/ctc.htm#ctc_40).
7
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN
7.1.1
Adverse Event
An adverse event is defined as any untoward medical occurrence in a human subject, including 
any abnormal sign (for example, abnormal physical exam or laboratory finding), symptom, or 
disease, temporally associated with the subject’s participation in research, whether or not 
considered related to the subject’s participation in the research.

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7.1.2
Suspected adverse reaction
Suspected adverse reaction means any adverse event for which there is a reasonable possibility 
that the drug caused the adverse event. For the purposes of IND safety reporting, ‘reasonable 
possibility’ means there is evidence to suggest a causal relationship between the drug and the 
adverse event. A suspected adverse reaction implies a lesser degree of certainty about causality 
than adverse reaction, which means any adverse event caused by a drug.
7.1.3
Unexpected adverse reaction
An adverse event or suspected adverse reaction is considered “unexpected” if it is not listed in 
the investigator brochure or is not listed at the specificity or severity that has been observed; or, 
if an investigator brochure is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current application. 
"Unexpected” also refers to adverse events or suspected adverse reactions that are mentioned in 
the investigator brochure as occurring with a class of drugs or as anticipated from the 
pharmacological properties of the drug, but are not specifically mentioned as occurring with the 
particular drug under investigation.
7.1.4
Serious
An Unanticipated Problem or Protocol Deviation is serious if it meets the definition of a Serious 
Adverse Event or if it compromises the safety, welfare or rights of subjects or others.
7.1.5
Serious Adverse Event
An adverse event or suspected adverse reaction is considered serious if in the view of the 
investigator or the sponsor, it results in any of the following:
x
Death
x
A life-threatening adverse drug experience
x
Inpatient hospitalization or prolongation of existing hospitalization
x
Persistent or significant incapacity or substantial disruption of the ability to conduct 
normal life functions
x
A congenital anomaly/birth defect.
x
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse drug event when, based upon 
appropriate medical judgment, they may jeopardize the patient or subject and may require 
medical or surgical intervention to prevent one of the outcomes listed in this definition.
7.1.6
Disability
A substantial disruption of a person’s ability to conduct normal life functions.
7.1.7
Life-threatening adverse drug experience
Any adverse event or suspected adverse reaction that places the patient or subject, in the view of 
the investigator or sponsor, at immediate risk of death from the reaction as it occurred, i.e., it 
does not include a reaction that had it occurred in a more severe form, might have caused death.
7.1.8
Protocol Deviation (NIH Definition)
Any change, divergence, or departure from the IRB-approved research protocol.  

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7.1.9
Non-compliance (NIH Definition)
The failure to comply with applicable NIH Human Research Protections Program (HRPP)
policies, IRB requirements, or regulatory requirements for the protection of human research 
subjects.
7.1.10
Unanticipated Problem
Any incident, experience, or outcome that:
x
Is unexpected in terms of nature, severity, or frequency in relation to 
(a) the research risks that are described in the IRB-approved research protocol and 
informed consent document; Investigator’s Brochure or other study documents, and
(b) the characteristics of the subject population being studied; AND
x
Is related or possibly related to participation in the research; AND
x
Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized.
7.2
NCI-IRB AND CLINICAL DIRECTOR REPORTING
7.2.1
NCI-IRB and NCI Clinical Director Expedited Reporting of Unanticipated Problems
and Deaths
The Protocol PI will report in the NIH Problem Form to the NCI-IRB and NCI Clinical Director:
x
All deaths, except deaths due to progressive disease
x
All Protocol Deviations
x
All Unanticipated Problems
x
All non-compliance
Reports must be received within 7 days of PI awareness via iRIS.
7.2.2
NCI-IRB Requirements for PI Reporting at Continuing Review
To ensure safety using this treatment, the NCI ETIB will review safety data on all protocols semi-
annually at the time of continuing review.  Data will be presented for both the recent 6 month 
period and for the entire length of time the protocol has been open.  The toxicity data for review 
will include all toxicities captured on the protocol and will include a table that describes all 
incidences of intubation including the duration of and reason for intubation.
The protocol PI will report to the NCI-IRB: 
1. A summary of all protocol deviations in a tabular format to include the date the deviation 
occurred, a brief description of the deviation and any corrective action.
2. A summary of any instances of non-compliance
3. A tabular summary of the following adverse events:

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x
All Grade 2 unexpected events that are possibly, probably or definitely related to the 
research; 
x
All Grade 3 and 4 events that are possibly, probably or definitely related to the research; 
x
All Grade 5 events regardless of attribution; 
x
All Serious Events regardless of attribution. 
NOTE: Grade 1 events are not required to be reported. 
7.2.3
NCI-IRB Reporting of IND Safety Reports
Only IND Safety Reports that meet the definition of an unanticipated problem will need to be 
reported to the NCI IRB.
7.3
IND SPONSOR REPORTING CRITERIA
During the first 30 days after the subject receives the investigational agent, an investigator must 
immediately report to the sponsor, using the mandatory MedWatch form 3500a or equivalent,
any serious adverse event, whether or not considered drug related, including those listed in the 
protocol or investigator brochure and must include an assessment of whether there is a 
reasonable possibility that the drug caused the event. For serious adverse events that occur more 
than 30 days after the last administration of the investigational agent, only report those that have 
an attribution of at least possibly related to the agent/intervention.
Required timing for reporting per the above guideline:
•
Deaths (except death due to progressive disease) must be reported via email within 
24 hours.  A complete report must be submitted within one business day. 
•
Other serious adverse events as well as deaths due to progressive disease must be 
reported within one business day
Events will be submitted to the Center for Cancer Research (CCR) at: CCRsafety@mail.nih.gov
and to the CCR PI and study coordinator.
7.3.1
Reporting Pregnancy/Maternal exposure
If a patient becomes pregnant during the course of the study, the study treatment should be 
discontinued immediately and the pregnancy reported to the Sponsor. The potential risk of 
exposure of the fetus to the investigational agent(s) or chemotherapy agents (s) should be 
documented in box B5 of the MedWatch form “Describe Event or Problem”.
Pregnancy itself is not regarded as an AE unless there is a suspicion that the study treatment 
under study may have interfered with the effectiveness of a contraceptive medication. However, 
as patients who become pregnant on study risk intrauterine exposure of the fetus to agents which 
may be teratogenic, the CCR is requesting that pregnancy should be reported in an expedited 
manner as Grade 3 “Pregnancy, puerperium and perinatal conditions - Other (pregnancy)”
under the Pregnancy, puerperium and perinatal conditions SOC. 
Congenital abnormalities or birth defects and spontaneous miscarriages should be reported and 
handled as SAEs. Elective abortions without complications should not be handled as AEs. The 

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outcome of all pregnancies (spontaneous miscarriage, elective termination, ectopic pregnancy, 
normal birth, or congenital abnormality) should be followed up and documented.
If any pregnancy occurs in the course of the study, then the investigator should inform the 
Sponsor within 1 day, i.e., immediately, but no later than 24 hours of when he or she becomes 
aware of it.
The designated Sponsor representative will work with the investigator to ensure that all relevant 
information is provided to the Sponsor within 1 to 5 calendar days for SAEs and within 30 days 
for all other pregnancies.
The same timelines apply when outcome information is available.
7.4
INSTITUTIONAL BIOSAFETY COMMITTEE (IBC) REPORTING CRITERIA
7.4.1
Serious Adverse Event Reports to IBC
The Principal Investigator (or delegate) will notify IBC of any unexpected fatal or life-
threatening experience associated with the use of E6TCR transduced PBL cells as soon as 
possible but in no event later than 7 calendar days of initial receipt of the information. Serious 
adverse events that are unexpected and associated with the use of the E6 TCR transduced PBL,
but are not fatal or life-threatening, much be reported to the NIH IBC as soon as possible, but not 
later than 15 calendar days after the investigator’s initial receipt of the information.  Adverse 
events may be reported by using the FDA Form 3500a.
7.4.2
Annual Reports to IBC
Within 60 days after the one-year anniversary of the date on which the IBC approved the initial 
protocol, and after each subsequent anniversary until the trial is completed, the Principal 
Investigator (or delegate) shall submit the information described below.  Alternatively, the IRB 
continuing review report can be sent to the IBC in lieu of a separate report.  Please include the 
IBC protocol number on the report.
7.4.2.1
Clinical Trial Information
A brief summary of the status of the trial in progress or completed during the previous year. The 
summary is required to include the following information: 
x
the title and purpose of the trial
x
clinical site
x
the Principal Investigator
x
clinical protocol identifiers; 
x
participant population (such as disease indication and general age group, e.g., adult or 
pediatric); 
x
the total number of participants planned for inclusion in the trial; the number entered into 
the trial to date whose participation in the trial was completed; and the number who 
dropped out of the trial with a brief description of the reasons
x
the status of the trial, e.g., open to accrual of subjects, closed but data collection ongoing, 
or fully completed, 

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x
if the trial has been completed, a brief description of any study results. 
7.4.2.2
Progress Report and Data Analysis
Information obtained during the previous year's clinical and non-clinical investigations, 
including: 
x
a narrative or tabular summary showing the most frequent and most serious adverse 
experiences by body system
x
a summary of all serious adverse events submitted during the past year
x
a summary of serious adverse events that were expected or considered to have causes not 
associated with the use of the gene transfer product such as disease progression or 
concurrent medications
x
if any deaths have occurred, the number of participants who died during participation in 
the investigation and causes of death
x
a brief description of any information obtained that is pertinent to an understanding of the 
gene transfer product’s actions, including, for example, information about dose-response, 
information from controlled trials, and information about bioavailability.
7.5
DATA AND SAFETY MONITORING PLAN
7.5.1
Principal Investigator/Research Team
The clinical research team will meet on a regular biweekly basis when patients are being actively 
treated on the trial to discuss each patient. Decisions about dose level enrollment and dose 
escalation if applicable will be made based on the toxicity data from prior patients. 
All data will be collected in a timely manner and reviewed by the principal investigator. Adverse 
events will be reported as required above. Any safety concerns, new information that might 
affect either the ethical and or scientific conduct of the trial, or protocol deviations will be 
immediately reported to the IRB using iRIS. 
The principal investigator will review adverse event and response data on each patient to ensure 
safety and data accuracy. The principal investigator will personally conduct or supervise the 
investigation and provide appropriate delegation of responsibilities to other members of the 
research staff. 
7.5.2
Sponsor Monitoring Plan
As a sponsor for clinical trials, FDA regulations require the CCR to maintain a monitoring 
program. The CCR’s program allows for confirmation of: study data, specifically data that could 
affect the interpretation of primary study endpoints; adherence to the protocol, regulations, and 
SOPs; and human subjects protection. This is done through independent verification of study 
data with source documentation focusing on:
x
Informed consent process
x
Eligibility confirmation
x
Drug administration and accountability
x
Adverse events monitoring
x
Response assessment.

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The monitoring program also extends to multi-site research when the CCR is the coordinating 
center.
This trial will be monitored by personnel employed by a CCR contractor. Monitors are qualified 
by training and experience to monitor the progress of clinical trials. Personnel monitoring this 
study will not be affiliated in any way with the trial conduct. 
7.5.3
Safety Monitoring Committee (SMC)
This protocol will require oversight from the Safety Monitoring Committee (SMC).  Initial 
review will occur as soon as possible after the annual NCI-IRB continuing review date.   
Subsequently, each protocol will be reviewed as close to annually as the quarterly meeting 
schedule permits or more frequently as may be required by the SMC.   For initial and subsequent 
reviews, protocols will not be reviewed if there is no accrual within the review period. Written 
outcome letters will be generated in response to the monitoring activities and submitted to the 
Principal investigator and Clinical Director or Deputy Clinical Director, CCR, NCI.
8
STATISTICAL CONSIDERATIONS
The primary objective of this phase I trial is to determine the safe dose level of E6 TCR T cells 
for the treatment of vulvar HSIL. Secondary objectives include an assessment of the clinical 
responses in subjects receiving this treatment and exploratory analyses of translational research 
endpoints. 
This is a phase I trial with a standard 3+3 design and two dose levels. The maximum number of 
patients required to determine the MTD on this trial is 12. Six additional patients will be 
evaluated at the MTD, for a maximum intended enrollment of 18 patients. To allow for patients 
who enroll but are not evaluable for the primary endpoint and screen failures, up to 200 patients 
total may be enrolled. 
The expected accrual rate is 10-15 patients per year. It should therefore take 1.25-2 years to 
complete accrual of patients.
The clinical responses in 12 patients tested at the MTD may be used in the design of a 
subsequent phase II trial.
9
COLLABORATIVE AGREEMENTS
9.1
COOPERATIVE RESEARCH AND DEVELOPMENT AGREEMENT (CRADA)
A CRADA (# 03022) between NCI and Kite Pharma is in place.  Kite Pharma will be providing 
funding for this study.
10
HUMAN SUBJECTS PROTECTIONS 
10.1 RATIONALE FOR SUBJECT SELECTION
The patients to be entered in this protocol have vulvar HSIL that is unable to be completely
resected to negative margins without causing deformity. As only women develop vulvar HSIL,
men will be excluded from the study.
Subjects from all racial/ethnic groups are eligible for this study if they meet the eligibility
criteria. To date, there is no information that suggests that differences in drug metabolism or
disease response would be expected in one group compared to another. If differences in outcome
that correlate to ethnic identity are noted, accrual may be expanded or a follow-up study may be
written to investigate those differences more fully.

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10.2 STRATEGIES/PROCEDURES FOR RECRUITMENT
Patients for this protocol will be recruited via standard CCR mechanisms as well as various 
advertising venues.  All advertisements, letters and other recruitment efforts will be submitted to 
the IRB for approval prior to their implementation.  
10.3 PARTICIPATION OF CHILDREN
The efficacy of this treatment is unknown, and no safety data in children exist; children are
therefore excluded from participation. In addition, vulvar HSIL is exceedingly rare in children so
a small number of children, if any, would be eligible to participate.
10.4 PARTICIPATION OF SUBJECTS UNABLE TO GIVE CONSENT
Adults unable to give consent are excluded from enrolling in the protocol.  However re-consent 
may be necessary and there is a possibility, though unlikely, that subjects could become 
decisionally impaired. For this reason and because there is a prospect of direct benefit from 
research participation (section 10.6), all subjects will be offered the opportunity to fill in their 
wishes for research and care, and assign a substitute decision maker on the “NIH Advance 
Directive for Health Care and Medical Research Participation” form so that another person can 
make decisions about their medical care in the event that they become incapacitated or 
cognitively impaired during the course of the study. Note: The PI or AI will contact the NIH 
Ability to Consent Assessment Team (ACAT) for evaluation as needed for the following: an 
independent assessment of whether an individual has the capacity to provide consent; assistance 
in identifying and assessing an appropriate surrogate when indicated; and/or an assessment of the 
capacity to appoint a surrogate.  For those subjects that become incapacitated and do not have 
pre-determined substitute decision maker, the procedures described in MEC Policy 87-4 and 
NIH HRPP SOP 14E for appointing a surrogate decision maker for adult subjects who are (a) 
decisionally impaired, and (b) who do not have a legal guardian or durable power of attorney, 
will be followed.
10.5 EVALUATION OF BENEFITS AND RISKS/DISCOMFORTS
The experimental treatment has well-defined risks based on experience with the administration
of E6 TCR T cells and aldesleukin. The benefit of the treatment is unknown. The alternative to
treatment on the protocol is surgical resection, which carries risks of disfiguration, pain,
anesthesia complications, wound infections, functional impairment, and disease recurrence.
The additional risks associated with potential study procedures have been outlined and described
in detail in the consent form.
10.6 RISKS/BENEFITS ANALYSIS (INCLUDING WHO ARE OR MAY BECOME UNABLE TO
CONSENT)
All patients in this protocol have vulvar HSIL, and therefore an increased risk of vulvar cancer. 
In addition, their standard treatment option is surgery with unacceptable cosmetic or functional 
results and a high risk of disease recurrence. The protocol has defined risks based on prior 
studies with E6 TCR T cells and extensive clinical testing of aldesleukin. In this study, 
aldesleukin is administered after cell infusion to promote growth and survival of the infused 
cells. Although aldesleukin can cause serious adverse events, it’s use in this protocol is essential.
The success of this effort cannot be predicted now. Because all patients in this protocol have 
vulvar HSIL and therefore an increased risk of vulvar cancer the potential benefit is thought to 

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outweigh the potential risks. Although, this protocol involves greater than minimal risk, it 
presents the potential for direct benefit to individual subjects.
.
10.7 CONSENT PROCESS AND DOCUMENTATION
The patient, along with family members or friends, will be presented with a detailed description
of the protocol treatment. The specific requirements, objectives, and potential advantages and
disadvantages will be presented. The Informed Consent document is given to the patient who is
requested to review it and to ask questions prior to agreeing to participate in the treatment
portion of this protocol. The patient will be reassured that participation on trial is entirely
voluntary and that she can withdraw or decide against treatment at any time without adverse
consequences. The research nurse, Principal Investigator or his designee is responsible for
obtaining written informed consent from the patient.
If new safety information results in significant changes in the risk/ benefit assessment, the 
consent form will be reviewed and updated as necessary. All subjects (including those already 
being treated) will be informed of the new information, be given a copy of the revised form, and 
asked to give their consent to continue in the study.
For optional biopsies, patients will be required to sign a separate consent for all biopsies 
conducted during the study. If the patient refuses the optional biopsy at that time, the refusal will 
be documented in the medical record and the research record.
10.7.1
Telephone consent and re-consent
Consent for screening and re-consent on this study may be obtained via telephone according to 
the following procedure: the informed consent document will be sent to the subject.  An 
explanation of the screening requirements or change (s) in the study will be provided over the 
telephone after the subject has had the opportunity to read the consent form.  The subject will 
sign and date the informed consent. A witness to the subject’s signature will sign and date the 
consent.  
The original informed consent document will be sent back to the consenting investigator who 
will sign and date the consent form with the date the consent was obtained via telephone.  
A fully executed copy will be returned via mail for the subject’s records.  The informed consent 
process will be documented on a progress note by the consenting investigator.
10.7.2
Enrollment of non-English Speaking Subjects
If there is an unexpected enrollment of a research participant for whom there is no translated 
extant IRB approved consent document, the principal investigator and/or those authorized to 
obtain informed consent will use the Short Form Oral Consent Process as described in MAS 
Policy M77-2, OHSRP SOP 12, 45 CFR 46.117 (b) (2), and 21 CFR 50.27 (b) (2). The 
summary that will be used is the English version of the extant IRB approved consent document.  
Signed copies of both the English version of the consent and the translated short form will be 
given to the subject or their legally authorized representative and the signed original will be filed 
in the medical record.

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Unless the PI is fluent in the prospective subject’s language, an interpreter will be present to 
facilitate the conversation. Preferably someone who is independent of the subject (i.e., not a 
family member) will assist in presenting information and obtaining consent. Whenever possible, 
interpreters will be provided copies of the relevant consent documents well before the consent 
conversation with the subject (24 to 48 hours if possible). 
We request prospective IRB approval of the use of the short form process for non-English 
speaking subjects and will notify the IRB at the time of continuing review of the frequency of the 
use of the Short Form.
11 PHARMACEUTICAL INFORMATION  
11.1 INTERLEUKIN-2 (ALDESLEUKIN, PROLEUKIN, RECOMBINANT HUMAN INTERLEUKIN 2)
11.1.1
How Supplied:
Aldesleukin (interleukin-2) will be provided by the NIH Clinical Pharmacy Department from 
commercial sources.
11.1.2
Formulation/Reconstitution:
Aldesleukin, NSC #373364, is provided as single-use vials containing 22 million IU (~1.3mg) 
IL-2 as a sterile, white to off-white lyophilized cake plus 50mg mannitol and 0.18 mg sodium 
dodecyl sulfate, buffered with approximately 0.17 mg monobasic and 0.89 mg dibasic sodium 
phosphate to a pH of 7.5 (range 7.2 to 7.8).  The vial is reconstituted with 1.2 mL of Sterile 
Water for Injection, USP, and the resultant concentration is 18 million IU/mL or 1.1 mg/mL.  
Diluent should be directed against the side of the vial to avoid excess foaming.  Swirl contents 
gently until completely dissolved.  Do not shake.  Since vials contain no preservative, 
reconstituted solution should be used with 24 hours.
11.1.3
Storage:
Intact vials are stored in the refrigerator (2 to 8C) protected from light.  Each vial bears an 
expiration date.
11.1.4
Dilution/Stability:
Reconstituted aldesleukin should be further diluted with 50 mL of 5% Human Serum Albumin 
(HSA).  The human serum albumin should be in the final container (bottle or bag) prior to the 
addition of aldesleukin.  Dilutions of the reconstituted solution over a 1000-fold range (i.e., 1 
mg/mL to 1 mcg/mL) are acceptable in either glass bottles or polyvinyl chloride bags.  
Aldesleukin is chemically stable for 48 hours at refrigerated and room temperatures, 2 to 30C.
11.1.5
Administration:
The dosage will be calculated based on total body weight.  The final dilution of aldesleukin will 
be infused over 15 minutes.  Aldesleukin will be administered as an inpatient.
11.1.6
Toxicities:
Expected toxicities of aldesleukin are listed in the product label and in Appendix B and 
Appendix C. Grade 3 toxicities common to aldesleukin include diarrhea, nausea, vomiting, 
hypotension, skin changes, anorexia, mucositis, dysphagia, or constitutional symptoms and 
laboratory changes. Additional Grade 3 and 4 toxicities seen with aldesleukin are detailed in 
Appendix B and Appendix C.

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11.2 CELL PREPARATION (E6 TCR TRANSDUCED PBL) (IND # 16078)
The procedure for the expanding the human PBL and the Certificate of Analysis (CoA) are 
similar to those approved by the Food and Drug Administration, and used at the NCI in ongoing 
protocols evaluating cell therapy in the Surgery Branch.  The CoA is included Appendix D. The 
PBL will be transduced with retroviral supernatant containing the E6 TCR.
11.2.1
Retroviral Vector Containing the E6 TCR gene
The retroviral vector supernatant (PG13-MSGV1-E6-TCR) encoding a T cell receptor directed 
against HPV16 E629-36) was prepared and preserved following a cGMP conditions in the Surgery 
Branch Vector Production Facility (SBVPF).  
The retroviral vector E6 TCR consists of 7,310 bps including the 5’LTR from the murine stem 
cell virus (promoter), packaging signal including the splicing donor (SD) and splicing acceptor 
sites, alpha and beta chain genes of the E6 TCR.  The alpha and beta chains are linked by a P2A 
peptide. The vector was codon optimized for expression by human cells with constant region 
exchanged for murine counterparts with an added disulfide bond and hydrophobic substitutions 
in the alpha chain constant region transmembrane domain.    
The physical titer will be determined by transduction of PBL with serial dilutions of the vector.  
TCR expression on the cell surface will be measured using FACS following staining with an 
anti-mouse constant region antibody.  The titer will be measured as transducing units per 
milliliter.  Portions of the supernatant will be stored at -80C at the NIH Clinical Center 
Department of Transfusion Medicine.  These storage facilities are equipped with around-the-
clock temperature monitoring.  Upon request, supernatant will be delivered on dry ice to be used 
in ex vivo transduction of patient PBL.  There will be no re-use of the same unit of supernatant 
for different patients.  Retroviral titer has been shown to be stable after immediate thawing and 
immediate administration (coating the tissue culture wells previously coated with Retronectin).  
Handling of the vector should follow the guidelines of Biosafety Level-2 (BSL-2). The specific 
guidelines for Biosafety Level-2 (BSL-2) can be viewed at http://bmbl.od.nih.gov/sect3bsl2.htm

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12 REFERENCES 
1.
Bornstein, J., et al., The 2015 International Society for the Study of Vulvovaginal Disease 
(ISSVD) Terminology of Vulvar Squamous Intraepithelial Lesions. J Low Genit Tract 
Dis, 2016. 20(1): p. 11-4.
2.
de Sanjose, S., et al., Worldwide human papillomavirus genotype attribution in over 2000 
cases of intraepithelial and invasive lesions of the vulva. Eur J Cancer, 2013. 49(16): p. 
3450-61.
3.
van Seters, M., M. van Beurden, and A.J. de Craen, Is the assumed natural history of 
vulvar intraepithelial neoplasia III based on enough evidence? A systematic review of 
3322 published patients. Gynecol Oncol, 2005. 97(2): p. 645-51.
4.
van de Nieuwenhof, H.P., et al., Vulvar squamous cell carcinoma development after 
diagnosis of VIN increases with age. Eur J Cancer, 2009. 45(5): p. 851-6.
5.
Cendejas, B.R., K.K. Smith-McCune, and M.J. Khan, Does treatment for cervical and 
vulvar dysplasia impact women's sexual health? Am J Obstet Gynecol, 2015. 212(3): p. 
291-7.
6.
Wallbillich, J.J., et al., Vulvar intraepithelial neoplasia (VIN 2/3): comparing clinical 
outcomes and evaluating risk factors for recurrence. Gynecol Oncol, 2012. 127(2): p. 
312-5.
7.
Judson, P.L., et al., Trends in the incidence of invasive and in situ vulvar carcinoma.
Obstet Gynecol, 2006. 107(5): p. 1018-22.
8.
de Witte, C.J., et al., Imiquimod in cervical, vaginal and vulvar intraepithelial neoplasia: 
a review. Gynecol Oncol, 2015. 139(2): p. 377-84.
9.
Kenter, G.G., et al., Vaccination against HPV-16 oncoproteins for vulvar intraepithelial 
neoplasia. N Engl J Med, 2009. 361(19): p. 1838-47.
10.
Hung, C.F., et al., Therapeutic human papillomavirus vaccines: current clinical trials 
and future directions. Expert Opin Biol Ther, 2008. 8(4): p. 421-39.
11.
Stevanovic, S., et al., Complete regression of metastatic cervical cancer after treatment 
with human papillomavirus-targeted tumor-infiltrating T cells. J Clin Oncol, 2015. 
33(14): p. 1543-50.
12.
Fisher, R.I., S.A. Rosenberg, and G. Fyfe, Long-term survival update for high-dose 
recombinant interleukin-2 in patients with renal cell carcinoma. Cancer J Sci Am, 2000. 
6 Suppl 1: p. S55-7.
13.
Rosenberg, S.A., et al., Treatment of 283 consecutive patients with metastatic melanoma 
or renal cell cancer using high-dose bolus interleukin 2. JAMA, 1994. 271(12): p. 907-
13.
14.
Klebanoff, C.A., et al., Determinants of successful CD8+ T-cell adoptive immunotherapy 
for large established tumors in mice. Clin Cancer Res, 2011. 17(16): p. 5343-52.
15.
Overwijk, W.W., et al., Tumor regression and autoimmunity after reversal of a 
functionally tolerant state of self-reactive CD8+ T cells. J Exp Med, 2003. 198(4): p. 
569-80.

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16.
Schwartzentruber, D.J., Guidelines for the safe administration of high-dose interleukin-2.
J Immunother, 2001. 24(4): p. 287-93.
17.
Marroquin, C.E., et al., Decreased tolerance to interleukin-2 with repeated courses of 
therapy in patients with metastatic melanoma or renal cell cancer. J Immunother, 2000. 
23(3): p. 387-92.
18.
Rosenberg, S.A., et al., Treatment of patients with metastatic melanoma with autologous 
tumor-infiltrating lymphocytes and interleukin 2. J Natl Cancer Inst, 1994. 86(15): p. 
1159-66.
19.
Rosenberg, S.A., et al., Prospective randomized trial of high-dose interleukin-2 alone or 
in conjunction with lymphokine-activated killer cells for the treatment of patients with 
advanced cancer. J Natl Cancer Inst, 1993. 85(8): p. 622-32.
20.
Scholler, J., et al., Decade-long safety and function of retroviral-modified chimeric 
antigen receptor T cells. Sci Transl Med, 2012. 4(132): p. 132ra53.

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13 APPENDICES
13.1 APPENDIX A -PERFORMANCE STATUS CRITERIA
ECOG Performance Status Scale*
Grade
Descriptions
0
Normal activity.  Fully active, able to carry on all pre-disease 
performance without restriction.
1
Symptoms, but ambulatory.  Restricted in physically strenuous activity, 
but ambulatory and able to carry out work of a light or sedentary nature 
(e.g., light housework, office work).
2
In bed <50% of the time.  Ambulatory and capable of all self-care, but 
unable to carry out any work activities.  Up and about more than 50% 
of waking hours.
3
In bed >50% of the time.  Capable of only limited self-care, confined to 
bed or chair more than 50% of waking hours.
4
100% bedridden.  Completely disabled.  Cannot carry on any self-care.  
Totally confined to bed or chair.
5
Dead.
* As published in Am. J. Clin. Oncol.: Oken, M.M., Creech, R.H., Tormey, D.C., Horton, J., 
Davis, T.E., McFadden, E.T., Carbone, P.P.: Toxicity And Response Criteria Of The Eastern 
Cooperative Oncology Group. Am J Clin Oncol 5:649-655, 1982.

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13.2 APPENDIX B: ADVERSE EVENTS OCCURRING IN  OF PATIENTS TREATED WITH 
ALDESLEUKIN (N=525)1
Body System
% Patients 
Body System
% Patients
Body as a Whole
Metabolic and Nutritional Disorders
Chills 
52
Bilirubinemia 
40
Fever 
29
Creatinine increase 
33
Malaise 
27
Peripheral edema 
28
Asthenia 
23
SGOT increase 
23
Infection 
13
Weight gain 
16
Pain 
12
Edema 
15
Abdominal pain 
11
Acidosis 
12
Abdomen enlarged 
10
Hypomagnesemia 
12
Cardiovascular 
Hypocalcemia 
11
Hypotension 
71
Alkaline phosphatase incr
10
Tachycardia 
23
Nervous
Vasodilation 
13
Confusion 
34
Supraventricular 
12
Somnolence 
22
tachycardia
Cardiovascular disordera
11
Anxiety 
12
Arrhythmia 
10
Dizziness 
11
Digestive 
Respiratory
Diarrhea 
67
Dyspnea 
43
Vomiting 
50
Lung disorderb
24
Nausea 
35
Respiratory disorderc
11
Stomatitis 
22
Cough increase 
11
Anorexia 
20
Rhinitis 
10
Nausea and vomiting 
19
Skin and Appendages
Hemic and Lymphatic
Rash 
42
Thrombocytopenia 
37
Pruritus 
24
Anemia 
29
Exfoliative dermatitis 
18
Leukopenia
16
Urogenital
Oliguria 
63
a Cardiovascular disorder: fluctuations in blood pressure, asymptomatic ECG changes, CHF.
b Lung disorder: physical findings associated with pulmonary congestion, rales, rhonchi.
c Respiratory disorder: ARDS, CXR infiltrates, unspecified pulmonary changes.
1Source: Proleukin® Prescribing Information – June 2007

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54
13.3
APPENDIX C: EXPECTED ALDESLEUKIN TOXICITIES AND THEIR MANAGEMENT
Toxicity
Grade
Supportive Medications
Considered 
DLT*
Chills
3
IV Meperidine 25-50mg IV 
q1hr, prn
No 
Fever
3
Acetaminophen 650mg po 
q4hr; Indomethicin 50-75mg 
po q8h
No
Pruritus
3
Hydroxyzine HCl 10-20mg po 
q6h, prn; Diphenhydramine 
HCl 25-50mg po q4h prn
No 
Nausea/Vomiting/A
norexia
3
Ondansetron 10mg IV q8hr 
prn, Granisetron 0.01 mg/kg 
IV qday prn, Droperidol 1mg 
IV a4-6h prn; Prochlorperazine 
25mg PR prn or 10mg IV q6hr 
prn
No
Diarrhea
3
Loperamide 2mg po q3h prn; 
Diphenoxylate HCl 2.5mg and 
Atropine sulfate 25mcg po q3h 
prn; Codeine sulfate 30-60mg 
po q4h prn
If uncontrolled 
after 48h despite 
all supportive 
measures
Malaise
3 or 4
Bedrest
If other toxicities 
occur 
simultaneously
Hyperbilirubinemia
3 or 4
Observation
Yes
Anemia
3 or 4 
Transfusion with PRBCs
Yes
Thrombocytopenia
3 or 4 
Transfusion with platelets
Yes
Edema/Weight gain
3
Diuretics prn
No 
Hypotension
3
Fluid resuscitation, 
Vasopressor support
No, unless 
requires more 
than 1.5L/24 
hours of bolus 
IV fluids or 
vasopressor 
support
Dyspnea
3 or 4 
Oxygen or ventilator support
If requires more 
than nasal 
cannula oxygen 
support or 
requires oxygen 
for greater than 
48 hours after 
last dose

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55
Toxicity
Grade
Supportive Medications
Considered 
DLT*
Oliguria
3 or 4 
Fluid boluses
If requires more 
than 1.5L/24 
hours of bolus 
IV fluids
Increased Creatinine
3 or 4
Observation
If grade 3 lasts 
more than 48 
hours or for any 
grade 4.
Renal Failure
3 or 4 
Dialysis/CVVH
Yes
Pleural Effusion
3
Thoracentesis
Yes
Bowel Perforation
3
Surgical intervention
Yes
Confusion
3
Observation
Yes
Somnolence
3 or 4 
Intubation for airway 
protection
Yes
Arrhythmia
3
Correction of fluid and 
electrolyte imbalances; 
chemical conversion or 
electrical conversion therapy
Yes
Elevated Troponin 
Levels
3 or 4 
Observation
Yes
Myocardial 
Infarction
4
Supportive care
Yes
Elevated 
Transaminases
3 or 4
Observation
Yes
Hyperbilirubinemia
3 or 4 
Observation
Yes
Electrolyte 
Imbalances
3 or 4
Electrolyte replacement
If uncontrolled 
after 48 hours 
despite all 
supportive 
measures
Neutropenia
4
Observation
Yes
*Unless the toxicity is not reversed within 24 hours of last aldesleukin, in which case 
all AEs are considered DLTs.

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13.4 APPENDIX D: CERTIFICATE OF ANALYSIS HPV-16 E6 TCR
Date of preparation of final product:
Patient:
Tests performed on final product:
Test
Method
Limits
Results 
Initials/Date
Cell viability1
Trypan blue
exclusion
>70%
Total viable cell
number1
Visual microscopic
count
>1x109
Tumor reactivity3
Gamma-IFN
release vs peptide
pulsed T2 cells
>200 pg/mL
and > 2 times
background
TCR expression2
FACS analysis of
the transduced cells
PBL, >10%
Microbiological
studies
Gram stain1,3
No micro-
organisms
seen
Aerobic culture3,4
No growth
Fungal culture3,4
No growth
Anaerobic culture3,4 No growth
Mycoplasma test5
No growth
Endotoxin
Limulus assay1
<5 E.U./kg
RCR
S+L- Assay4
RCR-PCR6
Negative
1Performed on sample of the final product immediately prior to infusion.  Results are available at 
the time of infusion.
2Performed 2 to 10 days post-transduction.  Results are available at the time of infusion.

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3Performed 2 to 4 days prior to infusion.  Results are available at the time of infusion but may 
not be definitive.
4Sample collected from the final product prior to infusion.  Results will not be available before 
cells are infused into the patient.
5Performed 2 to 10 days prior to infusion.  Results are available at the time of infusion.
6Performed on sample approximately 1 to 4 days prior to infusion.  Results are available at the 
time of infusion.
Prepared by:  _________________________  Date:______________________
QC sign-off:  _________________________  Date: ______________________
Qualified Clinical or Laboratory Supervisor

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13.5 APPENDIX E: HPV-16 DDPCR PERIPHERAL BLOOD PROCESSING INSTRUCTIONS TO 
PATIENT AND PHYSICIAN
PATIENT INSTRUCTIONS:
Please take this instruction sheet, along with the sample tube that was sent to you, to your 
physician or the laboratory that will be drawing these labs.
PHYSICIAN/LABORATORY INSTRUCTIONS:
1. Draw peripheral blood into one 10 mL Cell-Free DNA BCT tube (provided). 
2. After blood draw, immediately mix the tube by gentle inversion 8 to 10 times.  
3. Do not vortex tubes, as cellular lysis could occur.  
4. Keep the tubes at 6o C to 37oC.  Do NOT refrigerate or freeze.  
5. Ship the tube via FedEx at 6o C to 37oC to ensure the arrival of tubes within 5 days of 
date of blood draw.  
6. Once shipped, notify Erin Ferraro at erin.ferraro@nih.gov
Address for Shipping:  
Erin Ferraro, RN
National Institutes of Health
10 Center Drive
Building 10-CRC Room 3-3330
Bethesda, MD 20814