- MART-1 is a gene present in melanoma cells.
- An experimental procedure developed for treating patients with melanoma uses the
anti-MART-1 F5 gene and a type of virus to make special cells called anti-MART-1 F5
cells that are designed to destroy the patient's tumor. These cells are created in the
laboratory using the patient's own tumor cells or blood cells.
- The treatment procedure also uses a vaccine called ALVAC MART-1, made from a virus that
ordinarily infects canaries and is modified to carry a copy of the MART-1 gene. The
virus cannot reproduce in mammals, so it cannot cause disease in humans. When the
vaccine is injected into a patient, it stimulates cells in the immune system that may
increase the efficiency of the anti-MART-1 F5 cells.
-To evaluate the safety and effectiveness of anti-MART-1 F5 and the ALVAC vaccine in
treating patients with advanced melanoma.
-Patients 18 years of age with metastatic melanoma for whom standard treatments have not
- Patients undergo scans, x-rays and other tests and leukapheresis to obtain white cells
for laboratory treatment.
- Patients have 7 days of chemotherapy to prepare the immune system for receiving the
- Patients receive the ALVAC vaccine, anti-MART-1 F5 cells and IL-2 (an approved
treatment for advanced melanoma). The anti-MART-1 F5 cells are given as an infusion
through a vein. The vaccine is given as injections just before the infusion of
anti-MART-1 F5 cells and again 2 weeks later. IL-2 is given as a 15-minute infusion
every 8 hours for up to 5 days after the cell infusion for a maximum of 15 doses.
- After hospital discharge, patients return to the clinic for periodic follow-up with a
physical examination, review of treatment side effects, laboratory tests and scans
every 1 to 6 months....
- We have engineered human PBLs to express an anti-MART-1 T-cell receptor that recognizes
an HLA-A 0201 restricted epitope derived from the TIL clone DMF5.
- We constructed a single retroviral vector that contains both alpha and infinity chains
and can mediate genetic transfer of this 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 IFN-(but no significant secretion was observed in
control co-cultures with cell lines.
- The anti-MART-1 F5 TCR transduced 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).
-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.
- 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.
- 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 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
- Patients will undergo complete evaluation of tumor with physical examination, 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
- 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 PR plus CR rate (p1=0.20).
- INCLUSION CRITERIA:
1. Metastatic melanoma with measurable disease.
2. Previously received high dose IL-2 and have been either non-responders
(progressive disease) or have recurred.
3. Positive for MART-1 by IHC which will be reviewed by the Laboratory of Pathology
4. 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 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 HLA-A 0201 positive.
- 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 hepatitis C antibody unless antigen
- Absolute neutrophil count greater than 1000/mm(3) without the support of filgrastim.
- WBC (greater than 3000/mm(3)).
- Platelet count greater than 100,000/mm(3).
- Hemoglobin greater than 8.0 g/dl.
- Serum ALT/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.
o. 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).
p. Six weeks must have elapsed since prior MDX-010 therapy to allow antibody levels
q. 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
7. History of coronary revascularization or ischemic symptoms.
8. Any patient known to have an 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
- Age greater than or equal to 60 years old.
10. Documented 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.