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Phase II Study of Metastatic Melanoma Using Lymphodepleting Conditioning Followed by Infusion of Anti-MART-1 F5 TCR-Gene Engineered Lymphocytes and ALVAC Virus Immunization

Phase 2
18 Years
Not Enrolling
Metastatic Melanoma, Skin Cancer

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Trial Information

Phase II Study of Metastatic Melanoma Using Lymphodepleting Conditioning Followed by Infusion of Anti-MART-1 F5 TCR-Gene Engineered Lymphocytes and ALVAC Virus Immunization


- We have engineered human PBLs to express an anti-MART-1 T-cell receptor that recognizes
an human leukocyte antigens (HLA-A) 0201 restricted epitope derived from the tumor
infiltrating lymphocytes (TIL) clone DMF5.

- We constructed a single retroviral vector that contains both alpha and infinity chains
and can mediate genetic transfer of this T cell receptor (TCR) with high efficiency
without the need to perform any selection.

- In co-cultures with HLA-A 0201 positive melanoma, anti-MART-1 F5 TCR transduced T cells
secreted significant amount of interferon (IFN)-(but no significant secretion was
observed in control co-cultures with cell lines.

- The anti-MART-1 F5 TCR transduced peripheral blood lymphocytes (PBL) could efficiently
kill HLA-A 0201 positive tumors. There was little or no recognition of normal
fibroblasts cells.

- This TCR is over 10 times more reactive with melanoma cells than the MART-1 TCR that
mediated tumor regression in two patients with metastatic melanoma.

- In this trial we would like to test our hypothesis that the addition of an anti-tumor
ALVAC vaccine will result in clinical tumor regression and persistence of the
transferred cells (as is the case in murine models).


Primary objectives:

-Determine if the administration of anti-MART-1 F5 TCR -engineered peripheral blood
lymphocytes, ALVAC anti-tumor immunization, and aldesleukin to patients following a
nonmyeloablative but lymphoid depleting preparative regimen will result in clinical tumor
regression in patients with metastatic melanoma.

Secondary objectives:

- Determine the in vivo survival of TCR gene-engineered cells.

- Determine the toxicity profile of this treatment regimen.


Patients who are HLA-A 0201 positive and 18 years of age or older must have:

- metastatic melanoma;

- previously received and have been a non-responder to or recurred after aldesleukin;

- normal values for basic laboratory values.

Patients may not have:

- concurrent major medical illnesses;

- any form of primary or secondary immunodeficiency;

- severe hypersensitivity to any of the agents used in this study;

- contraindications for high dose aldesleukin administration.

- Design:

- peripheral blood mononuclear cells (PBMC) obtained by leukapheresis (approximately 5
times 10^9 cells) will be cultured in the presence of anti-CD3 (OKT3) and aldesleukin
in order to stimulate T-cell growth.

- Transduction is initiated by exposure of approximately 10^8 to 5 X 10^8 cells to
retroviral vector supernatant containing the anti-MART-1 F5 TCR genes. These transduced
cells will be expanded and tested for their anti-tumor activity.

- Once engineered PBMC are demonstrated to be biologically active according to the
strict-criteria outlined in the Certificate of Analysis, 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 PBMC plus intravenous (IV) aldesleukin (720,000 IU/kg q8h
for a maximum of 15 doses). Approximately 2 hours prior to cell infusion, patients will
be immunized with ALVAC virus expressing the tumor antigen. ALVAC immunization will be
repeated at 2 weeks.

- Patients will undergo complete evaluation of tumor with physical examination, computed
tomography (CT) of the chest, abdomen and pelvis and clinical laboratory evaluation
four to six weeks after treatment and then monthly for approximately 3 to 4 months or
until off study criteria are met.

- The study will be conducted using a phase II optimal design where 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 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.

- The objective will be to determine if the combination of high dose aldesleukin,
lymphocyte depleting chemotherapy, ALVAC immunization and anti-MART-1 F5 TCR-gene
engineered lymphocytes is able to be associated with a clinical response rate that can
rule out 5 percent (p0=0.05) in favor of a modest 20 percent partial response (PR) plus
complete response (CR) rate (p1=0.20).

Inclusion Criteria


1. Metastatic melanoma with measurable disease.

2. Previously received high dose interleukin-2 (IL-2) and have been either
non-responders (progressive disease) or have recurred.

3. Positive for MART-1 by immunohistochemistry (IHC) which will be reviewed by the
Laboratory of Pathology at National Cancer Institute (NCI).

4. Tumor infiltrating lymphocytes (TIL) cells not available for treatment on other
Surgery Branch protocols.

5. Greater than or equal to 18 years of age.

6. Willing to sign a durable power of attorney.

7. Able to understand and sign the Informed Consent Document.

8. Clinical performance status of Eastern Cooperative Oncology Group (ECOG) 0 or 1.

9. Life expectancy of greater than three months.

10. Patients of both genders must be willing to practice birth control for four months
after receiving the preparative regimen.

11. Patients must be human leukocyte antigens (HLA-A) 0201 positive.

12. Serology:

- Seronegative for human immunodeficiency virus (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

- Seronegative for hepatitis B antigen and hepatitis C antibody unless antigen

13. Hematology:

- Absolute neutrophil count greater than 1000/mm^3 without the support of

- White blood cell (WBC) (greater than 3000/mm^3.

- Platelet count greater than 100,000/mm^3.

- Hemoglobin greater than 8.0 g/dl.

14. Chemistry:

- Serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) less or
equal to 2.5 times the upper limit of normal.

- Serum creatinine less than or equal to 1.6 mg/dl.

- Total bilirubin less than or equal to 2.0 mg/dl, except in patients with
Gilbert's Syndrome who must have a total bilirubin less than 3.0 mg/dl.

15. 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).

16. Six weeks must have elapsed since prior MDX-010 therapy to allow antibody levels to

17. Patients who have previously received MDX-010 or ticilimumab must have a normal
colonoscopy with normal colonic biopsies.


1. Women of child-bearing potential who are pregnant or breastfeeding because of the
potentially dangerous effects of the preparative chemotherapy on the fetus or infant.

2. Active systemic infections, coagulation disorders or other major medical illnesses of
the cardiovascular, respiratory or immune system, myocardial infarction, cardiac
arrhythmias, obstructive or restrictive pulmonary disease.

3. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency

4. Ongoing 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).

5. Systemic steroid therapy.

6. History of severe immediate hypersensitivity reaction to any of the agents used in
this study.

7. History of coronary revascularization or ischemic symptoms.

8. Any patient known to have an left ventricular ejection fraction (LVEF) less than or
equal to 45 percent.

9. Documented LVEF of less than or equal to 45 percent tested in patients with:

- Clinically significant atrial and/or ventricular arrhythmias including but not
limited to: atrial fibrillation, ventricular tachycardia, second or third degree
heart block.

- Age greater than or equal to 60 years old.

10. Documented forced expiratory volume in 1 second (FEV1) less than or equal to 60
percent predicted tested in patients with:

- A prolonged history of cigarette smoking (20 pk/yrs of smoking).

- Symptoms of respiratory dysfunction.

Type of Study:


Study Design:

Allocation: Non-Randomized, Endpoint Classification: Efficacy Study, Intervention Model: Single Group Assignment, Masking: Open Label, Primary Purpose: Treatment

Outcome Measure:

Number of Participants With Metastatic Melanoma Who Develop Clinical Tumor Regression (CR or PR)

Outcome Description:

Clinical tumor response is assessed by the Response Evaluation Criteria in Solid Tumors (RECIST) v.1.0 criteria. Complete response (CR) is a disappearance of all target lesions. Partial response (PR) is a 30% decrease in lesions taking as reference the baseline sum longest diameter (LD). For details about the RECIST criteria see the protocol link module.

Outcome Time Frame:

4-6 weeks after treatment and then monthly for approximately 3 to 4 months or until off study criteria are met

Safety Issue:


Principal Investigator

Steven A Rosenberg, M.D.

Investigator Role:

Principal Investigator

Investigator Affiliation:

National Cancer Institute, National Institutes of Health


United States: Federal Government

Study ID:




Start Date:

January 2008

Completion Date:

March 2011

Related Keywords:

  • Metastatic Melanoma
  • Skin Cancer
  • Immunotherapy
  • Metastatic Melanoma
  • Vaccination
  • Tumor Regression
  • Skin Cancer
  • Skin Neoplasms
  • Melanoma



National Institutes of Health Clinical Center, 9000 Rockville PikeBethesda, Maryland  20892