Clinical trial NCT02111850
Conditions: Cervical Cancer Renal Cancer Urothelial Cancer Melanoma Breast Cancer
Keywords: Gene Therapy Tumor Regression Immunotherapy Adoptive Cell Therapy
Sponsor: National Cancer Institute (NCI)
Start date: 2014-02-07; Completion date: 2021-03-24
Enrollment: 21 participants; Allocation: NON_RANDOMIZED; Intervention model: SEQUENTIAL

Brief summary:
Background:

The National Cancer Institute (NCI) Surgery Branch has developed an experimental therapy for treating patients with metastatic cancer that involves taking white blood cells from the patient, growing them in the laboratory in large numbers, genetically modifying these specific cells with a type of virus (retrovirus) to attack only the tumor cells, and then giving the cells back to the patient. This type of therapy is called gene transfer. In this protocol, we are modifying the patient s white blood cells with a retrovirus that has the gene for anti-Melanoma antigen family A, 3 (MAGE-A3)-DP0401/0402 incorporated in the retrovirus.

Objective:

The purpose of this study is to determine a safe number of these cells to infuse and to see if these particular tumor-fighting cells (anti-MAGE-A3-DP0401/0402 cells) cause tumors to shrink and to be certain the treatment is safe.

Eligibility:

\- Adult's age 18-70 with metastatic cancer expressing the MAGE-A3 molecule.

Design:

* Work up stage: Patients will be seen as an outpatient at the National Institutes of Health (NIH) clinical Center and undergo a history and physical examination, scans, x-rays, lab tests, and other tests as needed
* Leukapheresis: If the patients meet all of the requirements for the study, they will undergo leukapheresis to obtain white blood cells to make the anti-MAGE-A3-DP0401/0402 cells. {Leukapheresis is a common procedure, which removes only the white blood cells from the patient.}
* Treatment: Once their cells have grown, the patients will be admitted to the hospital for the conditioning chemotherapy, the anti-MAGE-A3-DP0401/0402 cells and aldesleukin. They will stay in the hospital for approximately 4 weeks for the treatment.
* Follow up: Patients will return to the clinic for a physical exam, review of side effects, lab tests, and scans about every 1-3 months for the first year, and then every 6 months to 1 year as long as their tumors are shrinking.

Detailed description:
Background:

* We have constructed a single retroviral vector that contains both and $ \<= chains of a T cell receptor (TCR) that recognizes the DP0401/0402 restricted Melanoma antigen family A, 3 (MAGE-A3) tumor antigen, which can be used to mediate genetic transfer of this TCR with high efficiency.
* In co-cultures with HLA-DP0401/0402 and MAGE-A3 double positive tumors, the anti- MAGE-A3- DP0401/0402 restricted (anti-MAGE-A3-DP4) TCR transduced T cells secreted significant amounts of Interferon gamma (IFN-y) with high specificity.

Objectives:

Primary objectives:

* Determine a safe dose of the administration of autologous cluster of differentiation 4 (CD4) cells transduced with an anti-MAGE-A3-DP0401/0402 restricted (MAGE-A3-DP4) TCR and aldesleukin to patients following a nonmyeloablative but lymphoid depleting preparative regimen.
* Determine if this approach will result in objective tumor regression in patients with metastatic cancer expressing MAGE-A3-DP4.
* Determine the toxicity profile of this treatment regimen.

Eligibility:

Patients who are human leukocyte antigens (HLA)-DP0401/0402 positive and 18 years of age or older must have

* Metastatic cancer whose tumors express the MAGE-A3-DP4 antigen.
* Previously received and have been a non-responder to or recurred following at least one first line treatment for metastatic disease.

Patients may not have:

\- Contraindications for high dose aldesleukin administration.

Design:

* PBMC obtained by leukapheresis will be enriched for CD4 cells and transduced with the retroviral vector supernatant encoding the anti-MAGE-A3-DP4 TCR.
* The study will begin in a standard phase 1 dose escalation. After the maximum tolerated dose (MTD) cell dose has been determined, patients will be enrolled into the phase 2 portion of the trial at the MTD established during the phase 1 portion of the study. In the phase 2 portion, patients will be entered into two cohorts: cohort 1 will include patients with metastatic melanoma; cohort 2 will include patients with renal cancer and other types of metastatic cancer.
* Patients will receive a nonmyeloablative but lymphocyte depleting preparative regimen consisting of cyclophosphamide and fludarabine followed by intravenous infusion of ex vivo tumor reactive, TCR gene-transduced peripheral blood mononuclear cells (PBMC) plus intravenous (IV) aldesleukin.
* Patients will undergo complete evaluation of tumor response every 1-6 months until off study criteria are met.
* For each of the 2 strata evaluated in the phase 2 portion, the study will be conducted using a phase 2 optimal design where initially 21 evaluable patients will be enrolled. For each of these two arms of the trial, if 0 or 1 of the 21 patients experiences a clinical response, then no further patients will be enrolled but if 2 or more of the first 21 evaluable patients enrolled have a clinical response, then accrual will continue until a total of 41 evaluable patients have been enrolled in that stratum.
* For both strata, the objective will be to determine if the treatment regimen is able to be associated with a clinical response rate that can rule out 5% (p0=0.05) in favor of a modest 20% partial response (PR) + complete response (CR) rate (p1=0.20).

Arms: arm 1: Non-myeloablative lymphodepleting preparative regimen of cyclophosphamide and fludarabine + Anti-Melanoma antigen family A, 3 (MAGE-A3)-DP4 T cell receptor (TCR) peripheral blood lymphocytes (PBL) + high-dose aldesleukin arm 2: Non-myeloablative lymphodepleting preparative regimen of cyclophosphamide and fludarabine + Anti-Melanoma antigen family A, 3 (MAGE-A3)-DP4 T cell receptor (TCR) peripheral blood lymphocytes (PBL) + high-dose aldesleukin
Interventions: intervention 1: Anti-MAGE-A3-DP4 T Cell Receptor (TCR) Peripheral Blood Lymphocytes (PBL) intervention 2: Cyclophosphamide intervention 3: Fludarabine intervention 4: Aldesleukin
Intervention descriptions: intervention 1: Day 0: cells will be infused intravenously (i.v.) on the Patient Care Unit over 20 to 30 minutes intervention 2: Days -7 and -6: Cyclophosphamide 60 mg/kg/day X 2 days intravenous (IV) in 250 ml dextrose 5 % in water (D5W) with Mesna 15 mg/kg /day X 2 days over 1 hour. intervention 3: Days -7 to -3: Fludarabine 25 mg/m\^2 /day intravenous piggy-back (IVPB) daily over 30 minutes for 5 days. intervention 4: Aldesleukin 720,000 IU/kg intravenous (IV) (based on total body weight) over 15 minutes every eight hours (+/- 1 hour) beginning within 24 hours of cell infusion and continuing for up to 5 days (maximum 15 doses).

Protocol documents:
Prot_SAP_000.pdf: 
Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
1
Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
CC Protocol Number: 14-C-0052 K 
IBC Number: RD-13-X-03 
OSP Number: 1310-1253 
NCT Number: NCT02111850 
Version Date: June 22, 2018  
PROTOCOL TITLE 
A Phase I/II Study of the Treatment of Metastatic Cancer that Expresses MAGE-A3 Using 
Lymphodepleting Conditioning Followed by Infusion of HLA-DP0401/0402 Restricted Anti-
MAGE-A3 TCR-Gene Engineered Lymphocytes and Aldesleukin 
 
NIH Principal Investigator:  
Steven A. Rosenberg, M.D., Ph.D.  
 
 
 
 
 
Chief of Surgery, Surgery Branch, NCI 
 
Building 10, CRC, Room 3-3940 
 
9000 Rockville Pike, Bethesda, MD 20892 
 
 
 
 
 
Phone: 240-760-6218; Email: Steven.Rosenberg@nih.gov  
 
  
Investigational Agent: 
 
Drug Name: 
Anti-MAGE-A3-DP4 TCR PBL 
IND Number: 
15770 
Sponsor: 
Center for Cancer Research 
Manufacturer: 
Surgery Branch Cell Production Facility 
 
 
Commercial Agents: Cyclophosphamide, Fludarabine, and Aldesleukin 
 
 
 


A b brevi ate d Title: A nti -M A G E -A 3 -D P 4 T C R P B L  
Versi o n D ate: J u ne 2 2, 2 0 1 8  
 
2  
P R É CI S  
B ac k gr o u n d: 
•  We ha ve c o nstr ucte d a si n gle retr o viral vect or t hat c o ntai ns b ot h α a n d β c hai ns of a T 
cell rece pt or ( T C R) t hat rec o g nizes t he D P 0 4 0 1 / 0 4 0 2 restricte d M A G E-A 3 t u m or 
a nti ge n, w hi c h ca n be use d t o me diate ge netic tra nsfer of t his T C R wit h hi g h efficie nc y.  
•  I n c o-c ult ures wit h H L A - D P 0 4 0 1/ 0 4 0 2 a n d M A G E- A 3 d o u ble p ositi ve t u m ors, t he a nti-
M A G E -A 3 - D P 0 4 0 1/ 0 4 0 2 restricte d (a nti-M A G E -A 3 - D P 4) T C R tra ns d uce d T cells 
secrete d si g nifica nt a mo u nts of I F N -γ( wit h hi g h s pecificit y.  
O bjecti ves : 
•  Pri mar y o bjecti ves : 
o  D eter mi ne a s afe d os e of t he a d mi nistrati o n of a ut ol o g o us C D 4 cells tra ns d uce d wit h 
a n a nti-M A G E -A 3 - D P 0 4 0 1/ 0 4 0 2 restricte d ( M A G E-A 3 -D P 4) T C R  a n d al desle u ki n 
t o patie nts f oll o wi n g a n o n m yel oa blati ve b ut l y m p h oi d de pleti n g pre parati ve re gi me n. 
o  D eter mi ne if t his a p pr oac h will res ult i n o bjecti ve  t u m or re gressi o n i n patie nts wit h 
metastatic ca ncer e x pressi n g M A G E- A 3 -D P 4 . 
o  D eter mi ne t he t o xicit y pr ofile of t his treat me nt re gi me n.  
Eli gi bili t y: 
Patie nts w h o are H L A - D P 0 4 0 1/ 0 4 0 2 p ositi ve a n d 1 8 years of a ge or ol der m ust ha ve: 
•  Metastatic ca ncer w h os e t u m ors e x press t he M A G E -A 3 -D P 4 a nti ge n;  
•  Pre vi o usl y recei ve d a n d ha ve bee n a n o n-res p o n der t o or rec urre d f oll o wi n g at least o ne 
first li ne treat me nt f or metastatic dis ease;  
Patie nts ma y n ot ha ve:  
•  C o ntrai n dicati o ns f or hi g h d os e al desle u ki n a d mi nistrati o n. 
Desi g n : 
•  P B M C o btai ne d b y le u ka p heresis will be e nric he d f or C D 4 cells a n d tra ns d uce d wit h t he 
retr o viral vect or s u per nata nt e nc o di n g t he a nti-M A G E -A 3 -D P 4 T C R.  
•  T he st u d y will be gi n i n a sta n dar d p has e 1 d os e escalati o n . After t he M T D cell d os e has 
bee n deter mi ne d, patie nts will be e nr olle d i nt o t he p has e 2 p orti o n of t he trial at t he M T D 
esta blis he d d uri n g t he p has e 1 p orti o n of t he st u d y.  I n t he p hase 2 p orti o n, patie nts will be 
e ntere d i nt o t w o c o h orts: c o h ort 1 will i ncl u de patie nts wit h metastatic mela n o ma; c o h ort 
2 will i ncl u de patie nts wit h re nal ca ncer a n d ot her t y pes o f metastatic ca ncer.  
•  Patie nts will recei ve a n o n m yel oa blati ve b ut l y m p h oc yte de pleti n g pre parati ve re gi me n 
c o nsisti n g of c ycl o p h os p ha mi de a n d fl u dara bi ne f oll o we d b y i ntra ve n o us i nf usi o n of e x 
vi v o t u m or reacti ve, T C R ge ne -tra ns d uce d P B M C pl us I V al desle uki n.   
•  Patie nts will u n der g o c o m plete e val uati o n of t u m or res p o nse e ver y 1 - 6 m o nt hs u ntil off 
st u d y criteria are met. 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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• For each of the 2 strata evaluated in the phase 2 portion, the study will be conducted 
using a phase 2 optimal design where initially 21 evaluable patients will be enrolled. For 
each of these two arms of the trial, if 0 or 1 of the 21 patients experiences a clinical 
response, then no further patients will be enrolled but if 2 or more of the first 21 
evaluable patients enrolled have a clinical response, then accrual will continue until a 
total of 41 evaluable patients have been enrolled in that stratum.  
• For both strata, the objective will be to determine if the treatment regimen is able to be 
associated with a clinical response rate that can rule out 5% (p0=0.05) in favor of a 
modest 20% PR + CR rate (p1=0.20).  
 
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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TABLE OF CONTENTS  
PRÉCIS .......................................................................................................................................... 2 
1 
INTRODUCTION ................................................................................................................. 8 
1.1 
Study Objectives ................................................................................................................. 8 
1.1.1 
Primary Objectives........................................................................................................ 8 
1.1.2 
Secondary Objective ..................................................................................................... 8 
1.2 
Background and Rationale .................................................................................................. 8 
1.2.1 
Surgery Branch Trials of Cell Transfer Therapy Using Transduction of Anti-Tumor 
Antigen TCR Genes into PBL ...................................................................................... 8 
1.2.2 
MAGE-A3 as a target for cell transfer clinical studies ................................................. 9 
1.2.3 
Rationale for Targeting a Class II Restricted Epitope of MAGE-A3 recognized by 
CD4 T Cells ................................................................................................................ 11 
1.2.4 
Safety Considerations ................................................................................................. 14 
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 
Within 3 Months Prior to Enrollment ......................................................................... 16 
2.2.2 
Within 8 Weeks Prior to Enrollment: ......................................................................... 16 
2.2.3 
Within 4 Weeks Prior to Enrollment: ......................................................................... 17 
2.2.4 
Within 14 Days Prior to Enrollment: .......................................................................... 17 
2.2.5 
Within 7 Days Prior to Enrollment: ............................................................................ 17 
2.3 
Registration and Treatment Assignment Procedures ........................................................ 17 
2.3.1 
Prior to registration for this protocol .......................................................................... 17 
2.3.2 
Registration procedure ................................................................................................ 17 
2.3.3 
Treatment Assignment Procedures ............................................................................. 17 
3 
STUDY IMPLEMENTATION .......................................................................................... 18 
3.1 
Study Design ..................................................................................................................... 18 
3.1.1 
Performed on 03-C-0277 ............................................................................................ 18 
3.1.2 
Treatment Phase .......................................................................................................... 18 
3.1.3 
Dose Limiting Toxicity: .............................................................................................. 19 
3.1.4 
Phase I - Dose Escalation ............................................................................................ 19 
3.1.5 
Phase 2 Portion ........................................................................................................... 20 
3.1.6 
Protocol Stopping Rules ............................................................................................. 20 
3.2 
Drug Administration ......................................................................................................... 20 
3.2.1 
Preparative Regimen with Cyclophosphamide and Fludarabine ................................ 20 
3.2.2 
Cell Infusion and Aldesleukin Administration: .......................................................... 21 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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3.2.3 
Treatment Schedule .................................................................................................... 22 
3.3 
On-Study Evaluations ....................................................................................................... 23 
3.3.1 
Within 14 days prior to starting the preparative regimen ........................................... 23 
3.3.2 
During the preparative regimen: DAILY .................................................................... 23 
3.3.3 
After Cell Infusion: ..................................................................................................... 23 
3.3.4 
During Hospitalization: ............................................................................................... 23 
3.4 
Retreatment ....................................................................................................................... 24 
3.5 
Post Treatment Evaluation ................................................................................................ 24 
3.5.1 
Time period of evaluations ......................................................................................... 24 
3.5.2 
24 
3.6 
Criteria for Removal from Protocol Therapy and Off Study Criteria ............................... 25 
3.6.1 
Criteria for removal from protocol therapy ................................................................ 25 
3.6.2 
Off Study Criteria ....................................................................................................... 25 
4 
CONCOMITANT MEDICATIONS/MEASURES .......................................................... 26 
4.1 
Infection Prophylaxis: ....................................................................................................... 26 
4.1.1 
Pneumocystis Jirovecii Pneumonia............................................................................. 26 
4.1.2 
Herpes or Varicella Zoster Virus Prophylaxis ............................................................ 26 
4.1.3 
Fungal Prophylaxis ..................................................................................................... 26 
4.1.4 
Empiric Antibiotics ..................................................................................................... 26 
4.2 
Blood Product Support ...................................................................................................... 27 
4.3 
Other Concomitant medications to control side effects .................................................... 27 
5 
BIOSPECIMEN COLLECTION ...................................................................................... 27 
5.1 
Correlative Studies for Research: ..................................................................................... 27 
5.2 
Samples Sent To Dr. Figg’s Lab ....................................................................................... 27 
5.3 
Samples Sent To Surgery Branch cell Processing Laboratory ......................................... 27 
5.3.1 
Prior to chemotherapy administration:........................................................................ 28 
5.3.2 
Prior to cell infusion (one 8ml SST): .......................................................................... 28 
5.3.3 
Post cell infusion evaluations: .................................................................................... 28 
5.4 
Immunological Testing: .................................................................................................... 28 
5.5 
Monitoring Gene Therapy Trials: Persistence and RCR: ................................................. 28 
5.6 
Sample Storage, Tracking and Disposition For SB cell Processing laboratory ................ 29 
5.7 
Sample Storage, Tracking and Disposition For Dr. Figg’s Lab ....................................... 29 
5.7.1 
Sample Data Collection .............................................................................................. 29 
5.7.2 
Sample Storage and Destruction ................................................................................. 30 
6 
DATA COLLECTION AND EVALUATION .................................................................. 30 
6.1 
Data Collection ................................................................................................................. 30 
6.1.1 
Exclusions to Routine Adverse Event Recording: ...................................................... 31 
6.2 
Data Sharing Plan ............................................................................................................. 31 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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6.2.1 
Human Data Sharing Plan ........................................................................................... 31 
6.2.2 
Genomic Data Sharing Plan ........................................................................................ 32 
6.3 
Response Criteria .............................................................................................................. 32 
6.3.1 
Definitions................................................................................................................... 32 
6.3.2 
Disease Parameters ..................................................................................................... 32 
6.3.3 
Methods for Evaluation of Measurable Disease ......................................................... 33 
6.3.4 
Response Criteria ........................................................................................................ 34 
6.3.5 
Confirmatory Measurement/Duration of Response .................................................... 35 
6.4 
Toxicity Criteria ................................................................................................................ 35 
7 
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN .................................................................................................................................... 36 
7.1 
Definitions......................................................................................................................... 36 
7.1.1 
Adverse Event ............................................................................................................. 36 
7.1.2 
Suspected adverse reaction ......................................................................................... 36 
7.1.3 
Unexpected adverse reaction ...................................................................................... 36 
7.1.4 
Serious......................................................................................................................... 36 
7.1.5 
Serious Adverse Event ................................................................................................ 36 
7.1.6 
Disability ..................................................................................................................... 36 
7.1.7 
Life-threatening adverse drug experience ................................................................... 37 
7.1.8 
Protocol Deviation (NIH Definition) .......................................................................... 37 
7.1.9 
Non-compliance (NIH Definition) .............................................................................. 37 
7.1.10 Unanticipated Problem................................................................................................ 37 
7.2 
NCI-IRB and Clinical Director (CD) Reporting ............................................................... 37 
7.2.1 
NCI-IRB and NCI CD Expedited Reporting of Unanticipated Problems and Deaths 37 
7.2.2 
NCI-IRB Requirements for PI Reporting at Continuing Review ............................... 37 
7.2.3 
NCI-IRB Reporting of IND Safety Reports................................................................ 38 
7.3 
IND Sponsor Reporting Criteria ....................................................................................... 38 
7.3.1 
Waivier of expedited reporting to CCR ...................................................................... 38 
7.3.2 
Reporting Pregnancy ................................................................................................... 39 
7.4 
Institutional Biosafety Committee (IBC) Reporting Criteria ............................................ 39 
7.4.1 
Serious Adverse Event Reports to IBC ....................................................................... 39 
7.4.2 
Annual Reports to IBC................................................................................................ 40 
7.5 
Data and Safety Monitoring Plan ...................................................................................... 40 
7.5.1 
Principal Investigator/Research Team ........................................................................ 40 
7.5.2 
Sponsor Monitoring Plan ............................................................................................ 41 
7.5.3 
Safety Monitoring Committee (SMC) ........................................................................ 41 
8 
STATISTICAL CONSIDERATIONS ............................................................................... 41 
9 
COLLABORATIVE AGREEMENTS .............................................................................. 43 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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10 HUMAN SUBJECTS PROTECTIONS ............................................................................ 43 
10.1 
Rationale For Subject Selection ........................................................................................ 43 
10.2 
Participation of Children ................................................................................................... 43 
10.3 
Participation of Subjects Unable to Give Consent ............................................................ 43 
10.4 
Evaluation of Benefits and Risks/Discomforts ................................................................. 44 
10.5 
Risk/Benefit Analysis ....................................................................................................... 44 
10.6 
Consent Process and Documentation ................................................................................ 44 
10.6.1 Informed consent of non-English speaking subjects: ................................................. 44 
11 PHARMACEUTICAL INFORMATION ......................................................................... 45 
11.1 
Investigational Investigation ............................................................................................. 45 
11.1.1 Cell Preparation (ANTI-MAGE-A3-DP4 TRANSDUCED PBL) ............................. 45 
11.1.2 Interleukin-2 (Aldesleukin, Proleukin, Recombinant Human Interleukin 2) ............. 45 
11.1.3 Fludarabine ................................................................................................................. 46 
11.1.4 Cyclophosphamide ...................................................................................................... 47 
11.2 
Supportive Medications .................................................................................................... 48 
11.2.1 Mesna .......................................................................................................................... 48 
11.2.2 Filgrastim .................................................................................................................... 48 
11.2.3 Trimethoprim and Sulfamethoxazole Double Strength (TMP / SMX DS): ............... 48 
11.2.4 Herpes and Varicella Zoster Virus Prophylaxis.......................................................... 49 
11.2.5 Fluconazole ................................................................................................................. 50 
11.2.6 Ondansetron hydrochloride ......................................................................................... 50 
11.2.7 Furosemide .................................................................................................................. 50 
12 REFERENCES .................................................................................................................... 51 
13 FIGURES, TABLES & APPENDICES ............................................................................. 56 
 
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
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1 
INTRODUCTION 
1.1 
STUDY OBJECTIVES 
1.1.1 Primary Objectives 
• Determine a safe dose of the administration of autologous CD4 cells transduced with an 
anti-MAGE-A3-DP0401/0402 restricted (MAGE-A3-DP4) TCR and aldesleukin to 
patients following a nonmyeloablative but lymphoid depleting preparative regimen. 
• Determine if this approach will result in objective tumor regression in patients with 
metastatic cancer expressing MAGE-A3-DP4. 
• Determine the toxicity profile of this treatment regimen. 
1.1.2 Secondary Objective 
• Determine the in vivo survival of TCR gene-engineered cells. 
1.2 
BACKGROUND AND RATIONALE 
Studies in experimental animals have demonstrated that the cellular rather than the humoral arm 
of the immune response plays the major role in the elimination of murine tumors. Much of this 
evidence was derived from studies in which the adoptive transfer of T lymphocytes from 
immune animals could transfer resistance to tumor challenge or in some experiments, the 
elimination of established cancer. Thus, most strategies for the immunotherapy of patients with 
cancer have been directed at stimulating strong T cell immune reactions against tumor-associated 
antigens. 
In contrast to antibodies that recognize epitopes on intact proteins, T cells recognize short 
peptide fragments (8-18 amino acids) that are presented on surface class I or II major 
histocompatibility (MHC) molecules and it has been shown that tumor antigens are presented 
and recognized by T cells in this fashion. The molecule that recognizes these peptide fragments 
is the T-cell receptor (TCR). The TCR is analogous to the antibody immunoglobulin molecule in 
that, two separate proteins (the TCR alpha and beta chains) are brought together to form the 
functional TCR molecule. The goal of this protocol is to transfer MAGE-A3 reactive TCR genes 
into normal peripheral blood lymphocytes (PBL) derived from cancer patients and to return these 
engineered cells to patients aimed at mediating regression of their tumors. This trial is similar to 
previous Surgery Branch TCR gene transfer adoptive immunotherapy protocols that have used 
HLA-Class I restricted TCRs except that in this trial we will use an HLA-Class II 
(DP0401/0402) restricted TCR reactive with a MAGE-A3 epitope expressed on the tumors of 
patients with melanoma as well as patients with common epithelial malignancies. 
1.2.1 Surgery Branch Trials of Cell Transfer Therapy Using Transduction of Anti-Tumor 
Antigen TCR Genes into PBL 
We have studied approaches to transduce genes encoding antigen specific TCRs genes into PBL 
as a method to generate large numbers of reactive anti-cancer T cells. 
In earlier studies we treated 24 patients with metastatic melanoma using autologous PBL 
transduced with high-avidity MART-1 F5 TCR following a non-myeolablative chemotherapy (1, 
2). Six patients (25%) achieved an objective partial response though 15 patients developed a 
transient mild anterior uveitis easily reversed by steroid eye drops and ten patients developed 
decreased hearing reversed by middle ear steroid injections. Transient rashes have also been 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
9
seen. We have also conducted a clinical trial with a TCR that recognizes the gp100:154-162 
melanoma peptide(2). This TCR was raised in an HLA-A2 transgenic mouse immunized with this 
peptide. We treated 21 patients with metastatic melanoma using autologous PBL transduced with 
this gp100 TCR following a non-myeloablative chemotherapy. Four patients (19%) achieved an 
objective partial response. Seven patients developed a transient mild anterior uveitis reversed by 
steroid eye drops and ten patients developed decreased hearing reversed by middle ear steroid 
injections. There were no treatment related deaths in the trial. We are no longer using these 
TCRs in clinical studies. 
At the time of this protocol, we have performed several additional studies utilizing peripheral 
blood lymphocytes transduced with either TCR or chimeric antigen receptors (CAR) genes 
targeting putative tumor antigens following a non-myeloablative chemotherapy regimen. In 
protocol 08-C-0121, 10 0f 19 patients (53%) with metastatic melanoma treated with autologous 
PBL genetically engineered to express an anti-NY-ESO-1 TCR experienced objective responses 
including four patients with complete responses, three of which are ongoing from 24 to 50 plus 
months(3). Similarly, 15 patients with metastatic synovial cell sarcoma also received autologous 
genetically engineered cells expressing the anti-NY-ESO-1 TCR and 10 (67%) experienced 
objective regressions including one ongoing complete regression at five months(3). In study 09-
C-0082, we have recently treated 12 patients with autologous PBL transduced with genes 
encoding an anti-CD19 CAR. Four of these patients have ongoing complete regressions from 4 
to 19 plus months and five additional patients have ongoing partial responses at 2 to 20 plus 
months. In both of these protocols only patients that have been heavily pre-treated with standard 
therapy have been admitted (early results published in (4, 5). 
Most toxicities observed in these studies were expected toxicities of the chemotherapy and 
aldesleukin administration. However, in 2 studies, we have observed serious adverse events 
related to the transduced cells. In 09-C-0041 (anti-Her2 CAR transduced PBL), the first patient, 
with metastatic colorectal cancer, was treated with 1010 autologous T cells transduced with the 
retrovirus encoding an anti-Her-2 CAR. This patient developed respiratory distress and died four 
days later. This toxicity was apparently due to a previously unrecognized ability of this CAR to 
recognize Her-2 expressed on lung epithelial cells although the exact explanation for the toxicity 
is not clear(6). In 09-C-0047 (anti-CEA TCR transduced PBL), all three treated patients 
experienced diarrhea, and colitis and one patient experienced an objective response of liver 
metastases(7). All gastrointestinal side effects resolved in these patients and the patients have 
normal bowel function. Grade 3 diarrhea lasting longer than 72 hours is considered a DLT per 
protocol and this event was observed in two of three patients enrolled in protocol 09-C-0047, 
meeting the criteria for stopping protocol accrual. 
1.2.2 MAGE-A3 as a target for cell transfer clinical studies 
Because of our success using the anti-NY-ESO-1 TCR we have explored targeting other cancer-
testes antigens. Cancer testis antigens (CTA) are proteins, which are normally only expressed in 
the placenta and in non-MHC expressing germ cells of testis, yet are aberrantly expressed in 
many tumors; thus CTA may represent ideal targets for tumor immunotherapy. More than 110 
CTA genes or gene families have been identified that are expressed in multiple tumor types (8-10). 
These proteins are being vigorously pursued as targets for therapeutic cancer vaccines, and TCR 
based adoptive immunotherapy (11-13). In theory, targeting T cells against tumor associated CT 
antigens might selectively eliminate tumor cells while avoiding toxicity to normal tissue. 


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Since the identification of the first human MAGE CT gene in 1991, the number of MAGE family 
genes has grown to over 25 members (14, 15). MAGE-A is a multigene family consisting of 12 
homologous genes, MAGE-A1 to A12 located at chromosome Xq28(10). Genomic clustering, 
restricted expression pattern and single exon open reading frames of the MAGE genes are 
consistent with the possibility that these genes evolved from retrotransposition and subsequent 
duplication(16). The precise function and biological role of MAGE proteins are not completely 
elucidated. However, members of MAGE-A, B and C proteins have been implicated in the 
suppression of p53-dependent apoptosis (17, 18) and MAGE-A3 has been attributed to mediate 
fibronectin-controlled tumor progression and metastasis(19). Expression of CTAs including 
MAGE genes in tumor cells has been attributed to global DNA demethylation and other 
mechanisms that normally silences these genes in somatic cells(20). MAGE-A3 is one of the more 
frequently expressed CT antigens in human tumors, including melanoma(21), non-small cell lung 
carcinoma(22), head and neck squamous cell carcinoma(23), pancreatic cancer(24), hepatocellular 
carcinoma(25), non-Hodgkin lymphoma(26), and multiple myeloma(27). The expression of MAGE-
A3 has been shown to be higher in more advanced stages of disease, and is associated with poor 
disease prognosis (28-30). Several antigenic peptides that bind to HLA class I or class II molecules 
on tumor cells have been reported (31-37). Because of its high expression in a wide array of tumor 
types, MAGE-A3 was chosen as the target for cancer immunotherapy. 
A high avidity Class I restricted TCR against MAGE-A3 was isolated using a transgenic mouse 
model that expressed the human HLA-A*0201 molecule. A retrovirus encoding the alpha and 
beta chains of this TCR that recognized the Class I restricted MAGE-A3 epitope (aa112-120) 
was constructed. Nine cancer patients were treated with adoptive cell therapy using autologous 
anti-MAGE-A3 T-cell receptor (TCR) engineered T cells. Five patients experienced clinical 
regression of their cancers including 2 on-going responders. However, postinfusion, 3 patients 
experienced mental status changes, and 2 patients lapsed into comas and subsequently died(38). 
The TCR used in this study recognized epitopes in MAGE-A3/A9/A12. Molecular assays of 
human brain samples using real-time quantitative-polymerase chain reaction, Nanostring 
quantitation, and deep-sequencing indicated that MAGE-A12 was expressed in human brain and 
our TCR recognized an epitope in MAGE-A12 that was similar to the MAGE-A3 epitope being 
targeted. This previously unrecognized expression of MAGE-A12 in human brain was possibly 
the initiating event of a TCR-mediated inflammatory response that resulted in neuronal cell 
destruction. There was no evidence of MAGE-A3 expressed in the brain. In a protocol using a 
TCR that recognized HLA-A1 Class I restricted epitope from MAGE-A3, cardiac toxicity was 
seen that was attributed to changes introduced into the binding regions of the(39) TCR, which 
added new specificities resulting in cardiac rejection. These changes were thought to be the 
cause of the off target toxicity. This study will use an unmodified human receptor and thus we do 
not expect to see the same off target toxicities. 
However, other MAGE-A3 epitopes may be good candidates for tumor immunotherapy because 
of the high percentage of tumors that express MAGE genes, (for example >60% of melanomas 
and >50% of NSCLC) and large numbers of patients can be eligible for treatment. Finally 
MAGE-A3 has not been found to be expressed in any normal tissue except testis and this may 
limit the risk of on-target toxicity to normal tissue. 


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1.2.3 Rationale for Targeting a Class II Restricted Epitope of MAGE-A3 recognized by CD4 T 
Cells 
The ability of CD8+ T cells that have been genetically modified to express TCRs that recognize 
HLA class I restricted antigens to mediate tumor regression has now been demonstrated in a 
series of clinical trials that target the melanocyte differentiation antigens (MDAs) MART-1 and 
gp100 (38, 40), as well as the cancer germ-line antigens MAGE-A3(2) and NY-ESO-1(3). 
Nevertheless, durable complete tumor regressions were observed in only a relatively small 
percentage of patients treated in these trials. Multiple factors may be responsible for the lack of 
durable responses seen in these patients, including insufficient persistence of transferred cells, 
heterogeneity of antigen and HLA class I expression, as well as the presence of host factors that 
suppress the response of adoptively transferred T cells. In addition, CD4+ T helper cells have 
been found to play a role in mediating anti-tumor immunity in murine tumor model systems, 
raising the possibility that lack of sufficient T cell help may impair the effectiveness of human 
therapies that have focused on the use of class I restricted TCRs. Tumor-reactive CD4+ T cells 
that recognize the murine leukemia virus epitopes (41, 42) as well as the TRP-1 melanocyte 
differentiation antigen(43) have been shown to be effective in mediating the regression of 
established murine tumors. Additional findings indicate that CD4+ T cells can enhance and 
maintain in vivo responses of tumor-reactive CD8+ T cells(44, 45). 
A variety of HLA class II restricted antigens expressed on human tumor cells have been 
identified, providing an opportunity to evaluate the effectiveness of CD4+ T cells in mediating 
tumor regression in patients. Multiple epitopes derived from MDAs, including MART-1(46), 
tyrosinase(47), gp100 (47-49) and TRP-1 (50, 51) have been shown to be recognized by human CD4+ 
tumor reactive T cells. In addition, HLA class II restricted epitopes derived from multiple cancer 
germ-line gene products, including MAGE-A3 (32, 34, 52, 53), MAGE-A1(54), NY-ESO-1 (55, 56), 
SSX-2(57) and SSX-4(58), have been identified by stimulating T cells in vitro with either 
recombinant proteins or peptides derived from these products. In the only clinical trial to 
specifically target an HLA class II restricted antigen, an autologous CD4+ T cell clone that 
recognized an HLA-DP*04-restricted epitope of NY-ESO-1 mediated tumor regression in a 
patient with melanoma(59). This finding provided an impetus to further investigate the role of 
CD4+ tumor reactive T cells in mediating tumor regression. 
We thus focused on the isolation of class II restricted TCRs directed against the MAGE-A3 
cancer germ-line antigens which is expressed in a variety of tumor histologies (60, 61). In previous 
studies aimed at identifying HLA class II restricted MAGE-A3 T cell epitopes, PBMC were 
stimulated with autologous dendritic cells (DCs) that had been pulsed with recombinant MAGE-
A3 protein(34). A single CD4+ T cell clone was identified in this study, clone 22, that recognized 
autologous EBV B cells that were either pulsed with recombinant MAGE-A3 protein or 
transduced with a recombinant retroviral construct encoding the MAGE-A3 protein. Further 
analysis lead to the identification of a peptide corresponding to amino acids 243-258 of the 
MAGE-A3 protein, KKLLTQHFVQENYLEY (MAGE-A3:243-258), that was recognized in 
association with two HLA class II allele DPβ1 alleles, *0401 and *0402. The DPβ1*0401 allele 
is expressed by approximately 70% of Caucasians, whereas the closely related DPβ1*0402 
allele, which differs from the 0401 allele by four amino acids, is expressed by approximately 
20% of Caucasians. Further analysis indicated that the minimal epitope recognized by clone 22 
corresponded to the 12-mer TQHFVQENYLEY (MAGE-A3:247-258). Targets cells expressing 
DPβ1*0401 that were either pulsed with the MAGE-A3:247-258 peptide or transduced with the 


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MAGE-A3 gene were strongly recognized by clone 22, whereas targets expressing the 
DPβ1*0402 allele were recognized to a lower but nevertheless significant extent. Subsequently, 
a patient clinical trial was carried out involving immunization of patients with MAGE-A3 
protein in combination with a group of HLA class I and class II restricted peptides that included 
the MAGE-A3:243-258 HLA-DPβ1*04 restricted peptide that were selected on the basis of the 
patients’ HLA haplotype(62). Although objective responses were not observed in this trial, 
subsequent in vitro studies were carried out to evaluate the phenotype and function of T cell 
clones from HLA-DPβ1*0401 or 0402+ patients that were isolated from the peripheral blood of 
immunized patients using a HLA-DP*04 tetramer containing the MAGE-A3:243-258 peptide. 
This procedure lead to the isolation of CD4+ MAGE-A3:243-258 reactive T effector clones that 
released IFN-γ and TNF-α following antigen stimulation, as well as T cell clones that appeared 
to represent CD4+ regulatory T cells (CD4+ Tregs) that recognized the same T cell epitope. 
These MAGE-A3 reactive CD4+ Treg clones failed to release effector cytokines following 
specific antigen stimulation, but were capable of suppressing effector T cell responses in 
response to stimulation with the MAGE-A3:243-258 peptide. 
Given the high prevalence of the HLA-DP*04 HLA class II allele in the patient population, as 
well as the high frequency of MAGE-A3+ tumors in patients with melanoma as well as other 
malignancies, we attempted to identify potent TCRs directed against the MAGE-A3:243-258 
HLA-DP*04 restricted epitope. Initially, we isolated the endogenous TCR alpha and beta chains 
expressed by two of the MAGE-A3:243-258-reactive effector CD4+ T cell clones, clones R12C9 
and 12C7, as well as a CD4+ Treg clone reactive with this epitope, each of which was 
characterized in the MAGE-A3 cancer vaccine trial study described above (kindly provided by 
Dr. Pierre van der Bruggen). Following the cloning of the TCR alpha and beta chains isolated 
from the three MAGE-A3 reactive clones they were then joined with the P2A self-cleaving 
peptide sequence and cloned in the MSGV1 retroviral vector. Three patient PBMC that had been 
stimulated with OKT3 were then transduced with transient retroviral supernatants on day two 
and enriched for CD4+ T cells on day seven, and assayed for their ability to recognize target cells 
on day 12. Analysis of the levels of transduction carried out by staining transduced cells with 
anti-Vβ22, Vβ6.7 and Vβ12 which detect the 6F9, R12C9 and 12A7 TCR (all kindly provided 
by Dr. Pierre van der Bruggen) indicated that the levels of transduction were similar, ranging 
between 25 and 35% (Figure 1a). The results of co-culture assays indicated that T cells 
transduced with either the 6F9, R12C9 or the 12C7 TCR strongly recognized HLA-DP*0401+ 
target cells pulsed with the MAGE-A3:243-258 peptide (Figure 1b). In contrast, CD4+ T cells 
transduced with the 6F9 but not the R12C9 or 12C7 TCRs recognized HLA DP*0401+ 293-
CIITA cells transfected with genes encoding MAGE-A3 or MAGE-A6, but not MAGE-A1 or 
A12 (Figure 1b). Comparison of amino acid sequences of the corresponding regions of the 
MAGE family members indicated that MAGE-A3 and MAGE-A6 only differed at one position 
(residue 249), whereas the other MAGE family members differed from MAGE-A3 at 2 (MAGE-
A12) or 3 (MAGE-A1) positions (Table 1a). In addition, CD4+ T cells transduced with the 6F9 
TCR but not the R12C9 or 12C7 TCRs recognized the MAGE-A3+/HLA-DP*0401+ melanoma 
cell line 1359 mel-CIITA but failed to recognize the MAGE-A3+/HLA-DP*0401- melanoma cell 
line 624 mel-CIITA. CD4+ T cells transduced with the R12C9 TCR failed to recognize either of 
the tested melanoma cell lines, whereas cells transduced with the 12C7 TCR weakly recognized 
624 mel-CIITA but not 1359 mel-CIITA cells (Figure 1b). Titration of the MAGE-A3:243-258 
peptide indicated that CD4+ T cells transduced with the 6F9 and R12C9 TCRs released 


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comparable levels of IFN-γ in response to targets pulsed with a minimum of between 0.001 and 
0.01 mg/mL of the MAGE-A3:243-258 peptide, whereas cells transduced with the 12C7 TCR 
appeared to release significantly less IFN-γ than those transduced with the other two TCRs 
(Figure 1c). These results indicated that while CD4+ T cells transduced with any of the three 
MAGE-A3 reactive TCRs 6F9, R12C9 and 12C7 TCR recognized peptide pulsed target cells, 
only cells transduced with the 6F9 TCR recognized transfected target cells as well as MAGE-
A3+ and HLA-DP*04+ tumor cells. 
The 6F9 TCR was further evaluated by determining the reactivity of CD4+ and CD8+ T cells 
separated from two patient PBMCs against a panel of tumor cell lines. Five melanoma cell lines 
that expressed MAGE-A3 and HLA-DP*0401, as well as the non-small cell lung carcinoma cell 
line H1299 NSCLC-CIITA were recognized by transduced CD4+ and CD8+ T cells, although 
CD4+ T cells secreted higher amounts of cytokine in response to tumor targets than transduced 
CD8+ T cells (Figure 2A). Prior studies have demonstrated higher reactivity of human TCRs in 
which sequences encoding the human TCR alpha and beta chain constant regions had been 
replaced with those encoding the corresponding murine constant regions(63). The effects of 
replacing the constant regions of the 6F9 TCR alpha and beta chains with murine sequences was 
then evaluated by constructing a retroviral vector encoding the murine constant regions, 
designated 6F9mc. Comparisons of responses of CD4+ T cells transduced with the normal 6F9 
TCR with those transduced with the 6F9mc TCR indicated that the murine constant regions 
resulted in between two and five-fold enhancement in the response of transduced T cells against 
the seven MAGE-A3+ and HLA-DP*0401+ targets that were evaluated (Figure 2B). In addition, 
the response of CD8+ T cells transduced with the 6F9mc were enhanced by between two and ten-
fold above those seen in cells transduced with the native 6F9 TCR. The responses of CD8+ T 
cells transduced with the 6F9mc were generally lower than CD4+ T cells transduced with this 
TCR, although comparable cytokine responses were observed in responses to some tumor targets 
(Figure 2C). 
In order to evaluate the fine specificity of antigen recognition mediated by cells transduced with 
the 6F9 and 6F9mc TCR, HLA-DP*0401+ target cells were pulsed with truncations of the 
MAGE-A3:243-258 peptide and related peptides from MAGE family members. Analysis of the 
response to truncated MAGE-A3 peptides from two cultures of transduced CD4+ PBMC 
indicated that the 11-mer peptide QHFVQENYLEY corresponding to amino acids 248-258 of 
the MAGE-A3 protein represented the minimal peptide that elicited a response comparable to 
that elicited by the MAGE-A3:243-258 peptide (Table 1B) that was nearly identical to the 
minimal epitope that was previously described but that lacked the amino terminal threonine 
residue present in that peptide(34). The MAGE-A3:243-258 peptide was predicted using an 
epitope prediction algorithm to possess a high affinity for HLA-DP*0401, and in addition, 
recognition of the truncated MAGE-A3 peptides appeared to correlate with T cell recognition 
(Table 1B). Significant recognition of the MAGE-A6:248-258 peptide that contained a single 
substitution of tyrosine for histidine at position 249, was seen but minimal reactivity was 
observed against additional members of the MAGE family of gene products that possessed 
between two and five differences from the MAGE-A3:248-258 peptide. A BLAST search of the 
NCBI database revealed that the most closely related peptide in the human genome was derived 
from the protein necdin. This peptide, which possessed five differences from the MAGE-
A3:248-258 peptide, was also not recognized by T cells transduced with the 6F9 or 6F9mc TCR. 
These findings indicate that the 6F9 TCR possesses a high degree of specificity for the MAGE-


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A3:248-258 peptide and suggest that T cells transduced with this TCR may possess little or no 
cross-reactivity with peptides derived from additional human proteins. 
To evaluate the anti-MAGE-A3 DP4 TCR at clinical scale, CliniMACs clinical-scale cell 
separation apparatus was used to deplete CD8+ lymphocytes from PBL. CD8-depleted PBL were 
stimulated for 2 days with OKT-3 antibody and then transduced twice by anti-MAGE-A3 DP4 
TCR retrovirus. Flow cytometric analysis of transduced PBL showed that 92.0% of CD4+ T cells 
were transduced with this TCR vector, detected by anti-mouse TCRβ antibody (Figure 3). In 
addition, 96.1% of transduced PBL were CD4+ T cells, with less than 0.1% CD8+ T cells. To 
assess the specific recognition of tumor cells, TCR-engineered CD4+ T cells were co-cultured 
with HLA-DP0401+ MAGE-A3+ target cells or MAGE-A3 peptide-pulsed cells. High levels of 
IFN-γ were observed from the co-culture wells with specific target cells but not negative controls 
(Figure 4A). Furthermore, TCR-engineered CD4+ T cells were rapidly expanded one time to 
achieve sufficient cell numbers for patient infusion, and the same biological assays were 
performed. Similar results were obtained, with 92.6% of transduced T cells and 99.2% of CD4+ 
T cells (Figure 3). Specific release of IFN-γ from TCR-engineered CD4+ T cells was also 
observed (Figure 4B). 
1.2.4 Safety Considerations 
Several safety concerns regarding the infusion of large numbers of retrovirally modified tumor 
reactive T-cells have been addressed in our previous clinical studies as described below. The 
non-myeloablative chemotherapy and the administration of high-dose aldesleukin have expected 
toxicities which are discussed in Section 6 of this study. The nonmyeloablative chemotherapy 
used in this protocol has been administered to over 300 patients and all have reconstituted their 
hematopoietic systems. 
In other protocols we have administered over 3 X 1011 TIL with widely heterogeneous 
reactivity including CD4, CD8, and NK cells without difficulty. We do not believe the 
transfer of these gene modified cells has a significant risk for malignant transformation in 
this patient population. While the risk of insertional mutagenesis is a known possibility 
using retroviral vectors, this has only been observed in the setting of infants treated for 
XSCID, WAS and X-CGD using retroviral vector-mediated gene transfer into CD34+ 
bone marrow cells. In the case of retroviral vector-mediated gene transfer into mature T-
cells, there has been no evidence of long-term toxicities associated with these procedures 
since the first NCI sponsored gene transfer study in 1989. Although continued follow-up 
of all gene therapy patients will be required, data suggest that the introduction of 
retroviral vectors transduced into mature T-cells is a safe procedure. While we believe the 
risk of insertional mutagenesis is extremely low, the 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 
a. Metastatic or locally advanced refractory/recurrent cancer that expresses MAGE-A3 
as assessed by one of the following methods: RT-PCR on tumor tissue defined as 
30,000 copies of MAGE-A3 per 106 GAPDH copies, or by immunohistochemistry of 


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resected tissue defined as 10% or greater of tumor cells being 2-3+ for MAGE-A3, or 
serum antibody reactive with MAGE-A3. Metastatic cancer diagnosis will be 
confirmed by the Laboratory of Pathology at the NCI. 
b. Patients must have previously received prior first line standard therapy (or effective 
salvage chemotherapy regimens) for their disease, if known to be effective for that 
disease, and have been either non-responders (progressive disease) or have recurred. 
c. Patients must be HLA-DP4 positive. 
d. Patients with 3 or fewer brain metastases that are less than 1 cm in diameter and 
asymptomatic are eligible. Lesions that have been treated with stereotactic radiosurgery 
must be clinically stable for 1 month after treatment for the patient to be eligible. 
Patients with surgically resected brain metastases are eligible. 
e. Greater than or equal to 18 years of age and less than or equal to age 70. 
f. Ability of subject to understand and the willingness to sign the Informed Consent 
Document 
g. Willing to sign a durable power of attorney 
h. Clinical performance status of ECOG 0 or 1 
i. Patients of both genders must be willing to practice birth control from the time of 
enrollment on this study and for up to four months after treatment. 
j. Serology:  
• 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 and more susceptible to its toxicities.)  
• Seronegative for hepatitis B antigen, and seronegative for hepatitis C antibody. If 
hepatitis C antibody test is positive, then patient must be tested for the presence 
of antigen by RT-PCR and be HCV RNA negative.  
k. Women of child-bearing potential must have a negative pregnancy test because of the 
potentially dangerous effects of the treatment on the fetus.  
l. Hematology 
• Absolute neutrophil count greater than 1000/mm3 without the support of 
filgrastim 
• WBC ≥ 3000/mm3 
• Platelet count ≥ 100,000/mm3 
• Hemoglobin > 8.0 g/dL 
m. Chemistry: 
• Serum ALT/AST ≤ to 2.5 times the upper limit of normal 
• Serum creatinine ≤ to 1.6 mg/dL 
• Total bilirubin ≤ to 1.5 mg/dL, except in patients with Gilbert’s Syndrome who 
must have a total bilirubin less than 3.0 mg/dL. 
n.More than four weeks must have elapsed since any prior systemic therapy at the time the 
patient receives the preparative regimen, and patients’ toxicities must have recovered to 
a grade 1 or less (except for toxicities such as alopecia or vitiligo). Patients must have 
progressing disease after prior treatment. Note: Patients who have previously received 
ipilimumab and have documented GI toxicity must have a normal colonoscopy with 
normal colonic biopsies. 
o.Subjects must be co-enrolled in protocol 03-C-0277. 


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2.1.2 Exclusion Criteria 
a. Women of child-bearing potential who are pregnant or breastfeeding because of the 
potentially dangerous effects of the treatment on the fetus or infant. 
b. Active systemic infections (e.g.: requiring anti-infective treatment), coagulation disorders 
or any other active major medical illnesses. 
c. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency 
Disease). 
d. Concurrent opportunistic infections (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 experimental treatment and more susceptible to 
its toxicities). 
e. Concurrent systemic steroid therapy. 
f. History of severe immediate hypersensitivity reaction to any of the agents used in this 
study. 
g. History of any cardiac events including coronary revascularization or ischemic 
symptoms. 
h. Documented LVEF of less than or equal to 45% testing is required in patients who are 
• ≥ 65 years’ old 
• Clinically significant atrial and or ventricular arrhythmias including but not 
limited to: atrial fibrillation, ventricular tachycardia, second or third degree heart 
block or have a history of ischemic heart disease, or chest pain. 
i. Documented FEV1 less than or equal to 60% predicted tested in patients with: 
• A prolonged history of cigarette smoking (20 pk/year of smoking within the past 
2 years). 
• Symptoms of respiratory dysfunction 
j. Patients who are receiving any other investigational agents. 
2.2 
SCREENING EVALUATION 
Note: Testing for screening evaluation is conducted under our companion protocol, 99-C-0128. 
2.2.1 Within 3 Months Prior to Enrollment 
a) HIV antibody titer and HBsAg determination, anti HCV (may be performed within 3 
months of chemotherapy start date). 
b) Confirmation of HLA-DP4 positivity. (Note: Testing is permitted to be conducted at any 
time prior to enrollment.) 
c) Confirmation of the diagnosis of cancer and MAGE A3 by the Laboratory of Pathology 
of the NCI. (Note: Testing is permitted to be conducted at any time prior to enrollment.) 
2.2.2 Within 8 Weeks Prior to Enrollment: 
a) Pulmonary Function testing for patients with a prolonged history of cigarette smoking (20 
pk/year of smoking within the past 2 years) or symptoms of respiratory dysfunction.  
b) Cardiac Evaluation (stress thallium, echocardiogram, MUGA etc.,) for patients who are 
greater than or equal to age 60, or have a history of ischemic heart disease, chest pain, or 
clinically significant atrial and/or ventricular arrhythmias including but not limited to: 
atrial fibrillation, ventricular tachycardia, heart block. Patients with a LEVF of less than 


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or equal to 45% will not be eligible. Patients under the age of 60 who have cardiac risk 
factors may also undergo cardiac evaluation as noted above (e.g., diabetes, hypertension, 
obesity).  
2.2.3 Within 4 Weeks Prior to Enrollment:  
a. Complete history and physical examination, including weight, vital signs, and noting in 
organ system involvement and any allergies/sensitivities to antibiotics. (Note: patient 
history may be obtained within 8 weeks.) 
b. Baseline imaging to determine the status of disease. This may include CT, MRI, PET, or 
photography.  
2.2.4 Within 14 Days Prior to Enrollment: 
a. Screening blood tests:  
• Chemistries: Creatinine, ALT/GPT, AST/GOT, and Total Bilirubin  
• CBC with differential and platelet count 
b. Urinalysis; urine culture, if indicated  
2.2.5  Within 7 Days Prior to Enrollment: 
• β-HCG pregnancy test (serum or urine) on all women of child-bearing potential 
• ECOG performance status of 0 or 1  
2.3 
 REGISTRATION AND TREATMENT ASSIGNMENT PROCEDURES 
2.3.1 Prior to registration for this protocol 
Patients will initially be registered on protocol 03-C-0277 (Cell Harvest and Preparation for 
Surgery Branch Adoptive Cell Therapy Protocols) prior to transduction of PBL cells (either fresh 
or cryopreserved samples), by the clinical fellow or research nurse.  
Once cells exceed the potency requirement and are projected to exceed the minimum number 
specified in the Certificate of Analysis (COA), patients will sign the consent document for this 
protocol. 
2.3.2 Registration procedure 
Authorized staff must register an eligible candidate with NCI Central Registration Office (CRO) 
within 24 hours of signing consent. A registration Eligibility Checklist from the web site 
(http://home.ccr.cancer.gov/intra/eligibility/welcome.htm) must be completed and sent via 
encrypted email to: 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. Verification of Registration will be 
forwarded electronically via e-mail to the research team. A recorder is available during non-
working hours.  
2.3.3 Treatment Assignment Procedures 
Cohorts 
Number Name 
Description 


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1 
MAGE-A3 (Phase 1) 
MAGE-A3-expressing metastatic or locally advanced 
refractory/recurrent cancer 
2 
Melanoma (Phase 2, 
Cohort I)  
Metastatic melanoma 
3 
Renal and Other (Phase 2, 
Cohort 2) 
Renal cancer and other types of metastatic cancer 
Arms 
Number Name 
Description 
1 
Phase 1 Experimental 
Therapy 
Non-myeloablative lymphodepleting preparative 
regimen of cyclophosphamide and fludarabine + Anti-
MAGE-A3-DP4 TCR PBL + high-dose aldesleukin 
2 
Phase 2 Experimental 
Therapy 
Non-myeloablative lymphodepleting preparative 
regimen of cyclophosphamide and fludarabine + Anti-
MAGE-A3-DP4 TCR PBL + high-dose aldesleukin 
Cohort and Arm Assignment 
• Subjects in Cohort 1 will be directly assigned to Arm 1. 
• Subjects in Cohort 2 will either be assigned to Arm 1 or Arm 2.  
3 
STUDY IMPLEMENTATION 
3.1 
STUDY DESIGN 
3.1.1 Performed on 03-C-0277 
PBMC will be obtained by leukapheresis (approximately 1 X 1010 cells). CD4+ cells will be 
enriched on Miltenyi columns as described in the IND and cultured in the presence of anti-CD3 
(OKT3) and aldesleukin in order to stimulate T-cell growth. Transduction is initiated by 
exposure of approximately 1 X 107 to 5 X 108 cells to supernatant containing the anti-MAGE-
DP4 retroviral vector. The CD4 enriched cells will then be further expanded in vitro prior to cell 
infusion. Successful TCR gene transfer will be determined by FACS analysis for the TCR 
protein and anti-tumor reactivity will be tested by cytokine release as measured on peptide 
pulsed DP4+ cells. Successful TCR gene transfer for each transduced PBL population will be 
defined as >10% TCR positive cells and for biological activity, gamma-interferon secretion must 
be at least 200pg/ml, and > 2 times background. 
3.1.2 Treatment Phase 
Patients will receive no other experimental agents while on this protocol. Patients will receive 
the standard NCI Surgery Branch non-myeloablative, lymphodepleting preparative regimen 
consisting of cyclophosphamide and fludarabine followed by IV infusion of anti-MAGE-A3-
DP4 TCR engineered PBL and aldesleukin. All patients will receive one course of treatment. The 
start date of the course will be the start date of the chemotherapy; the end date will be the day of 


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the first post-treatment evaluation. Patients may undergo a second treatment as described in 
Section 3.3. 
3.1.3 Dose Limiting Toxicity: 
Dose-limiting toxicity is defined as follows: All grade 3 and greater toxicities with the exception 
of: 
• Myelosuppression, defined as lymphopenia, neutropenia, decreased hemoglobin, and 
thrombocytopenia.  
• Aldesleukin expected toxicities as defined in Appendix 2 and Appendix 3. 
• Expected chemotherapy toxicities as defined in the Pharmaceutical Information section. 
• Immediate hypersensitivity reactions occurring within 2 hours of cell infusion (related to cell 
infusion) that are reversible to a grade 2 or less within 24 hours of cell administration with 
standard therapy. 
• Grade 3 Fever  
• Grade 3 Metabolic Laboratory abnormalities without significant clinical sequela that resolve 
to grade 2 within 7 days 
• Grade 3 autoimmunity, that resolves to less than or equal to a grade 2 autoimmune toxicity 
within 10 days 
Patients who develop toxicity due to study agents will be followed until toxicity resolves to 
grade 2 or less, regardless of disease progression. 
3.1.4 Phase I - Dose Escalation 
Initially, the protocol will enroll 1 patient in each dose level unless a patient experiences a dose 
limiting toxicity (DLT). The total number of anti-MAGE -A3-DP4 engineered PBL cells 
transferred for each dose level will be: 
 
Dose Level 
Dose 
Level 1 
1.0 x 107 cells 
Level 2 
3.0 x 107 cells 
Level 3 
1.0 x 108 cells 
Level 4 
3.0 x 108 cells 
Level 5 
1.0 x 109 cells 
Level 6 
3.0 x 109 cells 
Level 7 
1.0 x 1010 cells 
Level 8 
3.0 x 1010 cells 
Level 9 
1.0 x 1011 cells and up to 2.0 x 1011 cells 
 
In each dose level, if a patient experiences a DLT, a total of six patients will be treated at that 
dose to confirm that no greater than 1/6 patients have a DLT prior to proceeding to the next 
higher dose level. If a dose level with 2 or more DLTs in 3-6 patients has been identified, 
patients will be accrued at the next-lowest dose level, for a total of 6. If a dose limiting toxicity 
occurs in the first dose level, that dose level will be expanded to n=6 patients. If two DLTs occur 
in the first dose level, the study may be amended to treat patients at lower doses. If escalation is 


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20
permitted until Dose Level 9, it will be expanded to 6 patients unless it has 2 or more with DLTs 
before the 6th patient has been evaluated. 
The maximum tolerated cell dose is the highest dose at which < 1 of 6 patients experienced a 
DLT or the highest dose level studied if DLTs are not observed at any of the dose levels. 
A two-week safety assessment period following cell administration will be conducted between 
each dose level and before enrollment into the phase 2 portion of the protocol. 
3.1.5 Phase 2 Portion 
Similar to the Phase 1 portion, prior to receiving the engineered PBL cells, patients in the phase 
2 portion will receive a nonmyeloablative but lymphocyte depleting preparative regimen 
consisting of cyclophosphamide and fludarabine followed in one to four days by intravenous 
infusion of MAGE-A3-DP4 TCR transduced PBL. 
The phase 2 portion of the protocol will proceed using from 109 cells up to the MTD as 
determined in the phase 1 portion of the study. 
In the phase 2 portion of this study, patients will be entered into two cohorts based on histology: 
cohort 1 will include patients with metastatic melanoma; cohort 2 will include patients with renal 
cancer and other types of metastatic cancer. 
3.1.6 Protocol Stopping Rules 
New subject enrollment to the protocol will be temporarily halted if any of the following 
conditions are met, and discussions will be had with the FDA or NIH IRB regarding protocol 
revisions if applicable: 
• If 1 or more treatment related death occur due to the cell infusion, we will promptly 
discuss this with the NIH IRB and the FDA.  
• During the phase 1 portion of the study – if two or more patients develop a grade 3 or 
greater toxicity related to the cell product, with the exception of: 
•  Grade 3 metabolic laboratory abnormalities without significant clinical 
sequela that resolve to grade 2 or less within 7 days.  
• Grade 3 fever 
• If one of the first three patients (or 2 of the first 6 patients, or 3 of the first 9 patients, or 4 
of the first 12 patients) develop grade 3 autoimmunity, that cannot be resolved to less 
than or equal to a grade 2 autoimmune toxicity within 10 days, or any grade 4 or greater 
autoimmune toxicity. 
• During the phase 2 portion of the study - Once five or more patients have been enrolled, 
and if 20% or more patients cumulatively enrolled have developed a DLT, as described 
in Section 3.1.1. 
3.2 
DRUG ADMINISTRATION 
3.2.1 Preparative Regimen with Cyclophosphamide and Fludarabine 
(Starting on day -6, study medication start times for drugs given once daily should be given 
within 2 hours of the scheduled time. Chemotherapy infusions maybe slowed or delayed as 
medically indicated. Administration of diuretics, electrolyte replacement, and hydration and 
monitoring of electrolytes should all be performed as clinically indicated.) 


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21
DAYS -7 and -6  
Approximately 6 hours Prior to Cyclophosphamide 
Hydrate: Begin hydration with 0.9% Sodium Chloride Injection containing 10 meq/L of 
potassium chloride at 1.5 - 2.6 ml/kg/hr (starting approximately 6 hours’ pre-cyclophosphamide 
and continue hydration until 24 hours after last cyclophosphamide infusion). At any time during 
the preparative regimen, if urine output <1 ml/kg/hr or if body weight >2 kg over pre-
cyclophosphamide value, furosemide 10-20 mg IV maybe administered. The hydration rate will 
be capped at 250mL/hr. 
Approximately 1 hour pre-Cyclophosphamide 
Ondansetron (0.15 mg/kg/dose [rounded to the nearest even mg dose between 8 mg and 16 mg 
based on patient weight] IV every 8 hours X 3 days) will be given for nausea.  
Cyclophosphamide 60 mg/kg/day X 2 days IV in 250 ml D5W with Mesna 15 mg/kg/day X 2 
days over 1 hr. If patient is obese (BMI > 35) drug dosage will be calculated using practical 
weight as described in Appendix 1.  
A decreased dose of cyclophosphamide at 30mg/kg/day (x2 days) will be considered for patients 
who have a history of prolonged hematologic recovery from prior chemotherapy treatments. 
Immediately following the end of Cyclophosphamide 
Begin mesna infusion at 3 mg/kg/hour intravenously diluted in a suitable diluent (see 
pharmaceutical section) over 23 hours after each cyclophosphamide dose. If patient is obese 
(BMI > 35) drug dosage will be calculated using practical weight as described in Appendix 1.  
DAYS -7 to -3 
Fludarabine 25 mg/m2/day IVPB daily over 30 minutes for 5 days. If patient is obese (BMI > 35) 
drug dosage will be calculated using practical weight as described in Appendix 1. (The 
fludarabine will be started approximately 1-2 hours after the cyclophosphamide and mesna on 
Days -7 and -6)  
3.2.2 Cell Infusion and Aldesleukin Administration: 
The patient’s PBL is delivered to the patient care unit by a staff member from the Tumor 
Immunology Cell Processing Laboratory. Prior to infusion, the cell product identity label is 
double-checked by two authorized staff (MD or RN), an identification of the product and 
documentation of administration are entered in the patient’s chart, as is done for blood banking 
protocols. The cell dose will be given over 20-30 minutes or as clinically determined by an 
investigator for patient safety via non-filtered tubing, gently agitating the bag during infusion to 
prevent cell clumping. 
Aldesleukin will be administered at a dose of 720,000 IU/kg (based on total body weight) as an 
intravenous bolus over a 15-minute period approximately every 8 hours beginning within 24 
hours of cell infusion and continuing for up to 5 days (maximum 15 doses). Doses will be 
preferentially administered every eight hours; however, up to 24 hours may elapse between doses 
depending on patient tolerance. Aldesleukin dosing will be stopped if toxicities are not 
sufficiently recovered by supportive measures within 24 hours of the last dose of aldesleukin. 
Doses will be delayed or stopped if patients reach Grade 3 or 4 toxicity due to aldesleukin except 
for the reversible Grade 3 toxicities common to aldesleukin such as diarrhea, nausea, vomiting, 


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22
hypotension, skin changes, anorexia, mucositis, dysphagia, or constitutional symptoms and 
laboratory changes as detailed in Appendix 2. Toxicities will be managed as outlined in 
Appendix 3. In addition, dosing may be held or stopped at the discretion of the treating 
investigator. (Appendix 4 lists the toxicities seen in patients treated with aldesleukin at the NIH 
Clinical Center). Because confusion is a possible side effect of aldesleukin administration, a 
Durable Power of Attorney will be signed by the patient to identify a surrogate to make decisions 
if a patient becomes unable to make decisions. 
DAY 0 (one to four days after the last dose of fludarabine): 
 
• Cells will be infused intravenously (i.v.) on the Patient Care Unit over 20 to 30 minutes or 
as clinically determined by an investigator for patient safety via non-filtered tubing, gently 
agitating the bag during infusion to prevent cell clumping.  
• Aldesleukin as described above. 
DAY 0-4 (Day 0 is the day of cell infusion):  
• Beginning on day 1 or 2, filgrastim may be administered subcutaneously at a dose of 5 
mcg/kg/day (not to exceed 300 mcg/day). Filgrastim administration will continue daily 
until neutrophil count > 1.0 x109/L X 3 days or > 5.0 x109/L.  
• Aldesleukin as described above. 
3.2.3 Treatment Schedule 
1One to four days after the last dose of fludarabine 
2Initiate within approximately 24 hours after cell infusion 
3Continue until neutrophils count > 1X109/L for 3 consecutive days or > 5x109/L. 
Day 
-7 -6 
-5 
-4 
-3 
-2 
-1 
0 
1 
2 
3 
4 
Therapy 
 
 
 
 
 
 
 
 
 
 
 
 
Cyclophosphamide (60 mg/kg) 
X X 
 
 
 
 
 
 
 
 
 
 
Fludarabine (25 mg/m2) 
X 
X 
X 
X 
X 
 
 
 
 
 
 
 
Anti-MAGE-A3-DP4 TCR PBL 
 
 
 
 
 
 
 
X1 
 
 
 
 
Aldesleukin 
 
 
 
 
 
 
 
X2 
X 
X 
X 
X 
Filgrastim3 (5 mcg/kg/day) 
 
 
 
 
 
 
 
 
X 
X 
X 
X 
TMP/SMX4 
160mg/800 mg (example) 
 
 
 
 
 
 
 
X 
 
X 
 
X 
Fluconazole5 (400 mg po) 
 
 
 
 
 
 
 
X 
X 
X 
X 
X 
Valacyclovir po or Acyclovir IV6 
 
 
 
 
 
 
 
X 
X 
X 
X 
X 


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23
4The TMP/SMX schedule should be adjusted to QD three times per week (Monday, Wednesday, Friday) 
and continue for at least six months and until CD4 > 200 X 2, starting day 0 or within one week of 
anticipated lymphopenia 
5Continue until ANC > 1000/mm3 
6In patients positive for HSV or VZV continue until CD4 > 200 X 2 
3.3 
ON-STUDY EVALUATIONS 
Note: Please refer to section 5 for research evaluations. 
3.3.1 Within 14 days prior to starting the preparative regimen 
• Apheresis as indicated 
• Baseline blood test 
o Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, 
glucose, BUN), Hepatic Panel (alkaline phosphatase, AST, ALT, total 
bilirubin, direct bilirubin), Mineral Panel (albumin, calcium, magnesium, 
phosphorus), Uric Acid, Creatinine Kinase, Lactate Dehydrogenase, Protein, 
total  
o Complete Blood Count with differential 
o PT/PTT 
o TBNK 
o Thyroid Panel 
o Urinalysis 
• Anti CMV antibody titer, HSV and VZV serology, and EBV panel. (may be performed 
within 3 months of chemotherapy) 
• Chest x-ray 
• EKG 
3.3.2 During the preparative regimen: DAILY 
• Complete Blood Count with differential 
• Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN), 
Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct bilirubin), Mineral 
Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, Creatinine Kinase, Lactate 
Dehydrogenase, Protein, total 
• Urinalysis as needed 
• Daily weight as indicated 
• PT/PTT (every 3 days) 
3.3.3 After Cell Infusion: 
• Vital signs will be monitored hourly (+/- 15 minutes) for four hours and then routinely 
(every 4-6 hours) unless otherwise clinically indicated 
• Once total lymphocyte count is greater than 200/mm3, TBNK for peripheral blood CD4 
count will be drawn weekly (while the patient is hospitalized). Please refer to section 5 
for additional post cell infusion evaluations. 
3.3.4 During Hospitalization: 
• Every 1-2 days 


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o A review of systems and physical exam as clinically indicated 
o CBC with differential 
o Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, 
BUN), Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct 
bilirubin), Mineral Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, 
Creatinine Kinase, Lactate Dehydrogenase, Protein, total 
o Troponin levels on days 4 and 7 and as clinically indicated 
o Other tests will be performed as clinically indicated. 
o Vital signs and weight will be monitored as clinically indicated. 
3.4 
RETREATMENT  
Patients experiencing a partial or complete response may receive a second treatment when 
progression by RECIST criteria is documented after evaluation by the principal investigator. 
Patients will be retreated at the currently enrolling dose level but will not count towards 
establishing the MTD. Patients who develop grade 3 or grade 4 toxicity due to cell infusion will 
not be retreated. Patients must continue to meet the original eligibility criteria to be considered 
for retreatment. Toxicity related to cyclophosphamide, fludarabine, or aldeskeukin should be 
stable and resolved to less than grade 1 prior to retreatment. Retreatment benefits and risks will 
be carefully explained to the patient. A maximum of 1 retreatment course may occur. 
3.5 
POST TREATMENT EVALUATION  
• All patients will return to the NIH Clinical Center for their 1st evaluation for response 6 
weeks (+/- 2 weeks) following the administration of the cell product 
• Patients who have received multiple transfusions during the treatment phase or 
discharged with grade 3 or greater significant adverse events should be evaluated by 
referring physician and repeat labs as appropriate within 2 weeks of discharge. Labs 
drawn should be faxed to the study coordinator. 
3.5.1 Time period of evaluations 
Patients who experience stable disease, a partial response, or a complete response or have 
unresolved toxicities will be evaluated as noted below: 
• Week 12 (+/- 2 weeks)  
• Every 3 months (+/- 1 month) x3 
• Every 6 months (+/- 1 month) x 2 years 
• As per PI discretion for subsequent years 
Note: Patients may be seen more frequently as clinically indicated 
3.5.2  
At each scheduled evaluation, patients will undergo: 
• Physical examination, including weight and vital signs  
• Acute Care Panel (sodium, potassium, chloride, bicarbonate, creatinine, glucose, BUN), 
Hepatic Panel (alkaline phosphatase, AST, ALT, total bilirubin, direct bilirubin), Mineral 
Panel (albumin, calcium, magnesium, phosphorus), Uric Acid, Creatinine Kinase, Lactate 
Dehydrogenase, Protein, total  
• Complete blood count with differential 


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25
• PT/PTT 
• Urinalysis as needed 
• Thyroid panel as clinically indicated 
• TBNK, until CD4 > 200 X 2  
• Toxicity assessment, including a review of systems.  
• Imaging (CT, MRI, and/or PET) as performed at baseline. 
• A 5liter apheresis may be performed. If a patient is unable to undergo apheresis, 
approximately 96 ml of blood may be obtained at the first follow up visit. Subsequently, 
60 ml of blood will be obtained at follow up visits for at least 3 months. Peripheral blood 
mononuclear cells will be cryopreserved so that immunologic testing may be performed. 
This will be performed on 03-C-0277.  
• Detection of RCR and persistence of gene transduced cells (See Section 5.1.5) 
• Long-term follow up of patients receiving gene transfer: Physical examinations will be 
performed and documented annually for 5 years following cell infusion to evaluate long-
term safety. After 5 years, health status data will be obtained from surviving patients via 
telephone contact or mailed questionnaires. The long term follow up period for retroviral 
vectors is 15 years. This will be performed on 09-C-0161. 
NOTE: Patients who are unable or unwilling to return for follow up evaluations will be 
followed via phone or e-mail contacts. A request will be made to send laboratory, imaging 
and physician exam reports performed by their treating physician; and any outstanding 
toxicities will be reviewed with the patient.  
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 an evaluation 
safety visit approximately 6 weeks (± 2 weeks) following administration of the cell product (at 
the first follow-up evaluation).  
3.6.1 Criteria for removal from protocol therapy 
Patients will be taken off treatment (and followed until progression of disease) for the following: 
• Completion of protocol therapy 
• Participant requests to withdraw from active therapy 
• Investigator discretion 
• Positive pregnancy test 
3.6.2 Off Study Criteria 
Patients will be taken off study for the following: 
• Completion of study follow up period 
• The participant requests to be withdrawn from study 
• Progressive disease, unless the patient is eligible for a second treatment 
• Lost to follow-up 
• Death 
Note: Once a subject is taken off study, no further data can be collected. 


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26
Note: Patients who are taken off study for progressive disease or study closure maybe followed 
on Protocol 09-C-0161 “Follow up Protocol for Subjects Previously Enrolled in Surgery Branch 
Studies”.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 Updates 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. 
4 
CONCOMITANT MEDICATIONS/MEASURES 
4.1 
INFECTION PROPHYLAXIS: 
Note: Other anti-infective agents may be substituted at the discretion of the treating physician.  
4.1.1  Pneumocystis Jirovecii Pneumonia 
All patients will receive the fixed combination of trimethoprim and sulfamethoxazole 
(TMP/SMX) as double strength (DS) tab (DS tabs = TMP 160 mg/tab, and SMX 800 mg/tab) 
P.O. daily three times a week on non-consecutive days, beginning day 0 or within 1 week of 
lymphopenia.  
Dapsone (in G6PD sufficient patient), Atovaquone or Pentamidine may be substituted for 
TMP/SMX-DS in patients with sulfa allergies. 
4.1.2 Herpes or Varicella Zoster Virus Prophylaxis 
Patients with positive HSV or VZV serology will be given valacyclovir orally at a dose of 500 
mg daily starting the day of cell infusion, or acyclovir, 250 mg/m2 IV every 12 hrs if the patient 
is not able to take medication by mouth. Reversible renal insufficiency has been reported with IV 
but not oral acyclovir. Neurologic toxicity including delirium, tremors, coma, acute psychiatric 
disturbances, and abnormal EEGs has been reported with higher doses of acyclovir. Should this 
occur, a dosage adjustment will be made or the drug will be discontinued. Acyclovir will not be 
used concomitantly with other nucleoside analogs which interfere with DNA synthesis, e.g. 
ganciclovir. In renal disease, the dose is adjusted as per product labeling. 
Prophylaxis for Pneumocystis Varicella Zoster and Herpes will continue for 6 months’ post 
chemotherapy. If the CD4 count is less than 200 at 6 months’ post chemotherapy, prophylaxis 
will continue until the CD4 count is greater than 200 for 2 consecutive measures. 
4.1.3 Fungal Prophylaxis 
Patients will start Fluconazole 400 mg p.o. the day of cell infusion and continue until the 
absolute neutrophil count is greater than 1000/mm3. The drug may be given IV at a dose of 400 
mg in 0.9% sodium chloride USP daily in patients unable to take it orally. 
4.1.4 Empiric Antibiotics 
Patients will start on broad-spectrum antibiotics in accordance with current institutional 
guidelines for fever of 38.3°C once or two temperatures of 38.0°C or above at least one hour 
apart, AND an ANC <500/mm3. Infectious disease consultation will be obtained for all patients 
with unexplained fever or any infectious complications. 


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27
4.2 
BLOOD PRODUCT SUPPORT 
Using daily CBC’s as a guide, the patient will receive platelets and packed red blood cells 
(PRBC’s) as needed. Attempts will be made to keep Hb >8.0 gm/dl, and plts >10,000/mm3. All 
blood products will be irradiated. Leukocyte filters will be utilized for all blood and platelet 
transfusions to decrease sensitization to transfused WBC’s and decrease the risk of CMV 
infection. 
4.3 
OTHER CONCOMITANT MEDICATIONS TO CONTROL SIDE EFFECTS 
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 0.15 
mg/kg/dose IV every 8 hours will be administered for nausea and vomiting. Additional 
antiemetics will be administered as needed for nausea and vomiting uncontrolled by 
ondansetron. Antibiotic coverage for central venous catheters may be provided at the discretion 
of the investigator. 
5 
BIOSPECIMEN COLLECTION  
5.1 
CORRELATIVE STUDIES FOR RESEARCH:  
Blood and tissue are tracked at the patient level and can be linked to all protocols on which the 
patient has been enrolled. Samples will be used to support the specific objectives listed in the 
treatment protocol(s), e.g., immunologic monitoring, cytokine levels, persistence, as well as to 
support long term research efforts within the Surgery Branch and with collaborators as specified 
in our companion protocol, 03-C-0277 (Cell Harvest and Preparation for Surgery Branch 
Adoptive Cell Therapy Protocols). 
The amount of blood that may be drawn from adult patients for research purposes shall not 
exceed 10.5 mL/kg or 550 mL, whichever is smaller, over any eight-week period. 
5.2 
SAMPLES SENT TO DR. FIGG’S LAB 
• Venous blood samples will be collected in either a 4ml or an 8ml SST tube to be 
processed for serum and stored for future research. Record the date and exact time of 
draw on the tube. Blood tubes may be kept in the refrigerator until pickup.  
• For sample pickup, page 102-11964. 
• For immediate help, call 240-760-6180 (main blood processing core number) or, if no 
answer, 240-760-6190 (main clinical pharmacology lab number). 
• For questions regarding sample processing, contact Julie Barnes by e-mail or at 240-760-
6044. 
• The samples will be processed, barcoded, and stored in Dr. Figg’s lab until requested by 
the investigator. 
5.3 
SAMPLES SENT TO SURGERY BRANCH CELL PROCESSING LABORATORY 
• Venous blood samples will be collected in 8ml CPT tubes to be processed and stored for 
future research. Record the date and exact time of draw on the tube. Blood tubes are kept 
at room temperature until pickup. 


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28
• Samples will be pick-up by the research nurse or designee and transported to the SB Cell 
Processing Laboratory within 24 hours of blood draw.  
• The samples will be processed, barcoded, and stored in SB Cell Processing Laboratory. 
5.3.1 Prior to chemotherapy administration: 
• 5 CPT tubes (8 ml each) – SB’s lab 
• 1 SST tube (8 ml) – Figg’s lab 
• 1 SST tube (4ml) daily; starting the day chemotherapy begins – Figg’s lab 
5.3.2 Prior to cell infusion (one 8ml SST): 
• Baseline blood samples for cytokine analysis. – Figg’s lab 
5.3.3 Post cell infusion evaluations: 
Once total lymphocyte count is greater than 200/mm3, TBNK for peripheral blood CD4 count 
will be drawn and sent to the TIL lab on Monday, Wednesday and Friday x 5 days, then weekly 
(while the patient is hospitalized): 
• 5 CPT tubes (8 ml each) – SB’s lab 
• 1 SST tube (8 ml) – Figg’s lab 
5.4 
IMMUNOLOGICAL TESTING: 
• Apheresis may be performed prior to and 4-6 weeks (+/- 2 weeks) following the 
administration of the cell product. At other time points, patient peripheral blood 
lymphocytes (PBL) will be obtained from whole blood by purification using 
centrifugation on a Ficoll cushion. Aliquots of these PBMC will be 1) cryopreserved 
for immunological monitoring of cell function and 2) subjected to DNA and RNA 
extraction for PCR analysis of TCR and vector copy number estimation. 
• Lymphocytes will be tested directly and following in vitro culture. Direct 
immunological monitoring will consist of quantifying T cells reactive with MAGE-
A3/12 by FACS analysis using tetramer staining. Ex vivo immunological assays will 
consist of cytokine release by bulk PBL (+/- peptide stimulation) and by other 
experimental studies such as cytolysis if sufficient cells are available. If cell numbers 
are limiting, preference will be given to the direct analysis of immunological 
activity. Immunological assays will be standardized by the inclusion of 1) pre-
infusion PBMC and 2) an aliquot of the engineered PBL cryopreserved at the time of 
infusion. In general, differences of 2 to 3 fold in these assays are indicative of true 
biologic differences. Foxp3 levels will be analyzed by semiquantitative RT-PCR to 
evaluate for mRNA on PBL samples obtained prior to cell infusion and at the follow 
up time point. 
5.5 
MONITORING GENE THERAPY TRIALS: PERSISTENCE AND RCR: 
• Engineered cell survival. TCR and vector presence will be quantitated in PBMC 
samples using established PCR techniques. Immunological monitoring using both 
tetramer analysis and staining for the TCR will be used to augment PCR-based 
analysis. This will provide data to estimate the in vivo survival of lymphocytes 
derived from the infused cells.  


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29
• All patients will be co-enrolled on protocol 09-C-0161 “Follow up Protocol for 
Subjects Previously Enrolled in NCI Surgery Branch Studies”. Patients blood 
samples will be obtained and undergo analysis for detection of RCR by PCR prior to 
cell infusion and RCR PCR will be performed at 3 and 6 months, and at one year 
post cell administration. Blood samples will be archived annually thereafter if all 
previous testing has been negative with a brief clinical history. If a patient dies or 
develops neoplasms during this trial, efforts will be made to assay a biopsy sample 
for RCR. If any post-treatment samples are positive, further analysis of the RCR and 
more extensive patient follow-up will be undertaken, in consultation with the FDA. 
RCR PCR assays detect the GaLV envelope gene and are performed under contract 
by the Indiana University Vector Production Facility or other qualified testing 
facilities. The results of these tests are maintained by the contractor performing the 
RCR tests and by the Surgery Branch research team. 
5.6 
SAMPLE STORAGE, TRACKING AND DISPOSITION FOR SB CELL PROCESSING 
LABORATORY 
Blood and tissue collected during the course of this study will follow the Cell Tracking and 
Labeling System established by the Tumor Immunology Cell Processing Laboratory. The Cell 
Tracking and Labeling System is designed to unambiguously ensure that patient/data verification 
is consistent. The patients' cell samples (blood or tissue) are tracked by distinct identification 
labels that include a unique patient identifier and date of specimen collection. Cryopreserved 
blood and tissue samples also bear the date the sample was frozen. All cryopreserved samples are 
tracked for freezer location and storage criteria. All samples are stored in monitored 
freezers/refrigerators in 3NW Surgery Branch Laboratories at specified temperatures with alarm 
systems in place. Serum samples will be sent to the Blood Processing Core (BPC) for storage. 
Samples will be barcoded and stored on site or offsite at NCI Frederick Central Repository 
Services in Frederick, MD. All samples collected (blood or tissue) are entered into a central 
computer database with identification and storage location, and this database is backed up every 
night. 
If, at any time, a patient withdraws from the study and does not wish for their existing samples to 
be utilized, the individual must provide a written request. Following receipt of this request, the 
samples will be destroyed (or returned to the patient, if so requested), and reported as such to the 
IRB. Any samples lost (in transit or by a researcher) or destroyed due to unknown sample 
integrity (i.e. broken freezer allows for extensive sample thawing, etc.) will be reported as such 
to the IRB. 
Note: Blood and tissue collected during the course of this study will be stored, tracked and 
disposed of as specified in our companion protocol 03-C-0277, (Cell Harvest and Preparation for 
Surgery Branch Adoptive Cell Therapy Protocols). 
5.7 
SAMPLE STORAGE, TRACKING AND DISPOSITION FOR DR. FIGG’S LAB 
5.7.1 Sample Data Collection 
All samples sent to the Blood Processing Core (BPC) will be barcoded, with data entered and 
stored in the LABrador (aka LabSamples) utilized by the BPC, and data will be updated to the 
Surgery Branch central computer database weekly. This is a secure program, with access to 
LABrador limited to defined Figg lab personnel, who are issued individual user accounts. 


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I nstallati o n of L A Bra d or is li mite d t o c o m p uters s pecifie d b y Dr. Fi g g. T hese c o m p uters all ha ve 
a pass w or d restricte d l o gi n scree n. All Fi g g la b pers o n nel wit h access t o patie nt i nf or mati o n 
a n n uall y c o m plete t he NI H o nli ne Pr otecti o n of H u ma n S u bjects c o urs e.  
L A Bra d or creates a u ni q ue barc o de I D f or e ver y s a m ple a n d sa m ple b o x, w hic h ca n n ot be trace d 
bac k t o patie nts wit h o ut L A Bra d or access. T he data rec or de d f or eac h sa m ple i ncl u des t he 
patie nt I D, na me, trial na me/ pr ot oc ol n u m ber, ti me dra w n, c ycle ti me p oi nt, d os e, material t y pe, 
as well as b o x a n d freezer l ocati o n. Patie nt de m o gra p hics ass ociate d wit h t he cli nical ce nter 
patie nt n u m ber are pr o vi de d i n t he s yste m. F or eac h s a m ple, t here are n otes ass ociate d wit h t he 
pr ocessi n g met h o d ( dela y i n s a m ple pr ocessi n g, st ora ge c o n diti o ns o n t he war d, etc.). 
5. 7. 2  Sa m ple St ora ge a n d Destr ucti o n 
B arc o de d sa m ples are st ore d i n barc o de d b o xes i n a l oc ke d freezer at eit her - 2 0 or - 8 0C 
acc or di n g t o sta bilit y re q uire me nts. T hese freezers  are l ocate d o nsite i n t he B P C a n d offsite at 
N CI Fre deric k Ce ntral Re p osit or y Ser vices i n Fre deric k, M D. Visit ors t o t he la b orat or y are 
re q uire d t o be acc o m pa ni e d b y la b orat or y staff at all ti mes.  
A ccess t o st ore d cli nical sa m ples is restricte d. Sa m ples will be st ore d u ntil re q ueste d b y a 
researc her na me d o n t he pr ot oc ol. All re q uests are m o nit ore d a n d trac ke d i n L A Bra d or. All 
researc hers are re q uire d t o si g n a f or m stati n g t hat t he sa m ples are o nl y t o b e us e d f or researc h 
p ur p os es ass ociate d wit h t his trial (as per t he I R B a p pr o ve d pr ot oc ol) a n d t hat a n y u n us e d 
sa m ples m ust be ret ur ne d t o t he B P C. It is t he res p o nsi bilit y of t he N CI Pri nci pal I n vesti gat or t o 
e ns ure t hat t he sa m ples re q ueste d are bei n g use d i n a ma n ner c o nsiste nt wit h I R B a p pr o val.  
F oll o wi n g c o m pleti o n of t his st u d y, s a m ples will re mai n i n st ora ge as detaile d a b o ve. A ccess t o 
t hese s a m ples will o nl y b e gra nte d f oll o wi n g I R B a p pr o val of a n a d diti o nal pr ot oc ol, gra nti n g 
t he ri g hts t o use t he material.  
If, at a n y ti me, a patie nt wit h dra ws fr o m t he st u d y a n d d oes n ot wis h f or t heir e xisti n g sa m ples t o 
be utilize d, t he i n di vi d ual m ust pr o vi de a writte n re q uest. F oll o wi n g recei pt of t his re q uest, t he 
sa m ples will be destr o ye d ( or ret ur ne d t o t he patie nt, if s o re q ueste d), a n d re p orte d as s uc h t o t he 
I R B. A n y s a m ples l ost (i n tra nsit or b y a res earc her) or destr o ye d d ue t o u n k n o w n s a m ple 
i nte grit y (i.e. br o ke n freezer all o ws f or e xte nsi ve sa m ple t ha wi n g, etc.) will be re p orte d as s uc h 
t o t he I R B. 
Sa m ple barc o des are li n ke d t o patie nt de m o gra p hics a n d li mite d cli nical i nf or mati o n. T his 
i nf or mati o n will o nl y be pr o vi de d t o i n vesti gat ors liste d o n t his pr ot oc ol, vi a re gistere d use of t he 
L A Bra d or. It is critical t hat t he s a m ple re mai ns li n ke d t o patie nt i nf or mati o n s uc h as race, a ge, 
dates of dia g n osis a n d deat h, a n d hist ol o gical i nf or mati o n a b o ut t he t u m or, i n or der t o c orrelate 
ge n ot y pe wit h t hes e varia bles. 
N ote:  Bl o o d a n d tiss ue c ollecte d d uri n g t he c o urse of t his st u d y will be st ore d, trac ke d a n d 
dis p ose d of as s pecifie d i n o ur c o m pa ni o n pr ot oc ol 0 3- C- 0 2 7 7, ( Cell H ar vest a n d Pre parati o n f or 
S ur ger y Bra nc h A d o pti ve Cell T hera p y Pr ot oc ols).   
6  
D A T A C O L L E C TI O N A N D E V A L U A TI O N  
6. 1  
D A T A C O L L E C TI O N  
T he PI will be res p o nsi ble f or o vers eei n g e ntr y of data i nt o a n i n -h o us e pass w or d pr otecte d 
electr o nic s yste m ( C 3 D) a n d e ns uri n g data acc urac y, c o nsiste nc y a n d ti meli ness. T he pri nci pal 
i n vesti gat or, ass ociate i n vesti gat ors/researc h n urses a n d/ or a c o ntracte d data ma na ger will assist 


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31
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. Data will be entered into the NCI CCR 
C3D database. 
All adverse events (AEs), 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 AEs until their first week 6 follow-up evaluation or until off-
study, whichever comes first. 
An abnormal laboratory value will be recorded in the database as an AE only if the laboratory 
abnormality is characterized by any of the following: 
• Results in discontinuation from the study 
• Is associated with clinical signs or symptoms  
• Requires treatment or any other therapeutic intervention 
• Is associated with death or another serious adverse event, including hospitalization.  
• Is judged by the Investigator to be of significant clinical impact 
• If any abnormal laboratory result is considered clinically significant, the investigator will 
provide details about the action taken with respect to the test drug and about the patient’s 
outcome. 
All AEs must be recorded on the AE case report form unless otherwise noted below in Section 
6.1.1. 
End of study procedures: Data will be stored according to HHS and 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 Recording: 
Patients will be receiving multiple agents which include commercially available agents 
(fludarabine, cyclophosphamide and supportive medications) in combination with the 
investigational agents; therefore, grade 1 events not related to the cell product will not be 
reported/recorded. 
6.2 
DATA SHARING PLAN 
6.2.1 Human Data Sharing Plan 
De-identified human data generated for use in future and ongoing research will be shared 
through a NIH-funded or approved repository (ClinicalTrials.gov) and BTRIS. At the completion 
of data analysis, data will be submitted to ClinicalTrials.gov either before publication or at the 
time of publication or shortly thereafter. Data may also be used to support long term research 
efforts within the Surgery Branch and de-identified data may also be shared with collaborators as 
specified in our companion protocol, 03-C-0277 (Cell Harvest and Preparation for Surgery 
Branch Adoptive Cell Therapy Protocols). 


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6.2.2 Genomic Data Sharing Plan 
The NIH Genomic Data Sharing Policy does not apply to this study. 
6.3 
RESPONSE CRITERIA 
For the purposes of this study, patients should be re-evaluated for response at 6 and 12 weeks 
(+/- 2 weeks), then every 3 months (+/- 1 month) x3, then every 6 months’ (+/- 1 month) x 2 
years. In addition to a baseline scan, confirmatory scans should also be obtained approximately 4 
(not less than 4) weeks following initial documentation of objective response. 
Clinical Response will be determined using the Response Evaluation Criteria in Solid Tumors 
(RECIST) guideline (version 1.0).  
6.3.1  Definitions 
Evaluable for toxicity: All patients will be evaluable for toxicity from the time of their first 
treatment with Cyclophosphamide. 
Evaluable for objective response: Only those patients who have measurable disease present at 
baseline, have received at least one cycle of therapy, and have had their disease re-evaluated will 
be considered evaluable for response. These patients will have their response classified 
according to the definitions stated below. (Note: Patients who exhibit objective disease 
progression prior to the end of cycle 1 will also be considered evaluable.) 
Evaluable Non-Target Disease Response: Patients who have lesions present at baseline that are 
evaluable but do not meet the definitions of measurable disease, have received at least one cycle 
of therapy, and have had their disease re-evaluated will be considered evaluable for non-target 
disease. The response assessment is based on the presence, absence, or unequivocal progression 
of the lesions. 
6.3.2 Disease Parameters 
Measurable disease: Measurable lesions are defined as those that can be accurately measured in at 
least one dimension (longest diameter to be recorded) as:  
• By chest x-ray: >20 mm;  
• By CT scan:  
o Scan slice thickness 5 mm or under as >10 mm with CT scan 
o Scan slice thickness >5 mm: double the slice thickness  
• With calipers on clinical exam: >10 mm.  
All tumor measurements must be recorded in millimeters (or decimal fractions of centimeters). 
Malignant lymph nodes: To be considered pathologically enlarged and measurable, a lymph 
node must be >15 mm in short axis when assessed by CT scan (CT scan slice thickness 
recommended to be no greater than 5 mm). At baseline and in follow-up, only the short axis will 
be measured and followed. 
Non-measurable disease: All other lesions (or sites of disease), including small lesions (longest 
diameter <10 mm or pathological lymph nodes with ≥10 to <15 mm short axis), are considered 
non-measurable disease. Bone lesions, leptomeningeal disease, ascites, pleural/pericardial 


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33
effusions, lymphangitis cutis/pulmonitis, inflammatory breast disease, and abdominal masses 
(not followed by CT or MRI), are considered as non-measurable. 
Target lesions: All measurable lesions up to a maximum of 2 lesions per organ and 5 lesions in 
total, representative of all involved organs, should be identified as target lesions and recorded 
and measured at baseline. Target lesions should be selected on the basis of their size (lesions 
with the longest diameter), be representative of all involved organs, but in addition should be 
those that lend themselves to reproducible repeated measurements. It may be the case that, on 
occasion, the largest lesion does not lend itself to reproducible measurement in which 
circumstance the next largest lesion which can be measured reproducibly should be selected. A 
sum of the diameters (longest for non-nodal lesions, short axis for nodal lesions) for all target 
lesions will be calculated and reported as the baseline sum diameters. If lymph nodes are to be 
included in the sum, then only the short axis is added into the sum. The baseline sum diameters 
will be used as reference to further characterize any objective tumor regression in the measurable 
dimension of the disease. 
Non-target lesions: All other lesions (or sites of disease) including any measurable lesions over 
and above the 5 target lesions should 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 should be noted throughout follow-up. 
6.3.3 Methods for Evaluation of Measurable Disease 
All measurements should be taken and recorded in metric notation using a ruler or calipers. All 
baseline evaluations should be performed as closely as possible to the beginning of treatment and 
never more than 4 weeks before the beginning of the treatment. 
The same method of assessment and the same technique should be used to characterize each 
identified and reported lesion at baseline and during follow-up. Imaging-based evaluation is 
preferred to evaluation by clinical examination unless the lesion(s) being followed cannot be 
imaged but are assessable by clinical exam. 
Clinical lesions: Clinical lesions will only be considered measurable when they are superficial 
(e.g., skin nodules and palpable lymph nodes) and ≥10 mm diameter as assessed using calipers 
(e.g., skin nodules). In the case of skin lesions, documentation by color photography, including a 
ruler to estimate the size of the lesion, is recommended.  
Conventional CT and MRI: This guideline has defined measurability of lesions on CT scan based 
on the assumption that CT slice thickness is 5 mm or less. If CT scans have slice thickness 
greater than 5 mm, the minimum size for a measurable lesion should be twice the slice thickness.  
MRI is also acceptable in certain situations (e.g. for body scans). Ideally, the same type of 
scanner should be used and the image acquisition protocol should be followed as closely as 
possible to prior scans. Body scans should be performed with breath-hold scanning techniques, if 
possible. 
FDG-PET: While FDG-PET response assessments need additional study, it is sometimes 
reasonable to incorporate the use of FDG-PET scanning to complement CT scanning in 
assessment of progression (particularly possible 'new' disease). New lesions on the basis of FDG-
PET imaging can be identified according to the following algorithm:  


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a) Negative FDG-PET at baseline, with a positive FDG-PET at follow-up is a sign of PD 
based on a new lesion. 
b) No FDG-PET at baseline and a positive FDG-PET at follow-up: If the positive FDG-PET 
at follow-up corresponds to a new site of disease confirmed by CT, this is PD. If the 
positive FDG-PET at follow-up is not confirmed as a new site of disease on CT, 
additional follow-up CT scans are needed to determine if there is truly progression 
occurring at that site (if so, the date of PD will be the date of the initial abnormal FDG-
PET scan). If the positive FDG-PET at follow-up corresponds to a pre-existing site of 
disease on CT that is not progressing on the basis of the anatomic images, this is not PD. 
c) FDG-PET may be used to upgrade a response to a CR in a manner similar to a biopsy in 
cases where a residual radiographic abnormality is thought to represent fibrosis or 
scarring. The use of FDG-PET in this circumstance should be prospectively described in 
the protocol and supported by disease-specific medical literature for the indication. 
However, it must be acknowledged that both approaches may lead to false positive CR 
due to limitations of FDG-PET and biopsy resolution/sensitivity. 
Note: A ‘positive’ FDG-PET scan lesion means one which is FDG avid with an uptake greater 
than twice that of the surrounding tissue on the attenuation corrected image. 
6.3.4 Response Criteria 
6.3.4.1 Evaluation of target lesions1  
• 
Complete Response (CR): Disappearance of all target lesions 
• 
Partial Response (PR): At least a 30% decrease in the sum of the longest diameter 
(LD) of target lesions taking as reference the baseline sum LD. 
• 
Progression (PD): At least a 20% increase in the sum of LD of target lesions 
taking as reference the smallest sum LD recorded since the treatment started or 
the appearance of one or more new lesions. 
• 
Stable Disease (SD): Neither sufficient shrinkage to qualify for PR nor sufficient 
increase to qualify for PD taking as references the smallest sum LD. 
6.3.4.2 Evaluation of non-target lesions2  
• 
Complete Response (CR): Disappearance of all non-target lesions and 
normalization of tumor marker level. 
• 
Non-Complete Response: Persistence of one or more non-target lesions 
• 
Progression (PD): Appearance of one or more new lesions. Unequivocal 
progression of existing non-target lesions 
----------------------------------------------------------------------------------- 
1 
All measurable lesions up to a maximum of 10 lesions representative of all involved organs should be identified as target 
lesions and recorded and measured at baseline. Target lesions should be selected on the basis of their size (lesions with 
the longest diameter) and their suitability for accurate repetitive measurements (either by imaging techniques or clinically). 
A sum of the longest diameter (LD) for all target lesions will be calculated and reported as the baseline sum LD. The 
baseline sum LD will be used as reference to further characterize the objective tumor response of the measurable 
dimension of the disease. 
2 
All other lesions (or sites of disease) should be identified as non-target lesions and should also be recorded at baseline. 
Measurements are not required, and these lesions should be followed as “present” or “absent.” 
 


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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). The patient's best 
response assignment will depend on the achievement of both measurement and 
confirmation criteria. 
 
Target Lesions 
Non-Target Lesions 
New Lesions 
Overall Response 
CR 
CR 
No 
CR 
CR 
Non-CR/Non-PD 
No 
PR 
PR 
Non-PD 
No 
PR 
SD 
Non-PD 
No 
SD 
PD 
Any 
Yes or No 
PD 
Any 
PD 
Yes or No 
PD 
Any 
Any 
Yes 
PD 
6.3.5 Confirmatory Measurement/Duration of Response 
Confirmation 
To be assigned a status of PR or CR, changes in tumor measurements must be confirmed by 
repeat studies that should be performed at least 4 weeks after the criteria for response are first 
met. In the case of SD, follow-up measurements must have met the SD criteria at least once after 
study entry at a minimum interval of 6-8 weeks. 
Duration of Overall Response 
The duration of overall response is measured from the time measurement criteria are met for 
CR/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). 
The duration of overall complete response is measured from the time measurement criteria are 
first met for CR until the first date that recurrent disease is objectively documented. 
Duration of Stable Disease 
Stable 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.  
6.4 
TOXICITY CRITERIA 
The following adverse event management guidelines are intended to ensure the safety of each 
patient while on the study. The descriptions and grading scales found in the revised NCI 
Common Terminology Criteria for Adverse Events (CTCAE) version 4.0 will be utilized for AE 
reporting. 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). 


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7 
SAFETY REPORTING REQUIREMENTS/DATA AND SAFETY MONITORING 
PLAN 
7.1 
DEFINITIONS 
7.1.1 Adverse Event 
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 .  
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: 
• 
Death 
• 
A life-threatening adverse drug experience 
• 
Inpatient hospitalization or prolongation of existing hospitalization 
• 
A persistent or significant disability/incapacity or substantial disruption of the ability to 
conduct normal life functions.  
• 
A congenital anomaly/birth defect. 
• 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse drug experience 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. 


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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.  
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: 
•  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 
• Is related or possibly related to participation in the research; AND 
• Suggest 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 (CD) REPORTING 
7.2.1 NCI-IRB and NCI CD 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: 
• All deaths, except deaths due to progressive disease 
• All Protocol Deviations 
• All Unanticipated Problems 
• 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 
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: 
• All Grade 2 unexpected events that are possibly, probably or definitely related to the 
research;  
• All Grade 3 and 4 events that are possibly, probably or definitely related to the 
research;  
• All Grade 5 events regardless of attribution;  


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• 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 
From the time the subject receives the investigational agent/intervention to the time of the first 
follow-up evaluation (6 weeks (± 2 weeks) following the administration of the cell product), the 
investigator must immediately report to the sponsor, using the mandatory MedWatch Form FDA 
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 after the first follow-up evaluation, only those events that have an attribution of 
at least possibly related to the agent/intervention will be reported. 
Required timing for reporting per the above guidelines: 
• 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 Waivier of expedited reporting to CCR 
The investigators are requesting a waiver from reporting specific events in an expedited manner 
to the CCR. Patients will be receiving commercially available agents, such as fludarabine, 
cyclophosphamide, and aldesleukin. The majority of toxicities observed on Surgery Branch 
adoptive cell therapy protocols are expected toxicities of the non-myeloablative chemotherapy 
regimen or IL-2 and occur in approximately 95% of the patients enrolled, therefore, we are 
requesting a waiver from reporting the following events in an expedited manner to the CCR;  
• Grade 3 or greater myelosuppression, defined as lymphopenia, neutropenia, decreased 
hemoglobin, and thrombocytopenia. 
• Grade 3 or greater nausea, vomiting, mucositis - oral, anorexia, diarrhea, fever, chills, 
fatigue, and rash maculo-papular. 
• Grade 3 hypoxia, dyspnea, hematuria, hypotension, sinus tachycardia, urine output 
decreased, confusion, infections, and febrile neutropenia. 
The PI will submit a summary table of all grade 3-5 events, whether or not considered related to 
the product, every 6 months. The report shall include the number of patients treated in the 
timeframe, the number of events per AE term per grade which occurred in the 6-month 
timeframe and in total since the start of the study, attribution, and type/category of serious. 
Reports will be submitted to the Center for Cancer Research (CCR) at: 
CCRsafety@mail.nih.gov. 


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7.3.2 Reporting Pregnancy  
7.3.2.1 Maternal exposure 
If a patient becomes pregnant during the first four months following treatment the pregnancy 
should be 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 a SAE 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 
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 one 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. 
7.3.2.2 Paternal exposure 
Male patients should refrain from fathering a child or donating sperm during the study and for 
120 days after the last dose of aldesleukin. 
Pregnancy of the patient’s partner is not considered to be an AE. However, the outcome of all 
pregnancies (spontaneous miscarriage, elective termination, ectopic pregnancy, normal birth, or 
congenital abnormality) occurring from the date of the first dose until 120 days after the last dose 
should, if possible, be followed up and documented. 
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 Anti-MAGE-A3-DP4 TCR PBL 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 Anti-MAGE-A3-DP4 TCR 
PBL, but are not fatal or life-threatening, must 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. 


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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 for each trial:  
• the title and purpose of the trial  
• clinical site 
• the Principal Investigator 
• clinical protocol identifiers  
• participant population (such as disease indication and general age group, e.g., adult or 
pediatric);  
• 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  
• the status of the trial, e.g., open to accrual of subjects, closed but data collection ongoing, 
or fully completed,  
• 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: 
• a narrative or tabular summary showing the most frequent and most serious adverse 
experiences by body system 
• a summary of all serious adverse events submitted during the past year 
• 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 
• if any deaths have occurred, the number of participants who died during participation in 
the investigation and causes of death 
• 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 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 or a lead 
associate investigator. Adverse events will be reported as required above. Any safety concerns, 


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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 subject’s protection. This is done through independent verification of study 
data with source documentation focusing on: 
• Informed consent process 
• Eligibility confirmation 
• Drug administration and accountability 
• Adverse events monitoring 
• Response assessment. 
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  
Following an initial portion of the study in which the safety of cells used in this protocol will be 
determined, the primary objective of this trial is to determine whether the combination of high 
dose aldesleukin, lymphocyte-depleting chemotherapy, and an infusion of anti-MAGE-A3-DP4-
TCR-gene engineered lymphocytes is able to be associated with a modest fraction of patients that 
can experience a clinical response (PR +CR) to therapy. A secondary objective is to have 
sufficient patients in order to do exploratory evaluations of survival of cells.  
The study will begin by evaluating the safety of escalating doses of cells from 107 to 1011. Once 
a safe dose has been confirmed, patients will be enrolled into the phase 2 portion of the trial 
using the cell dose found to be safe in the first phase of this protocol. 


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Following the determination of a safe cell dose, the following will describe the phase 2 portion 
of the study. 
Patients who express MAGE-A3-DP4 will be enrolled into individual strata depending on their 
specific histology. This stratification is being used to separate patients who have been shown 
historically to be aldesleukin sensitive (melanoma) from those who have not. In each case, the 
MAGE-A3-DP4 expression is expected to be so dominant in regulating potential for response 
that differences in clinical response according to histology will not be expected but will be 
evaluated in an exploratory manner to help assess this, in a pilot sense. 
For each of the 2 strata, the study will be conducted using a phase 2 optimal design (Simon R, 
Controlled Clinical Trials 10:1-10, 1989). For both strata, the objective will be to determine if 
the combination of high dose aldesleukin, lymphocyte depleting chemotherapy, and anti-MAGE-
A3-DP4-TCR-gene engineered lymphocytes is able to be associated with a clinical response rate 
that can rule out 5% (p0=0.05) in favor of a modest 20% PR + CR rate (p1=0.20). 
In patients in each of the two strata (melanoma vs. other histologies or renal carcinoma), the 
following design will be used. For each strata, with alpha=0.05 (5% probability of accepting a 
poor therapy) and beta=0.10 (10% probability of rejecting a good therapy,), initially 21 evaluable 
patients will be enrolled. If 0 or 1 of the 21 patients experiences a clinical response, then no 
further patients will be enrolled. If 2 or more of the first 21 evaluable patients enrolled have a 
clinical response, then accrual will continue until a total of 41 evaluable patients have been 
enrolled. As it may take several weeks to determine if a patient has experienced a clinical 
response, a temporary pause of up to 6 months in the accrual to the trial may be necessary to 
ensure that enrollment to the second stage is warranted. If 2 to 4 of the 41 have a clinical 
response, then this will be considered inadequate for further investigation. If 5 or more of 41 
patients have a clinical response, then this will indicate that this strategy provides a new 
approach that may be worthy of further consideration. Under the null hypothesis (5% response 
rate), the probability of early termination is 72%. 
Further, to help ensure that maldistribution of patients who are either particularly responsive or 
unresponsive in the first stage does not materially interfere with the intended use of the two-stage 
design, we will aim to enroll 4-5 patients of each allowed histology among the first 21 enrolled 
in the ‘other histology’ arm. Although this has its own inherent issues due to limited sample size, 
since we believe that these ‘other histologies’ will behave the same clinically, it will permit us to 
evaluate the different response rates in a limited number of subjects and determine if they differ 
markedly or not. Since this would merely be an exploratory analysis, we will also look at minor 
response as well to help evaluate for hints of efficacy. If the response rates do seem to potentially 
differ markedly by histology, despite our hypothesis that this will not happen, we may consider 
amending the protocol when appropriate to try to restrict enrollment to those histologies with 
stronger evidence of responsiveness. For patients with breast cancer or other chemotherapy-
sensitive tumors (i.e. sarcoma), only responses seen at day 28 and maintained at 4 months will be 
considered a positive response for accrual to the second phase of this study. 
The dose escalation portion of the study may require 6 patients per cohort. For purposes of 
sample size estimation, we will assume that as few as 9 and no more than 25 patients will be 
required to perform the initial safety evaluation. In order to complete the dose escalation phase 
and both phase 2 cohorts, a total of up to 25+82=107 patients may be required (25 + 2 strata with 
a maximum of 41 apiece). Up to 6 patients enrolled at the MTD will count towards the accrual in 


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the appropriate phase 2 strata if they are evaluable for response and if they would be fully 
eligible for enrollment in the phase 2 portion of the trial. Provided that about 4-5 patients per 
month will be able to be enrolled onto this trial, approximately 2 years may be needed to accrual 
the maximum number of required patients. However, as adequate responses to proceed to the 
second stage of accrual may not occur, the trial may end up accruing as few as 9 +42=57 patients 
if no arm demonstrates sufficient patients to accrue a second stage. 
9 
COLLABORATIVE AGREEMENTS  
We have established a Cooperative Research and Development Agreement (CRADAs #02716 
and #03168) with Kite Pharma, Inc., and will be sharing data with them. 
10 HUMAN SUBJECTS PROTECTIONS  
10.1 RATIONALE FOR SUBJECT SELECTION 
The patients to be entered in this protocol have metastatic cancer which is refractory to standard 
therapy, and limited life expectancies. 
Subjects from both genders and 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. Efforts 
will be made to extend accrual to a representative population, but in this preliminary study, a 
balance must be struck between patient safety considerations and limitations on the number of 
individuals exposed to potentially toxic and/or ineffective treatments on the one hand and the 
need to explore gender and ethnic aspects of clinical research on the other hand. If differences in 
outcome that correlate to gender or to ethnic identity are noted, accrual may be expanded or a 
follow-up study may be written to investigate those differences more fully. 
10.2 PARTICIPATION OF CHILDREN 
The use of the nonmyeloablative regimen in this protocol is a major procedure which entails 
serious discomforts and hazards for the patient, such that fatal complications are possible. It is 
therefore only appropriate to carry out this experimental procedure in the context of life 
threatening metastatic cancer. Since the efficacy of this experimental procedure is unknown, it 
does not seem reasonable to expose children to this risk without further evidence of benefit. 
Should results of this study indicate efficacy in treating metastatic cancer, which is not 
responsive to other standard forms of therapy, future research can be conducted in the pediatric 
population to evaluate potential benefit in that patient population. 
10.3 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 10.5), all subjects ≥ age 18 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 for evaluation. For those subjects that become 
incapacitated and do not have pre-determined substitute decision maker, the procedures 


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described in MAS Policy 87-4 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. 
10.4 EVALUATION OF BENEFITS AND RISKS/DISCOMFORTS 
The experimental treatment has a chance to provide clinical benefit though it is not known if it 
will do so. The risks in this treatment are detailed in section 11. The success of this effort cannot 
be predicted at this time.  
10.5 RISK/BENEFIT ANALYSIS 
Because all patients in this protocol have metastatic cancer, which is refractory to standard 
therapy and limited life expectancies, the potential benefit is thought to outweigh the potential 
risks. 
10.6 CONSENT PROCESS AND DOCUMENTATION 
If the patient has a tumor that is found to be HLA-DP4 positive and MAGE-A3 positive by 
immunohistochemistry, the patient is consented on protocol 03-C-0277, Cell Harvest and 
Preparation for Surgery Branch Adoptive Cell Therapy Protocols. If the lymphocytes can be 
generated for infusion and the patient meets the thorough screening for eligibility, the patient, 
with family members or friends at the request of the patient, 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 is reassured that participation on trial is entirely 
voluntary and that he/she can withdraw or decide against treatment at any time without adverse 
consequences. The Principal Investigator, associate investigator, or clinical fellow is responsible 
for obtaining written consent from the patient. 
10.6.1 Informed consent 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. 
Unless the PI is fluent in the prospective subject’s language, an interpreter will be present to 
facilitate the conversation (using either the long translated form or the short form). 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. 


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11 PHARMACEUTICAL INFORMATION  
11.1 INVESTIGATIONAL INVESTIGATION 
11.1.1 Cell Preparation (ANTI-MAGE-A3-DP4 TRANSDUCED PBL) 
The procedure for expanding the human PBL and the certificate of analysis 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 certificate of analysis is included in Appendix 
5. The PBL will be transduced with retroviral supernatant containing the alpha chain and beta 
chain genes of the Anti-MAGE-A3-DP4.  
Note: Penicillin, Streptomycin, and gentamycin will not be used in the manufacture of products 
for patients with documented allergies to these drugs.  
11.1.1.1 Retroviral Vector Containing the Anti-Mage-A3-DP4 Gene 
The retroviral vector supernatant [PG13-MAGE-A3 TCR encoding a T cell receptor directed 
against MAGE-A3-DP4, was prepared and preserved following cGMP conditions in the Surgery 
Branch Vector Production Facility (SBVPF). The retroviral vector, PG13-MAGE-A3 6F9mC 
TCR (B8), consists of 7,382 bps including the 5' LTR from the murine stem cell virus 
(promoter), packaging signal including the splicing donor (SD) and splicing acceptor sites, alpha 
chain and beta chain genes of the anti-MAGE-A3-DP4 TCR. The alpha and beta chains are 
linked by a P2A peptide. For clone selection, the physical titer will be determined by RNA dot 
blot. The physical titer of the clinical vector will be measured in transducing units per mL 
following a titration on human PBL according to sponsor protocols. The supernatant has been 
stored at the SBVPF upon the completion of production at –80 °C. A portion of the supernatant 
will be shipped on dry ice and stored –80 °C at Fisher Bioservices, Rockville, MD. This storage 
facility is equipped with around-the-clock temperature monitoring. Upon request, supernatant 
will be delivered on dry ice to be used in in vitro transduction. 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.  
11.1.2 Interleukin-2 (Aldesleukin, Proleukin, Recombinant Human Interleukin 2) 
How Supplied: Interleukin-2 (aldesleukin) will be provided by the NIH Clinical Pharmacy 
Department from commercial sources.  
Formulation/Reconstitution: Aldesleukin, NSC #373364, is provided as single-use vials 
containing 22 million IU (−1.3 mg) IL-2 as a sterile, white to off-white lyophilized cake plus 50 
mg 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. 
Storage: Intact vials are stored in the refrigerator (2° - 8°C) protected from light. Each vial bears 
an expiration date. 


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Dilution/Stability: Reconstituted aldesleukin should be further diluted with 50 mL of 5% Human 
Serum Albumin (HSA). The HSA should be added to the diluent prior to the addition of RIL-2. 
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° – 30°C. 
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.  
Toxicities: Expected toxicities of aldesleukin are listed in the product label and in Appendix 2 
and Appendix 3. Grade 3 toxicities common to aldesleukin include diarrhea, nausea, vomiting, 
hypotension, skin changes, anorexia, mucositis, dysphagia, or constitutional symptoms and 
laboratory changes as detailed in Appendix 2. Additional grade 3 and 4 toxicities seen with 
aldesleukin are detailed in Appendix 3. 
11.1.3 Fludarabine 
(Please refer to package insert for complete product information)  
Description: Fludarabine phosphate is a synthetic purine nucleoside that differs from physiologic 
nucleosides in that the sugar moiety is arabinose instead of ribose or deoxyribose. Fludarabine is 
a purine antagonist antimetabolite. 
How Supplied: It will be purchased by the NIH Clinical Pharmacy Department from commercial 
sources. Fludarabine is supplied in a 50 mg vial as a fludarabine phosphate powder in the form of 
a white, lyophilized solid cake. 
Stability: Following reconstitution with 2 mL of sterile water for injection to a concentration of 
25 mg/mL, the solution has a pH of 7.7. The fludarabine powder is stable for at least 18 months 
at 2-8°C; when reconstituted, fludarabine is stable for at least 16 days at room temperature. 
Because no preservative is present, reconstituted fludarabine will typically be administered 
within 8 hours. Specialized references should be consulted for specific compatibility 
information. Fludarabine is dephosphorylated in serum, transported intracellularly and converted 
to the nucleotide fludarabine triphosphate; this 2-fluoro-ara-ATP molecule is thought to be 
required for the drug’s cytotoxic effects. Fludarabine inhibits DNA polymerase, ribnucleotide 
reductase, DNA primase, and may interfere with chain elongation, and RNA and protein 
synthesis. 
Storage: Intact vials should be stored refrigerated (2-8°C). 
Administration: Fludarabine is administered as an IV infusion in 100 mL 0.9% sodium chloride, 
USP over 15 to 30 minutes. The doses will be based on body surface area (BSA). If patient is 
obese (BMI > 35) drug dosage will be calculated using practical weight as described in 
Appendix 1. 
Toxicities: At doses of 25 mg/m2/day for 5 days, the primary side effect is myelosuppression; 
however, thrombocytopenia is responsible for most cases of severe and life-threatening 
hematologic toxicity. Serious opportunistic infections have occurred in CLL patients treated with 
fludarabine. Hemolytic anemia has been reported after one or more courses of fludarabine with 
or without a prior history of a positive Coomb’s test; fatal hemolytic anemia has been reported. 
In addition, bone marrow fibrosis has been observed after fludarabine therapy. Other common 
adverse effects include malaise, fever, chills, fatigue, anorexia, nausea and vomiting, and 


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weakness. Irreversible and potentially fatal central nervous system toxicity in the form of 
progressive encephalopathy, blindness, and coma is only rarely observed at the currently 
administered doses of fludarabine. More common neurologic side effects at the current doses of 
fludarabine include weakness, pain, malaise, fatigue, paresthesia, visual or hearing disturbances, 
and sleep disorders. Adverse respiratory effects of fludarabine include cough, dyspnea, allergic 
or idiopathic interstitial pneumonitis. Tumor lysis syndrome has been rarely observed in 
fludarabine treatment of CLL. Treatment on previous adoptive cell therapy protocols in the 
Surgery Branch have caused persistently low (below 200) CD4 counts, and one patient 
developed polyneuropathy manifested by vision blindness, and motor and sensory defects. 
11.1.4 Cyclophosphamide 
(Refer to FDA-approved package insert for complete product information): 
Description: Cyclophosphamide is a nitrogen mustard-derivative alkylating agent. Following 
conversion to active metabolites in the liver, cyclophosphamide functions as an alkyating agent; 
the drug also possesses potent immunosuppressive activity. The serum half-life after IV 
administration ranges from 3-12 hours; the drug and/or its metabolites can be detected in the 
serum for up to 72 hours after administration.  
How Supplied: Cyclophosphamide will be obtained from commercially available sources by the 
NIH Clinical Center Pharmacy Department. 
Stability: Following reconstitution as directed with sterile water for injection, cyclophosphamide 
is stable for 24 hours at room temperature or 6 days when kept at 2-8°C.  
Administration: It will be diluted in 250 mL D5W and infused over one hour. The dose will be 
based on the patient’s body weight. If patient is obese (BMI > 35) drug dosage will be calculated 
using practical weight as described in Appendix 1. 
Toxicities: Hematologic toxicity occurring with cyclophosphamide usually includes leukopenia 
and thrombocytopenia. Anorexia, nausea and vomiting, rash and alopecia occur, especially after 
high-dose cyclophosphamide; diarrhea, hemorrhagic colitis, infertility, and mucosal and oral 
ulceration have been reported. Sterile hemorrhagic cystitis occurs in about 20% of patients; 
severity can range from microscopic hematuria to extensive cystitis with bladder fibrosis. 
Although the incidence of hemorrhagic cystitis associated with cyclophosphamide appears to be 
lower than that associated with ifosfamide, mesna (sodium 2-mercaptoethanesulfonate) has been 
used prophylactically as a uroprotective agent in patients receiving cyclophosphamide. 
Prophylactic mesna is not effective in preventing hemorrhagic cystitis in all patients. Patients 
who receive high dose cyclophosphamide may develop interstitial pulmonary fibrosis, which can 
be fatal. Hyperuricemia due to rapid cellular destruction may occur, particularly in patients with 
hematologic malignancy. Hyperuricemia may be minimized by adequate hydration, 
alkalinization of the urine, and/or administration of allopurinol. If allopurinol is administered, 
patients should be watched closely for cyclophosphamide toxicity (due to allopurinol induction 
of hepatic microsomal enzymes). At high doses, cyclophosphamide can result in a syndrome of 
inappropriate antidiuretic hormone secretion; hyponatremia with progressive weight gain without 
edema occurs. At high doses, cyclophosphamide can result in cardiotoxicity. Deaths have 
occurred from diffuse hemorrhagic myocardial necrosis and from a syndrome of acute 
myopericarditis; in such cases, congestive heart failure may occur within a few days of the first 
dose. Other consequences of cyclophosphamide cardiotoxicity include arrhythmias, potentially 


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irreversible cardiomyopathy, and pericarditis. Other reported adverse effects of 
cyclophosphamide include headache, dizziness, and myxedema; faintness, facial flushing, and 
diaphoresis have occurred following IV administration. Mesna (sodium 2-
mercaptoethanesulphonate; given by IV injection) is a synthetic sulfhydryl compound that can 
chemically interact with urotoxic metabolites of cyclophosphamide (acrolein and 4-
hydroxycyclophosphamide) to decrease the incidence and severity of hemorrhagic cystitis.  
11.2 SUPPORTIVE MEDICATIONS 
11.2.1 Mesna  
(Sodium 2-mercaptoethanesulfonate, Mesnum, Mesnex, NSC-113891)  
(Please refer to the FDA-approved package insert for complete product information) 
Description: Mesna will be obtained commercially by the Clinical Center Pharmacy Department 
and is supplied as a 100 mg/mL solution.  
Storage: Intact ampoules are stored at room temperature.  
Stability: Diluted solutions (1 to 20 mg/mL) are physically and chemically stable for at least 24 
hours under refrigeration. Mesna is chemically stable at room temperature for 48-72 hours in 
D5W, 48-72 hour in D5W/0.45% NaCl, or 24 hours in 0.9% NaCl. 
Administration: Dilute to concentrations less than or equal to 20 mg mesna/mL fluid in D5W or 
0.9% NaCl and to be administered intravenously as a continuous infusion. If patient is obese 
(BMI > 35) drug dosage will be calculated using practical weight as described in Appendix 1. 
Toxicities include nausea, vomiting and diarrhea. 
11.2.2 Filgrastim 
(Granulocyte Colony-Stimulating Factor, G-CSF, Filgrastim, Neupogen) 
Filgrastim will be obtained commercially by the Clinical Center Pharmacy Department and is 
supplied in 300 ug/mL and 480 ug/1.6 mL vials. G-CSF should be refrigerated and not allowed 
to freeze. The product bears the expiration date. The product should not be shaken. It is generally 
stable for at least 10 months when refrigerated. The appropriate dose is drawn up into a syringe. 
G-CSF will be given as a daily subcutaneous injection. The side effects of G-CSF are skin rash, 
myalgia and bone pain, an increase of preexisting inflammatory conditions, enlarged spleen with 
occasional associated low platelet counts, alopecia (with prolonged use) elevated blood 
chemistry levels. 
11.2.3 Trimethoprim and Sulfamethoxazole Double Strength (TMP / SMX DS): 
TMP/SMX DS will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used for the prevention of PCP pneumonia. The oral dose is 1 tablet PO daily 
three times a week (MUST be on non-consecutive days) beginning on day 0 or within one week 
of anticipated lymphopenia and continuing for at least 6 months and until the CD4 count is 
greater than 200 on 2 consecutive lab studies. Like other sulfa drugs, TMP/SMX DS can cause 
allergies, fever, photosensitivity, nausea, and vomiting. Allergies typically develop as a 
widespread itchy red rash with fever eight to fourteen days after beginning the standard dose. 
Neutropenia, a reduction in the number of neutrophils, can also occur. Should allergies develop, 
the following medications may be used in place of TMP/SMX DS. 


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11.2.3.1 Dapsone 
Dapsone will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used for the prevention of Pneumocystis pneumonia. The dose is 100mg by 
mouth daily, starting on day 0 (± 7 days) and continuing at least 6 months and until the CD4+ 
count is > 200 on two consecutive lab studies. It is supplied as 25mg and 100mg tablets. 
Dapsone contains a sulfa group, although the cross reactivity in patients with sulfa allergies is 
quite low. Dapsone may be considered in patients with mild to moderate sulfa allergies. Dapsone 
should be avoided in patients with severe (i.e., a history of anaphylaxis or other equally serious 
reaction) reactions to sulfa drugs. Additionally, dapsone has been reported to cause hemolytic 
anemia is patients with G6PD deficiency. It is recommended that patients be tested for G6PD 
deficiency prior to the initiation of dapsone therapy. Dapsone is generally well tolerated, but may 
cause a number of hematologic adverse reactions, including increased reticulocyte counts, 
hemolysis, decreased hemoglobin, methemoglobinemia, agranulocytosis, anemia, and 
leukopenia. Other rare but serious adverse reactions include bullous exfoliative dermatitis, 
Stevens-Johnson syndrome, toxic epidermal necrolysis, pancreatitis, interstitial pneumonitis, and 
pulmonary eosinophilia. For more detailed information about adverse reactions, consult the 
package insert. 
11.2.3.2 Atovaquone 
Atovaquone will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used for the prevention of Pneumocystis pneumonia in patients who cannot 
tolerate or are allergic to sulfamethoxazole/trimethoprim, dapsone, or pentamidine. Atovaquone 
may be given as a single daily dose of 1500mg orally or the dose may be split into 750mg given 
orally twice daily. Atovaquone will be started on day 0 (± 7 days), and will continue for at least 6 
months and until the CD4+ count is > 200 on two consecutive lab studies. Atovaquone is 
supplied as an oral suspension containing 150mg/mL. Common adverse reactions to atovaquone 
include: headache, rash, diarrhea, nausea, vomiting, abdominal pain, cough, and fever. Rare but 
serious adverse reactions include acute renal failure, hepatitis and hepatic failure, angioedema, 
pancreatitis, and Stevens-Johnson syndrome. For more detailed information about adverse 
reactions, consult the package insert. 
11.2.3.3 Aerosolized Pentamidine 
Patients with sulfa allergies will receive aerosolized Pentamidine 300 mg per nebulizer with one 
week prior to admission and continued monthly until the CD4 count is above 200 on two 
consecutive follow up lab studies and for at least 6 months post chemotherapy. Pentamidine 
Isethionate will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used to prevent the occurrence of PCP infections. It is supplied in 300 mg vials 
of lyophilized powder and will be administered via nebulizer. Toxicities reported with the use of 
Pentamidine include metallic taste, coughing, bronchospasm in heavy smokers and asthmatics; 
increased incidence of spontaneous pneumothorax in patients with previous PCP infection or 
pneumatoceles, or hypoglycemia.  
11.2.4 Herpes and Varicella Zoster Virus Prophylaxis 
11.2.4.1 Valacyclovir (Valtrex) 
Valacyclovir will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used orally to prevent the occurrence of herpes virus infections in patients with 
positive HSV serology. It is supplied in 500 mg tablets. Valcyclovir will be started at a dose of 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
50
500 mg orally daily if the patient is able to tolerate oral intake. See package insert for dosing 
adjustments in patients with renal impairment. Common side effects include headache, upset 
stomach, nausea, vomiting, diarrhea or constipation. Rare serious side effects include hemolytic 
uremic syndrome and thrombotic thrombocytopenic purpura. 
11.2.4.2 Acyclovir 
Acyclovir will be obtained by the Clinical Center Pharmacy Department from commercial  
sources. It will be used to prevent the occurrence of herpes virus infections in patients who 
cannot take oral medications. It is supplied as powder for injection in 500 mg/vials. Reconstitute 
in 10 mL of sterile water for injection to a concentration of 50 mg/mL. Reconstituted solutions 
should be used within 12 hours. IV solutions should be diluted to a concentration of 7mg/mL or 
less and infused over 1 hour to avoid renal damage. Reversible renal insufficiency has been 
reported with IV but not oral acyclovir. Neurologic toxicity including delirium, tremors, coma, 
acute psychiatric disturbances, and abnormal EEGs have been reported with higher doses of 
acyclovir. Should this occur, a dosage adjustment will be made or the drug will be discontinued. 
Stomach upset, headache or nausea, rash or hives; peripheral edema; pain, elevated liver function 
tests; and leukopenia, diarrhea, lymphadenopathy, myalgias, visual abnormalities and elevated 
creatinine have been reported. Hair loss from prolonged use has been reported. Acyclovir will 
not be used concomitantly with other nucleoside analogs which interfere with DNA synthesis, 
e.g. ganciclovir. In renal disease, the dose is adjusted as per product labeling. 
11.2.5 Fluconazole 
Fluconazole will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used to prophylax against fungal infections. It is available in 200 mg tablets. It 
can cause headache, nausea, vomiting, diarrhea or abdominal pain, and liver damage which may 
be irreversible. It can cause rashes and itching, which in rare cases has caused Stevens Johnson 
Syndrome. It has several significant drug interactions. The package insert should be consulted 
prior to prescribing. For IV administration in patients who cannot tolerate the oral preparation, 
Fluconazole comes in 2 mg/mL solution for injection, and prepared according to Clinical Center 
Pharmacy standard procedures. It should be administered at a maximum IV rate of 200 mg/hr. 
11.2.6 Ondansetron hydrochloride 
Ondansetron hydrochloride will be obtained by the Clinical Center Pharmacy Department from 
commercial sources. It will be used to control nausea and vomiting during the chemotherapy 
preparative regimen. It can cause headache, dizziness, myalgias, drowsiness, malaise, and 
weakness. Less common side effects include chest pain, hypotension, pruritis, constipation and 
urinary retention. Consult the package insert for specific dosing instructions. 
11.2.7 Furosemide 
Furosemide will be obtained by the Clinical Center Pharmacy Department from commercial 
sources. It will be used to enhance urine output during the chemotherapy preparative regimen 
with cyclophosphamide. Adverse effects include dizziness, vertigo, paresthesias, weakness, 
orthostatic hypotension, photosensitivity, rash and pruritis. Consult the package insert for a 
complete list of all side effects. 
 
 


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55
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13 FIGURES, TABLES & APPENDICES 
Figure 1  
 


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Figure 1. Comparison of reactivity of T cells transduced with three TCR that recognize the 
MAGE-A3:245-258 HLA class II restricted epitope (a) PBMC from three melanoma patients 
were stimulated with OKT3 and transduced on day 2, and evaluated six days later for expression 
of the transduced TCRs using the appropriate anti-TCR Vβ antibodies. Un-transduced cells 
incubated with the anti-Vβ22 antibody demonstrated relatively low levels of endogenous cell 
surface expression of this TCR. (b) CD4+ T cells from the three patient PBMC that had been 
transduced with the 6F9, R12C9 and 12C7 TCRs were isolated by using a CD4 T lymphocyte 
enrichment kit (Life Technologies, Grand Island, NY). The isolated T cells (105) were cultured 
with either HLA-DP*0401+ 293-CITTA cells (105) that were transiently transfected with plasmid 
constructed encoding individual MAGE family members, 293-CIITA cells that were pulsed with 
the 10 µg/mL of the MAGE-A3:245-258 peptide. In addition, transduced CD4+ T cells were 
cultured with 105 1359 mel-CIITA cells, which express MAGE-A3 as well as HLA-DP*0401, or 
624 mel-CIITA cells, which express MAGE-A3 but do not express HLA-DP*0401 or 0402. 
Following an overnight co-culture, the levels of soluble IFN-γ were evaluated by ELISA. (c) 
CD4+ T cells transduced with the three MAGE-A3 TCRs (105) were incubated with 293-CIITA 
cells (105) that were pulsed with the indicated concentrations of the MAGE-A3:245-258 peptide 
for two hours at 370C. Following the incubation, the target cells were washed twice and co-
cultured overnight with CD4+ T cells from PBMC 1, and the release of soluble IFN-γ measured 
by ELISA. 
 
 


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Figure 2 
 


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Figure 2. Comparison of responses of CD4+ and CD8+ T cells transduced with the 6F9 TCR or 
the 6F9 TCR containing murine constant region sequences (6F9mc). (a) Evaluation of responses 
of CD4+ and CD8+ T cells isolated from two patient PBMC against MAGE-A3+ and MAGE-A3- 
tumor cells that express HLA-DP*0401, 0402, or lack expression of either HLA-DP allele. 
Targets include 8 melanoma cell lines (2630 mel-CIITA, 397 mel-CIITA,2984-CIITA, 526 mel-
CIITA, 1359 mel-CIITA, SK37 mel-CIITA, 3071 mel-CIITA, and 624 mel-CIITA), one non-
small cell lung carcinoma (H1299 NSCLC-CIITA), and one renal cancer cell line (1764 RCC-
CIITA). (b) Evaluation of responses of CD4+ T cells that were transduced with either the 6F9 or 
the 6F9mc TCR against tumor targets. (c) Evaluation of responses of CD8+ T cells that were 
transduced with either the 6F9 or the 6F9mc TCR against tumor targets. Co-cultures were carried 
out as described in Figure 1. 
 
 


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Figure 3 
 
 
 
 


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Figure 3. Flow cytometric analysis of T cells for patient infusion. Peripheral blood lymphocytes 
were CD8+ T cells depleted on day 0 and transduced with MAGE-A3 DP4 TCR on day 2 and 3. 
Cells were subjected to flow cytometric analysis on day 9 (A). Cells on day 13 after rapid 
expansion were also subjected to flow cytometric analysis (B). Unseparated, un-transduced 
lymphocytes functioned as the negative control. Representative data from one of the three different 
donors are shown. 
 
 
 
 
 


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Figure 4 
 
 
  A  
 
 
 
 
 
 
 
  B 
 
 
 
 
 
 
 
 


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Figure 4. Recognition of MAGE-A3+, DP0401+ tumor cell line and MAGE-A3 DP4 peptide-
pulsed cells. On day 10 after CD8+ T cell depletion (A) or day 13 after rapid expansion (B), 
MAGE-A3 DP4 TCR-transduced CD4+ T cells were co-cultured for 16 hr with tumor target cell 
lines (2630 CIITA: MAGE-A3+, DP0401+; 624 CIITA: MAGE-A3+, DP0401-; 1764 CIITA: 
MAGE-A3-, DP0401+) or MAGE-A3 DP4 peptide-pulsed 293 CIITA cells. The secretion of IFN-
gamma was determined by ELISA. Representative data from one of the three different donors are 
shown. 
 
 


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Table 1 
Table 1. Evaluation of response to peptides related to MAGE-A3:243-258  
a
b
Amino acids 243-258
MAGE-A3
KKLLTQHFVQENYLEY
MAGE-A6
KKLLTQYFVQENYLEY
MAGE-A1
KKLLTQDLVQEKYLEY
MAGE-A12 KKLLTQDLVQENYLEY
*CD4+ T cells isolated from two patient PBMC by negative selection were transduced with  either 
the  6F9 TCR, the 6F9mc TCR, or were un-transduced  and assayed 10 days following OKT3 
stimulation for their response to 293-CIITA cells that were pulsed with 10 µg/ml of the indicated 
peptides, as described in Figure 1.
PBMC-1* transduced with:
PBMC-2  transduced with:
6F9 
6F9mc 
None
6F9
6F9mc
None
Predicted
Gene (position) Amino Acid Sequence
IFN-γ (pg/ml)
affinity(nM)
MAGE-A3:243-258 KKLLTQHFVQENYLEY
10,220
15,210
33
10,350
17,520
45
3
MAGE-A3:243-256 KKLLTQHFVQENYL  
1,018
1,815
72
1,670
2,490
78
323
MAGE-A3:243-255 KKLLTQHFVQENY   
76
137
29
111
117
71
378
MAGE-A3:243-254 KKLLTQHFVQEN
28
0
67
30
39
78
466
MAGE-A3:243-253 KKLLTQHFVQE
0
40
38
30
45
90
2444
MAGE-A3:245-258
LLTQHFVQENYLEY
9,290
14,970
84
8,920
17,820
74
3
MAGE-A3:246-258
LTQHFVQENYLEY
7,140
12,700
56
9,200
16,170
76
3
MAGE-A3:247-258
TQHFVQENYLEY
6,710
10,600
30
6,810
13,280
41
3
MAGE-A3:248-258
QHFVQENYLEY
6,220
9,000
52
7,400
8,700
56
4
MAGE-A3:249-258
HFVQENYLEY
669
1,643
57
922
2,034
66
5
MAGE-A6:248-258
QYFVQENYLEY
6,440
11,800
54
13,200
8,370
127
3
MAGE-A2/A12:248-258
QDLVQENYLEY
33
66
49
37
56
65
59
MAGE-A4/A9:249-259
QDWVQENYLEY
0
23
32
22
26
62
92
MAGE-A8:251-261
QEWVQENYLEY
43
58
79
39
41
55
87
MAGE-A1/B4:241-251
QDLVQEKYLEY
129
126
55
108
84
53
16
MAGE-B2:250-260
KDLVQEKYLEY
0
0
43
7
20
38
16
MAGE-B10:250-260
KDLVKENYLEY
22
18
69
28
34
66
105
MAGE-B16:252-262
KDFVKEKYLEY
0
27
16
11
28
42
3
MAGE-C1:113-123
KVWVQEHYLEY
9
0
30
25
27
30
35
MAGE-D4:300-315 RKLITDDFVKQKYLEY
193
234
81
194
268
80
6
MAGE-D2:413-428 KKLITDEFVKQKYLDY
82
43
56
226
223
71
8
MAGE-L2:582-597 KKLITEVFVRQKYLEY
45
56
58
78
107
114
6
MAGE-G1:220-235 KKLITEDFVRQRYLEY
0
29
68
25
33
62
3
Necdin:237-247
EEFVQMNYLKY
0
22
59
21
32
83
13
No peptide
0
5
58
15
25
59
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
65
Appendix 1 
Modification of Dose Calculations* in patients whose BMI is greater than 35 
Unless otherwise specified in this protocol, actual body weight is used for dose 
calculations of treatment agents. In patients who are determined to be obese (BMI > 
35), the practical weight (see 3 below) will be used. 
 
 
1. BMI Determination: 
 
BMI = weight (kg) / [height (m)]2 
 
 
2. Calculation of ideal body weight 
 
Male = 50 kg + 2.3 (number of inches over 60 inches) 
Example: ideal body weight of 5’10’’ male 
50 + 2.3 (10) = 73 kg 
 
Female = 45.5 kg + 2.3 (number of inches over 60 inches) 
Example: ideal body weight of 5’3’’ female 
45.5 + 2.3 (3) = 57 kg 
 
3. Calculation of “practical weight” 
 
Calculate the average of the actual and the ideal body weights. This is the 
practical weight to be used in calculating the doses of chemotherapy and 
associated agents designated in the protocol. 
 
 
 
 
 
 
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
66
 
Appendix 2 
Adverse Events Occurring In ≥10% 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 tachycardia  12  
 
Somnolence   
 
22 
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. 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
67
c Respiratory disorder: ARDS, CXR infiltrates, unspecified pulmonary changes. 
1Source: Proleukin® Prescribing Information – June 2007 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
68
Appendix 3 
Expected IL-2 Toxicities and their Management 
Expected toxicity 
Expected 
grade 
Supportive Measures 
Stop Cycle* 
 
Stop 
Treatment ** 
Chills 
3 
IV Meperidine 25-50 
mg, IV q1h, prn, 
No 
No 
Fever 
3 
Acetaminophen 650 
mg, po, q4h; 
Indomethicin 50-75 
mg, po, q8h 
No 
No 
Pruritis 
3 
Hydroxyzine HCL 
10-20 mg po q6h, 
prn; 
Diphenhydramine 
HCL25-50 mg, po, 
q4h, prn 
No 
No 
Nausea/ Vomiting/ 
Anorexia 
3 
Ondansetron 10 mg, 
IV, q8h, prn; 
Granisetron 0.01 
mg/kg IV daily prn; 
Droperidol 1 mg, IV 
q4-6h, prn; 
Prochlorperazine 25 
mg pr, prn or 10 mg 
IV q6h prn 
No 
No 
Diarrhea 
3 
Loperamide 2mg, po, 
q3h, prn; 
Diphenoxylate HCl 
2.5 mg and atropine 
sulfate 25 mcg, po, 
q3h, prn; codeine 
sulfate 30-60 mg, po, 
q4h, prn 
If uncontrolled after 24 
hours despite all 
supportive measures 
No 
Malaise 
3 or 4 
Bedrest 
If other toxicities occur 
simultaneously 
No 
Hyperbilirubinemia 
3 or 4 
Observation 
If other toxicities occur 
simultaneously 
No 
Anemia 
3 or 4 
Transfusion with 
PRBCs 
If uncontrolled despite 
all supportive measures 
No 
Thrombocytopenia 
3 or 4 
Transfusion with 
platelets 
If uncontrolled despite 
all supportive measures 
No 
Edema/Weight gain 3 
Diuretics prn 
No 
No 
Hypotension 
3 
Fluid resuscitation 
Vasopressor support 
If uncontrolled despite 
all supportive measures 
No 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
69
Dyspnea 
3 or 4 
Oxygen or 
ventilatory support 
If requires ventilatory 
support 
No 
Oliguria 
3 or 4 
Fluid boluses or 
dopamine at renal 
doses 
If uncontrolled despite 
all supportive measures 
No 
Increased creatinine 3 or 4 
Observation 
Yes (grade 4) 
No 
Renal failure 
3 or 4 
Dialysis 
Yes 
Yes 
Pleural effusion 
3 
Thoracentesis 
If uncontrolled despite 
all supportive measures 
No 
Bowel perforation 
3 
Surgical intervention 
Yes 
Yes 
Confusion 
3 
Observation 
Yes 
No 
Somnolence 
3 or 4 
Intubation for airway 
protection 
Yes 
Yes 
Arrhythmia 
3 
Correction of fluid 
and electrolyte 
imbalances; chemical 
conversion or 
electrical conversion 
therapy 
If uncontrolled despite 
all supportive measures 
No 
Elevated Troponin 
levels 
3 or 4 
Observation 
Yes 
If changes in 
LV function 
have not 
improved to 
baseline by 
next dose 
Myocardial 
Infarction 
4 
Supportive care 
Yes 
Yes 
Elevated 
transaminases 
3 or 4 
Observation 
For grade 4 without 
liver metastases 
If changes 
have not 
improved to 
baseline by 
next dose 
Hyperbilirubinemia 
3 or 4 
Observation 
For grade 4 without 
liver metastases 
If changes 
have not 
improved to 
baseline by 
next dose 
Electrolyte 
imbalances 
3 or 4 
Electrolyte 
replacement 
If uncontrolled despite 
all supportive measures 
No 
Neutropenia 
4 
Observation 
No 
No 
*Unless the toxicity is not reversed within 12 hours 
** Unless the toxicity is not reversed to grade 2 or less by next treatment. 
 
 
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
70
Appendix 4 
 
Interleukin-2 toxicities observed in patients treated at the NIH Clinical Center 
 
 
 
 
 
 
 


Abbreviated Title: Anti-MAGE-A3-DP4 TCR PBL 
Version Date: June 22, 2018 
 
71
Appendix 5 
 
Certificate of Analysis 
Infused T cells transduced with MAGE-A3 DP0401/0402 TCR genes. 
Date of preparation of final product: 
Patient: 
Tests performed on final product: 
 
Test 
 
Method 
Limits 
Result 
Tests 
Performed 
by 
Initials/ 
Date 
Cell viability1 
trypan blue exclusion 
>70% 
 
 
 
Total viable cell 
number1 
visual microscopic count >1 x107 
 
 
 
Tumor reactivity2 
γ-IFN release vs. peptide 
pulsed 293-CIITA 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 
negative 
 
 
 
Endotoxin  
limulus assay1 
≤ 5 E.U./kg 
 
 
 
RCR 
S+L- Assay4 
RCR-PCR6 
 
negative 
 
 
 
1 Performed on sample of the final product immediately prior to infusion. Results are available at the time of infusion. 
2 Performed 2-10 post transduction. Results are available at the time of infusion.  
3 Performed 2-4 days prior to infusion. Results are available at the time of infusion but may not be definitive. 
4 Sample collected from the final product prior to infusion. Results will not be available before cells are infused into the patient. 
5 Performed 2-10 days prior to infusion. Results are available at the time of infusion. 
6 Performed on sample approximately 1-4 days prior to infusion. Results are available at the time of infusion.  
 
Prepared by: 
_______________________________________ Date: ____________________ 
 
 
QC sign-off: 
_______________________________________ Date: ____________________ 
Qualified Clinical or Laboratory Supervisor 
 
 

ICF_001.pdf: 
MEDICAL RECORD 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
• 
Adult Patient or 
• Parent, for Minor Patient 
INSTITUTE: 
National Cancer Institute 
STUDY NUMBER: 
14-C-0052 
PRINCIPAL INVESTIGATOR: 
Steven A. Rosenberg, M.D., Ph.D
STUDY TITLE:  
 
Phase I/II Study of the Treatment of Metastatic Cancer that Expresses MAGE-A3 Using 
Lymphodepleting Conditioning Followed by Infusion of HLA-DP0401/0402 Restricted 
Anti-MAGE-A3 TCR-Gene Engineered Lymphocytes and Aldesleukin 
 
Continuing Review Approved by the IRB on 06/24/19 
Amendment Approved by the IRB on 06/27/18 (K) 
 
                        Date posted to web: 07/02/19 
Standard 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
• 
Adult Patient or 
• Parent, for Minor Patient 
NIH-2514-1 (07-09) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent (1) 
 
 Revision Copy
INTRODUCTION 
We invite you to take part in a research study at the National Institutes of Health (NIH). 
First, we want you to know that: 
Taking part in NIH research is entirely voluntary.  
 
You may choose not to take part, or you may withdraw from the study at any time.  In 
either case, you will not lose any benefits to which you are otherwise entitled.  However, 
to receive care at the NIH, you must be taking part in a study or be under evaluation for 
study participation. 
You may receive no benefit from taking part.  The research may give us knowledge that 
may help people in the future. 
Second, some people have personal, religious or ethical beliefs that may limit the kinds of medical 
or research treatments they would want to receive (such as blood transfusions).  If you have such 
beliefs, please discuss them with your NIH doctors or research team before you agree to the study. 
Now we will describe this research study.  Before you decide to take part, please take as much 
time as you need to ask any questions and discuss this study with anyone at NIH, or with family, 
friends or your personal physician or other health professional.   
 
Why is this study being done?  
We have developed an experimental therapy for treating patients with cancer that involves taking 
white blood cells from the patient, selecting a specific type of white blood cell to grow in the 
laboratory in large numbers, genetically modifying these specific cells with a type of virus 
(retrovirus) to attack only the tumor cells, and then giving the cells back to the patient.  This type 
of therapy is called gene transfer.   In this protocol we are modifying the patient’s white blood 
cells with a retrovirus that has the gene for anti-MAGE-A3-DP4 incorporated in the retrovirus.  So 
far, MAGE-A3-DP4 has been found only on tumor cells.    The purpose of this study is to see if 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
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NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 2 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
these particular tumor fighting cells can cause tumors to shrink and to be certain the treatment is 
safe.  However, it is possible that these cells will not have this effect.    The first few patients 
enrolled will participate in the Phase I portion of the study, called the “dose escalation” phase.  The 
purpose of dose escalation is to determine the most effective yet safe dose of anti-MAGE-A3-DP4 
cells.  There will be 9 dose levels of anti-MAGE-A3-DP4 cells. The first patients enrolled get the 
smallest dose and the dose is increased when a level has been determined to be safe.  If the first 
dose has too many side effects, patients enrolled will receive smaller doses.  Discuss with your 
doctor which dose of anti-MAGE-A3-DP4 cells you will be receiving. This is the first time these 
cells will be given to humans.  Since this is a phase I/II trial, it is unlikely that you will receive any 
benefit. 
 
Why are you being asked to take part in this study? 
You are being asked to participate in this study because you have been diagnosed with cancer that 
has the MAGE-A3-DP4 molecule on the surface of the tumors. 
 
How many people will take part in this study?  
Up to 107 patients will be enrolled in this study.  
 
Description of Research Study 
Stages 1 is performed under the companion protocol, 99-C-0128 (Evaluation for NCI Surgery 
Branch Clinical Research Protocols), to which you have already enrolled. 
Stages 2 is performed under the companion protocol, 03-C-0277 (Cell Harvest and Preparation for 
Surgery Branch Adoptive Cell Therapy Protocols), to which you have already enrolled. 
Stages 3-6 are performed under this protocol. 
This study has the following 5 stages: 
Stage 
Timeframe 
Location 
Comments & Instructions 
1. Work up 
1-2 weeks 
Inpatient or 
out patient   
Scans, x-rays, labs and other 
tests as needed 
2. Chemotherapy 
(day -7 to -3) 
5 days 
Inpatient 
Receive IV chemotherapy to 
prepare your immune system for 
the cells 
3. Cell 
Manufacturing 
4-6 weeks 
Inpatient or 
Outpatient 
Leukapheresis 
4. Cells and 
aldesleukin 
1-5 days 
Inpatient and 
possibly ICU 
Receive the anti-MAGE-A3-
DP4 cells IV and then high dose 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 3 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
(Day 0-4) 
aldesleukin about every 8 hours 
for up to 15 doses 
5. Recovery 
1-3 weeks 
Inpatient unit 
Recover from the effects of 
treatment 
6. Follow -up 
Ongoing 
until disease 
progression 
Outpatient 
Return to clinic for physical 
exam, review of side effects, 
labs, scans every 1-3 months for 
the first year. 
 
 
 
What will happen if you take part in this research study?   
Before you begin the study 
The following procedures are conducted under 99-C-0128 or 03-C-0277. 
Cell harvest and growth 
You underwent a process called apheresis, while enrolled on our companion protocol 03-C-0277 
(Cell harvest and Preparation for Surgery Branch Adoptive Cell Therapy Protocols).  This process 
obtained blood cells from you.  Some of these cells will be grown in the lab and genetically 
modified to recognize a protein on your tumor cells. If your cells do not grow, unfortunately, you 
will not be able to receive the cell infusion.  If that happens, we will look at alternative 
experimental treatments at the NIH Clinical Center or refer you to the care of your referring 
physician.  We usually know after about 4 weeks whether the cells will grow well enough to be 
used as an experimental treatment on this protocol.  At the time we determine that your cells are 
not growing, we will inform you and discuss your options with you. Several medications are used 
during the preparation of your cell product, be sure to tell your doctor if you are allergic to any 
antibiotics. 
 
Work up 
Prior to receiving the experimental treatment, you will undergo many tests. These include imaging 
procedures, heart and lung function tests, eye exams, and laboratory tests.  If you have received 
ipilimumab (also called Yervoy, MDX-010) or any antibody therapy that could affect an anti-
cancer immune response and have experienced any GI toxicities, you will have a colonoscopy and 
biopsies to make sure your colon is normal since these drugs may have caused damage to your 
colon that may worsen with aldesleukin treatment.  If you are a woman, you will undergo a 
pregnancy test.    You may be admitted to the hospital for these tests. However, you will be allowed 
to leave on pass on the days that you are not having tests performed. 
 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
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NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 4 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
Catheter insertion 
Prior to beginning the experimental treatment, you will have an intravenous (IV) catheter placed 
in your upper chest. The area will be numbed with an anesthetic before the catheter is put in.  
Although rare, putting these catheters in can sometimes cause collapse of a lung or cause bleeding.  
Lung collapse is treated by putting a tube into your chest for a few days to allow your lung to 
expand.  Pressure is placed on any area that might bleed.  Other IVs may be needed in one or both 
of your arms if we need to give you extra fluids, medicines, or nutrition. 
 
Leukapheresis 
Leukapheresis is a procedure that allows us to remove certain types of blood cells from you and 
return the rest of your blood.  It is a very common procedure that is done routinely here at the NIH 
with very few risks.  During leukapheresis, blood is removed from you through a needle in your 
arm, circulated through a machine that divides whole blood into red cells, plasma (the serum part), 
and lymphocytes (or white cells), and then the plasma and red cells are returned to you through a 
second needle in your other arm.  The white blood cells may be used to help grow the cells and 
after the treatment, you will have leukapheresis so that we can test your cells. In addition to the 
leukapheresis you will undergo as part of your work up, we will also ask you to undergo one 
additional leukapheresis procedure between 4 and 6 weeks after your cell regimen to see the impact 
of this therapy on the immune system and see if cells we gave you are still active. 
 
During the Study 
The following procedures will be performed under this protocol. 
Chemotherapy Regimen (Day -7 through Day -3) 
After we have grown the anti-MAGE-A3-DP4 cells to large numbers in the laboratory, you will 
be admitted to the hospital to begin your experimental treatment. You will be given two 
chemotherapy medicines, cyclophosphamide and fludarabine, to suppress your immune system so 
the anti-MAGE-A3-DP4 cells can work without any interference from the cells in your immune 
system. (These medicines will not treat your cancer; although they may cause your tumor to shrink 
for a short period of time.)  Animal experiments have indicated that this can make the cells more 
effective in fighting cancer tumors, but it is not known whether this is true in humans.  The 
cyclophosphamide will be given into your catheter over 1 hour for two days (Day -7 and Day -6) 
and the fludarabine will be given into your catheter for 30 minutes every day for the next five days 
(Day -7 through Day -3).  The side effects of these medicines are described on the following pages. 
After you have completed the chemotherapy regimen you will receive the cell infusion  
 
Cell Infusion and IL-2 Regimen (Day 0 through Day 5)  
One to four days after the last dose of chemotherapy, you will be given the anti-MAGE-A3-DP4 
cells.  The anti-MAGE-A3-DP4 cells will be given in your intravenous catheter over 30 minutes.  
Within 24 hours after the anti-MAGE-A3-DP4 cell infusion, you will be given high dose 


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STUDY NUMBER:  14-C-0052 
CONTINUATION: page 5 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
aldesleukin through your intravenous catheter. Aldesleukin is approved by the FDA for treatment 
of metastatic melanoma and metastatic renal cell cancer.  The purpose of giving the aldesleukin 
with this therapy is to keep the cells we give you active for as long as possible so they will fight 
your tumor.  The aldesleukin will be given as a 15-minute infusion about every 8 hours for up to 
five days after the cell infusion (maximum number of doses is 15).  Doses may be skipped or 
delayed depending on how well you tolerate the doses.  The risks of the cells and aldesleukin are 
described on the following pages. 
 
The day after you receive the anti-MAGE-A3-DP4 cells, we will give you G-CSF (filgrastim) as 
a shot or injection under the skin. This will continue until your white blood cell counts begin to 
return to normal.  We will watch you closely during this entire time for any side effects of this 
experimental regimen.  We will discuss the side effects below and we will include in your care all 
the medicines and treatments to prevent as many of these side effects as we can and to make you 
as comfortable as we can.   
 
When you are finished taking the drugs (treatment) 
 
Recovery 
After your last dose of aldesleukin, you will recover in the hospital until you are well enough to 
go home.  This usually takes 7 to 21 days; however, you may need to stay in the hospital for longer 
than this. We will continue to give you support medications, do laboratory tests, and watch you 
closely for any side effects until we feel your condition is stable.  
 
In addition to the laboratory tests to monitor your condition, we will remove approximately 9 
teaspoons of blood three times per week to study the effects of this regimen on your immune 
system.  The maximum amount of blood for research is approximately 2.3 cups in 8 weeks. 
 
 
Follow up and Evaluation of Experimental Regimen  
You will need to continue to take Bactrim, an antibiotic, for at least 6 months following your 
treatment. We will ask you to return to the NIH Clinical Center frequently after you are discharged 
approximately 6 and 12 weeks following treatment and then if you are responding to the treatment, 
every 3 months x3, then every 6 months x2 years and then as determined by your physician.  The 
follow up visits will probably take 1-2 days.  At each visit you will have lab tests, imaging studies 
and a physical examination. At some of your follow up visits, you may undergo leukapheresis or 
have about 8 tubes of blood drawn (4 tablespoons) so that we can see the effect this therapy has 
had on your immune system and if the cells we gave you are still alive. If you are unwilling or 
unable to travel to the NIH Clinical Center we will contact you by phone or e-mail and we may 
ask you to send us lab, imaging, and physical exam reports. If your tumor appears to be growing, 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
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NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 6 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
we will look for other investigational therapies you may be eligible for, or refer you back to the 
care of your local physician. 
 
Retreatment 
If your tumor shrinks or disappears following the initial treatment and then stops shrinking or 
recurs, you may receive additional treatments if you tolerated the treatment well and if all the side 
effects have resolved.    The second treatment will not begin prior to 6- 8 weeks after your last 
dose of IL-2.   
 
Birth Control 
If you are a woman who is breast feeding or pregnant, you may not take part in the study because 
we don’t know how this medicine would affect your baby or your unborn child. If you are a woman 
who can become pregnant, or are the partner of a woman who can become pregnant, you will need 
to practice an effective form of birth control before starting study treatment, during study 
treatment, and for four months after your blood tests show no evidence of the cells. Male 
participants should also refrain from sperm donation during this period.  If you think that you or 
your partner is pregnant, you should tell your study doctor or nurse at once.   
• Effective forms of birth control include: 
• Abstinence  
• intrauterine device (IUD) 
• hormonal [birth control pills, injections, or implants]  
• tubal ligation 
• vasectomy 
 
Gene Therapy Long Term Follow up (Retroviral Vectors)  
Because we do not know the long term side effects of gene therapy, we will collect your blood 
over the next several years, frequently at first and then less frequently.  If you return to your 
referring physician after treatment here we will ask you to have your physician send your blood 
specimens here for this testing.   This testing will determine if the cells have grown or changed in 
your body.  We will test your blood immediately after you receive the cells, and then at 3, 6 and 
12 months (2 teaspoons each time).  If all of the tests are normal and show no change, we will 
collect blood from you every year after that to store in case you develop symptoms later. According 
to FDA requirements, we need you to return annually to the NIH for a physical examination for 
five years after you receive the cells.  After that time, we will be sending you a questionnaire to 
get information regarding your health for the next ten years, for a total follow up time period of 15 
years.  For this reason, we ask that you continue to provide us with a current address and telephone 
number, even after you complete this research study.   At the time of your death, no matter the 
cause, we may request permission for an autopsy in order to obtain vital information concerning 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 7 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
the safety of this experimental treatment approach.  Please discuss this with your family to inform 
them of this request. 
 
Risks or Discomforts of Participation 
 
What side effects or risks can I expect from being in this study?  
The risks and discomforts of this research study can be significant.  This experimental treatment 
can lead to long-term decrease in your immune function.  It is also possible that you may lose your 
fertility following this experimental treatment. It is possible, although unlikely, that this 
experimental treatment may cause your death.  
 
We will discuss the side effects of this experimental treatment with you. You will be given 
medicines, transfusions, and treatments to prevent or treat the side effects including drugs to 
prevent and/or treat different types of infections.  We will try to make you as comfortable as 
possible.  You should talk to your study doctor about any symptoms that you experience while 
taking part in the study. 
 
Cell Infusion 
The cells we will be giving you have been modified with a retrovirus that has the gene for anti-
MAGE-A3-DP4 incorporated in the retrovirus.    The cells could cause you to develop another 
type of cancer, such as leukemia or lymphoma, although this has never been seen in any gene 
therapy trial involving lymphocytes.  These specific gene-modified cells have not been given to 
patients and thus we do not have much information about their side effects.  In a prior study using 
a different part of the MAGE gene, two patients of 9 who received this treatment died, probably 
due to the treatment.  We stopped that study but continued research to try to determine the cause 
of the side effects from the treatment.  The anti-MAGE-A3-DP4 is different from the MAGE used 
in the earlier trial and we do not anticipate that this anti-MAGE-A3-DP4 will cause the same side 
effects.  In addition, to using a different MAGE molecule, we are selecting a different kind of 
white blood cell to grow (CD4 instead of CD8). 
 
Potential risks include:  
• Fever, chills and shortness of breath, which may last for a few hours (common) 
• Lung congestion causing shortness of breath  
• Autoimmune reaction such as loss of skin pigment (known as vitiligo) or 
inflammation of the eye (uveitis) which may require the use of steroid eye drops.  
• As this is a new experimental therapy which has not been given to patients, side 
effects that we do not anticipate that may cause your condition to deteriorate may be 
encountered.  Any new information that becomes available during the course of this 
study will be shared with you. 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 8 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
• You will be treated on this gene transfer protocol with a viral vector that was 
manufactured at the NCI Surgery Branch Vector Production Facility before May 
2016.   An internal review of the facility that made the vector for this protocol 
determined that the facility needed to be closed due to manufacturing issues.  We 
know of no additional risks related to the previously produced vector for patients who 
have received cells with vectors made in this facility as the vectors were extensively 
tested by outside experts.  Therefore, the IRB has determined that the potential 
benefit to you outweighs the potential risks. 
 
Aldesleukin (IL-2)  
When IL-2 is given through an intravenous catheter, it can make you feel like you have the flu.  It 
can also cause confusion and mental status changes making you unable to make sound decisions.  
Prior to beginning treatment, we will ask you to complete a Durable Power of Attorney so that a 
person of your choosing can make health care decisions for you in case you develop these side 
effects.  In our experience giving IL-2 to over 2,000 patients we have found that these side effects 
go away within a few days of stopping the IL-2. 
Medications 
The side effects of cyclophosphamide, fludarabine, high dose aldesleukin and some of the other 
medications you will receive are listed below:  
Cyclophosphamide and Fludarabine side effects 
Common 
Less Common 
Rare 
▪ 
Changes in blood counts 
including: low red cell 
count (causing fatigue and 
shortness of breath), low 
platelet count (increasing 
the risk of bleeding and 
bruising), decrease in 
white blood cells 
(increasing the risk of 
infection and the need for 
treatment with antibiotics 
or other treatment)  
▪ 
Loss of appetite, nausea, 
vomiting,  
▪ 
Diarrhea, stomach pain 
▪ 
Mouth sores 
▪ 
Hair loss 
▪ 
Fatigue 
▪ 
Muscle or joint aches  
▪ 
Bleeding 
▪ 
Infection 
▪ 
Bladder irritation 
with bloody urine 
▪ 
Severe allergic 
reaction (difficulty 
breathing/swelling) 
▪ 
Headache or 
dizziness 
▪ 
Sweating 
▪ 
Swelling of arms 
or legs 
▪ 
Skin changes, rash, 
blisters  
▪ 
Weakness 
▪ 
Hearing loss 
▪ 
Heart damage 
▪ 
Lung damage 
▪ 
Kidney damage 
▪ 
Inflammation of the eye resulting in 
blindness 
▪ 
Inflammation of nervous system resulting 
in death 
▪ 
Epstein Barr Virus Lymphoma.  This can 
be fatal (Two patients on other studies in 
the Surgery Branch developed EBV 
lymphoma, and both died as a result of this 
disease.) 
▪ 
Loss of fertility 
▪ 
Death due to complications resulting from 
suppression of the immune function which 
resulted in a severe infection  


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 9 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
Aldesleukin side effects 
Common 
Less common  
Rare 
▪ 
Fever, chills, and 
fatigue   
▪ 
Lowered platelet and 
red blood cell levels 
that may require 
transfusions  
▪ 
Significant fluid 
retention causing 
weight gain (as much as 
20 pounds).  
▪ 
Low blood pressure 
▪ 
Increased heart rate 
▪ 
Low urine output 
▪ 
Swelling in your 
extremities,  
▪ 
Fluid in your lungs that 
can require oxygen  
▪ 
Dry mouth, nausea, 
vomiting and diarrhea; 
▪ 
Rash, itching; and 
changes in skin or hair 
pigmentation, called 
vitiligo;   
▪ 
Changes in mental 
status, including 
confusion, difficulty 
sleeping or vivid 
dreams; this can be 
severe and require 
sedation and monitoring 
in the ICU 
 
▪ 
Decrease in thyroid 
function that may 
require daily thyroid 
hormone replacement; 
▪ 
Abnormal kidney and 
liver function that can 
be severe;   
▪ 
Abnormal heartbeats or 
low blood pressure that 
may require treatment 
in the ICU.  
▪ 
Breathing problems 
which may need 
monitoring in ICU and 
insertion of a breathing 
tube.   
 
▪ 
Bowel perforation (a hole) 
requiring longer hospitalization 
or surgery. 
▪ 
Autoimmune disease, where 
your immune system attacks 
cells in organs of your body.  
Should this occur, you will be 
treated with steroids to stop the 
immune response. 
▪ 
Damage to the heart muscle or 
heart attack 
▪ 
Loss of blood flow to the 
extremities due to medicines 
used to treat very low blood 
pressure and shock. In one 
instance a patient had to have her 
lower arm amputated after 
treatment with these medicines. 
▪ 
IL-2 is mixed with human 
albumin which could cause an 
allergic reaction or potentially 
transmit viral infections, 
although we have not had this 
occur. 
 
 
 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 10 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
Support Medications – side effects 
Common 
Less common 
Rare 
Filgrastim (To increase production of white blood cells) 
▪ 
Bone Pain 
▪ 
Severe headache 
▪ 
Severe breathing problems 
▪ 
Rupture of your spleen 
Bactrim (To prevent a specific type of pneumonia) 
 
 
 
 
▪ 
Fever 
▪ 
Nausea, vomiting,  
▪ 
Skin rash with itching 
▪ 
reduced number of 
white blood cells 
▪ 
Allergic reaction 
 
 
 
 
Fluconazole: (To prevent fungal infections)  
▪ 
Headache 
▪ 
Nausea, vomiting, 
diarrhea, abdominal 
pain 
▪ 
Itching  
 
▪ 
A skin disorder called Stevens 
Johnson Syndrome, which can 
be fatal 
▪ 
Liver damage which may be 
permanent 
Acyclovir and Valacyclovir 
 
▪ 
Temporary decrease 
in kidney function 
which may not cause 
any symptoms 
▪ 
Nausea, vomiting, 
diarrhea, constipation  
▪ 
Pain and irritation at 
place of injection 
▪ 
Skin rash, hives, itching  
▪ 
Tremors, dizziness, Confusion, 
seizures 
▪ 
Fatigue  
▪ 
Blood in the urine 
 
Prior to and throughout this study you will undergo many tests to determine the size and extent of 
your tumor, as well as the impact of the treatment.  Multiple blood tests will be performed and 
some of your serum and lymphocytes will be stored for future testing.  Blood and tissue samples 
collected from you may be stored and used in the future to study scientific questions related to this 
protocol.  If there are any risks to you or your family associated with these future scientific studies 
which are not covered in this consent form, your consent will be obtained before such studies are 
performed. 
 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 11 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
If your disease progresses or recurs after this experimental treatment, then you will no longer 
receive treatment in this protocol, though you may be eligible to be considered for other protocols 
at the National Cancer Institute, NIH or referred elsewhere for treatment.  
 
Gene Therapy Risk of Cancer and Other Diseases  
We are unsure if this type of gene therapy will cause you to become sick in the future.  It is possible 
that it may cause your immune system or nerves not to work well or cause a sickness of your blood 
cells or even a cancer (for example leukemia).  We do not know if you will develop any of these 
disorders, but you need to be aware of this possible risk.  Children in France and England received 
gene therapy for a particular disease of the immune system.  Most of the children were cured, but 
5 children out of 22 later developed leukemia and one died.  Experts who looked at these cases 
thought that the gene therapy caused the leukemia in these children.  To monitor you for this risk 
we will be testing your blood 3 months after cell infusion, then at 6 and 12 months, and then 
annually thereafter.  If we find that the cells we have given you grow out of control, chemotherapy 
will be given to you to kill the cells, given their risk of causing leukemia or a second cancer. This 
testing will be conducted on our protocol 09-C-0161, “Follow up Protocol for Subjects Previously 
Enrolled in NCI Surgery Branch Studies” to which you will be enrolled following treatment. 
 
Potential Benefits of Participation 
Are there benefits to taking part in this study?  
The aim of this study is to see if this experimental treatment will cause your tumors to shrink.  We 
do not know if you will receive personal, medical benefit from taking part in this study.  These 
potential benefits could include shrinking of your tumor or lessening of your symptoms, such as 
pain, that are caused by the cancer.  Because there is not much information about the effect of this 
treatment on cancer, we do not know if you will benefit from taking part in this study, although 
the knowledge gained from this study may help others in the future who have cancer.   
 
Alternative Approaches or Treatments 
What other choices do I have if I do not take part in this study?  
Other options for treatment of your cancer include: 
• Taking part in another study  
• Getting treatment or care for your cancer without being in a study 
• Getting comfort care which is also called palliative care. This type of care helps reduce 
pain, tiredness, appetite problems, and other problems caused by cancer. It does not treat 
the cancer directly, but instead tries to improve how you feel. Comfort care tries to keep 
you as active and comfortable as possible. 
 
Please talk to your doctor about these and other options. 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 12 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
Research Subject’s Rights 
What are the costs of taking part in this study?  
If you choose to take part in the study, the following will apply, in keeping with the NIH policy: 
• 
You will receive study treatment at no charge to you. This may include 
surgery, medicines, laboratory testing, x-rays or scans done at the Clinical 
Center, National Institutes of Health (NIH), or arranged for you by the 
research team to be done outside the Clinical Center, NIH if the study 
related treatment is not available at the NIH.  
• 
There are limited funds available to cover the cost of some tests and 
procedures performed outside the Clinical Center, NIH.  You may have to 
pay for these costs if they are not covered by your insurance company. 
• 
Medicines that are not part of the study treatment will not be provided or 
paid for by the Clinical Center, NIH. 
• 
Once you have completed taking part in the study, medical care will no 
longer be provided by the Clinical Center, NIH. 
Will your medical information be kept private?  
We will do our best to make sure that the personal information in your medical record will be kept 
private.  However, we cannot guarantee total privacy.  Organizations that may look at and/or copy 
your medical records for research, quality assurance, and data analysis include: 
 
• The National Cancer Institute (NCI) and other government agencies, like the Food and 
Drug Administration (FDA), which are involved in keeping research safe for people. 
• National Cancer Institute Institutional Review Board 
• The Study Sponsor (Center for Cancer Research) or their agents 
• Kite Pharma 
A description of this clinical trial will be available on http://www.Clinicaltrials.gov, as required 
by U.S. Law.  This Web site will not include information that can identify you.  At most the Web 
site will include a summary of the results.  You can search this Web site at any time.  
Stopping Therapy  
Your doctor may decide to stop your therapy for the following reasons: 
• if he/she believes that it is in your best interest 
• if your disease comes back during treatment 
• if you become pregnant 
• if you have side effects from the treatment that your doctor thinks are too severe 
• if new information shows that another treatment would be better for you 


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 13 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
 
In this case, you will be informed of the reason therapy is being stopped. 
You can stop taking part in the study at any time. However, if you decide to stop taking part in the 
study, we would like you to talk to the study doctor and your regular doctor first. 
If you decide at any time to withdraw your consent to participate in the trial, we will not collect 
any additional medical information about you. However, according to FDA guidelines, 
information collected on you up to that point may still be provided to the Sponsor. If you withdraw 
your consent and leave the trial, any samples of yours that have been obtained for the study and 
stored at the NCI can be destroyed upon request. However, any samples and data generated from 
the samples that have already been distributed to other researchers or placed in the research 
databases cannot be recalled and destroyed. 
You should understand that this study involves research and that your participation is voluntary. 
Unexpected or unforeseeable side effects may also occur. Your participation in this protocol may 
be terminated without your consent if your physician feels that it would not be safe for you to 
continue.  Any significant new findings that relate to this protocol will be discussed with you. 
 
Conflict of Interest 
 
The National Institutes of Health (NIH) reviews NIH staff researchers at least yearly for conflicts 
of interest. This process is detailed in a Protocol Review Guide. You may ask your research team 
for a copy of the Protocol Review Guide or for more information.  Members of the research team 
who do not work for NIH are expected to follow these guidelines but they do not need to report 
their personal finances to the NIH. 
 
Members of the research team working on this study may have up to $15,000 of stock in the 
companies that make products used in this study.  This is allowed under federal rules and is not a 
conflict of interest.  
 
The National Institutes of Health and the research team for this study are working with Kite 
Pharma to see if this type of study could be done at institutions other than the NIH Clinical 
Center.  Kite Pharma also provides financial support for this study. 
 
Use of Specimens and Data for Future Research 
 
Blood and tissue collected during the course of this study will be used for future research and 
will be stored, tracked and disposed of under our companion protocol 03-C-0277, (Cell Harvest 
and Preparation for Surgery Branch Adoptive Cell Therapy Protocols) on which you have 
already been enrolled.   


MEDICAL RECORD 
CONTINUATION SHEET for either: 
NIH 2514-1, Consent to Participate in A Clinical Research Study 
NIH 2514-2, Minor Patient’s Assent to Participate In A Clinical Research Study 
STUDY NUMBER:  14-C-0052 
CONTINUATION: page 14 of 16 pages 
 
PATIENT IDENTIFICATION 
CONTINUATION SHEET for either: 
NIH-2514-1 (07-09) 
NIH-2514-2 (10-84) 
P.A.:  09-25-0099 
File in Section 4:  Protocol Consent 
 
 
 
In addition, to advance science, it is helpful for researchers to share information they get from 
studying human samples.  They do this by putting it into one or more scientific databases, where 
it is stored along with information from other studies. A researcher who wants to study the 
information must apply to the database and be approved.  Researchers use specimens and data 
stored in scientific databases to advance science and learn about health and disease.   
 
We plan to keep some of your specimens and data that we collect and use them for future 
research and share them with other researchers. We will not contact you to ask about each of 
these future uses.  These specimens and data will be stripped of identifiers such as name, address 
or account number, so that they may be used for future research on any topic and shared broadly 
for research purposes. Your specimens and data will be used for research purposes only and will 
not benefit you.  It is also possible that the stored specimens and data may never be used.  
Results of research done on your specimens and data will not be available to you or your doctor.  
It might help people who have cancer and other diseases in the future.   
 
If you do not want your stored specimens and data used for future research, please contact us in 
writing and let us know that you do not want us to use your specimens and/or data.  Then any 
specimens that have not already been used or shared will be destroyed and your data will not be 
used for future research. However, it may not be possible to withdraw or delete materials or data 
once they have been shared with other researchers. 
 
 


MEDICAL RECORD 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
• 
Adult Patient or 
• Parent, for Minor Patient 
STUDY NUMBER:   14-C-0052 
CONTINUATION: page 15 of 16 pages 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH 
STUDY (Continuation Sheet) 
• 
Adult Patient or 
• Parent, for Minor Patient 
NIH-2514-1 (07-09) 
P.A.:  09-25-0099 
 
File in Section 4:  Protocol Consent 
 
 
 
OTHER PERTINENT INFORMATION 
1. 
Confidentiality.  When results of an NIH research study are reported in medical journals 
or at scientific meetings, the people who take part are not named and identified.  In most cases, the 
NIH will not release any information about your research involvement without your written 
permission.  However, if you sign a release of information form, for example, for an insurance 
company, the NIH will give the insurance company information from your medical record.  This 
information might affect (either favorably or unfavorably) the willingness of the insurance 
company to sell you insurance. 
The Federal Privacy Act protects the confidentiality of your NIH medical records.  However, you 
should know that the Act allows release of some information from your medical record without 
your permission, for example, if it is required by the Food and Drug Administration (FDA), 
members of Congress, law enforcement officials, or authorized hospital accreditation 
organizations. 
2. 
Policy Regarding Research-Related Injuries.  The Clinical Center will provide short-
term medical care for any injury resulting from your participation in research here.  In general, no 
long-term medical care or financial compensation for research-related injuries will be provided by 
the National Institutes of Health, the Clinical Center, or the Federal Government.  However, you 
have the right to pursue legal remedy if you believe that your injury justifies such action. 
3. 
Payments.  The amount of payment to research volunteers is guided by the National 
Institutes of Health policies.  In general, patients are not paid for taking part in research studies at 
the National Institutes of Health.  Reimbursement of travel and subsistence will be offered 
consistent with NIH guidelines. 
4. 
Problems or Questions.  If you have any problems or questions about this study, or about 
your rights as a research participant, or about any research-related injury, contact the Principal 
Investigator, Steven A. Rosenberg, M.D., Ph.D., Building 10, Room 3-3940, Telephone:  240-
760-6218. If you have any questions about the use of your specimens or data for future research 
studies, you may also contact the Office of the Clinical Director, Telephone:  240-760-6070. You 
may also call the Clinical Center Patient Representative at 301-496-2626.  
5. 
Consent Document.  Please keep a copy of this document in case you want to read it again. 
 
 


MEDICAL RECORD 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH STUDY 
• 
Adult Patient or 
• Parent, for Minor Patient 
STUDY NUMBER:   14-C-0052 
CONTINUATION: page 16 of 16 pages 
 
PATIENT IDENTIFICATION 
CONSENT TO PARTICIPATE IN A CLINICAL RESEARCH 
STUDY (Continuation Sheet) 
• 
Adult Patient or 
• Parent, for Minor Patient 
NIH-2514-1 (07-09) 
P.A.:  09-25-0099 
 
File in Section 4:  Protocol Consent 
 
 
 
 
COMPLETE APPROPRIATE ITEM(S) BELOW: 
A. Adult Patient’s Consent 
B. Parent’s Permission for Minor Patient. 
I have read the explanation about this study 
and have been given the opportunity to 
discuss it and to ask questions.  I hereby 
consent to take part in this study. 
I have read the explanation about this study 
and have been given the opportunity to 
discuss it and to ask questions.  I hereby give 
permission for my child to take part in this 
study.   
(Attach NIH 2514-2, Minor’s Assent, if 
applicable.) 
 
  
  
 
 
 
Signature of Adult Patient/ 
Legal Representative 
Date 
Signature of Parent(s)/ 
Guardian 
Date 
 
 
Print Name 
 
 
Print Name 
C. Child’s Verbal Assent (If Applicable) 
The information in the above consent was described to my child and my child agrees to 
participate in the study. 
 
 
 
 
 
 
 
Signature of Parent(s)/Guardian 
Date 
Print Name 
THIS CONSENT DOCUMENT HAS BEEN APPROVED FOR USE  
FROM JUNE 24, 2019 THROUGH JULY 08, 2020.  
 
 
 
 
 
 
 
 
 
 
 
 
Signature of Investigator 
Date 
Signature of Witness 
Date 
 
 
Print Name 
 
 
Print Name