Drosophila-generated CTL

Background:

- Adoptive transfer studies in patients with metastatic melanoma following
lymphodepletion have resulted in up to 50% objective response rates with a 10-15% rate
of complete responses.

- A novel method involves the use of insect cell lines which do not express any native
major histocompatibility complex (MHC) molecules.

- When stably transfected with human MHC molecules and appropriate adhesion and
costimulatory molecules, a Drosophila cell line can potently stimulate tumor-reactivity
in vitro from human peripheral blood lymphocytes (PBL).

- The current proposed transfer of Drosophila-cell stimulated autologous cluster of
differentiation 8 (CD8) plus PBL administered in conjunction with a lymphodepleting
preparative regimen, with or without low-dose aldesleukin would represent a
significantly novel approach to adoptive immunotherapy.

Objectives:

- To determine whether infusion of CD8+ autologous PBL sensitized in vitro with peptide
pulsed HLA-A2-expressing Drosophila cells (CTL-05) and administered in combination with
a lymphodepleting preparative regimen and supportive systemic aldesleukin can result in
clinical tumor regression in human leukocyte antigen serotype within HLA-A A serotype
group (HLA-A2+) patients with metastatic melanoma.

- To determine the safety of the above regimen.

- To investigate the contribution of low-dose systemic aldesleukin to cell efficacy.

Eligibility:

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

- metastatic melanoma with measurable disease

- been previously treated with aldesleukin for melanoma;

- normal basic laboratory values.

Patients may not have:

- concurrent major medical illnesses;

- any form of primary or secondary immunodeficiency;

- requirement for systemic steroid therapy

Design:

- The first 20 patients enrolled (cohort 0) will receive a non-myeloablative lymphocyte
depleting preparative regimen followed by administration of intravenous CTL-05 and
low-dose subcutaneous aldesleukin (daily for 5 days).

- If 3 or more of the 20 patients respond, subsequent patients will be randomized between
two cohorts. Patients in cohort 1 will receive a non-myeloablative lymphocyte depleting
preparative regimen followed by administration of CTL-05 and low-dose subcutaneous
aldesleukin (daily for 5 days). Patients in Cohort 2 will receive a non-myeloablative
lymphocyte depleting preparative regimen followed by administration of CTL-05 and NO
subsequent aldesleukin.

- A complete evaluation will be conducted 8 weeks (plus or minus 2 weeks) after the
initiation of chemotherapy. The trial will be conducted using a small Simon MinMax
Phase II design in the initial phase and a Simon optimal design in the second phase. A
maximum of 35 patients may be accrued to each of cohorts 1 and 2. If no responses are
seen in the first 13 patients receiving no systemic aldesleukin, then accrual to that
cohort will cease. Total enrollment may be up to 90 patients.

- INCLUSION CRITERIA:

1. Metastatic cutaneous melanoma with measurable disease by Response Evaluation
Criteria in Solid Tumors (RECIST) criteria

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

3. Patients with 3 or less brain metastases are eligible. Note: If lesions are
symptomatic or greater than or equal to 1 cm each, these lesions must have been
treated and stable for 3 months for the patient to be eligible.

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

5. Able to understand and sign the Informed Consent Document

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

7. Life expectancy of greater than three months.

8. Patients of both genders must be willing to practice a highly effective method
of birth control during and for four months following treatment

9. Patients must be HLA-A*0201 positive

10. Serology:

1. 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 toxicities.)

2. Seronegative for hepatitis B antigen and hepatitis C antibody unless
antigen negative.

11. Hematology:

1. Absolute neutrophil count greater than 1000/mm^3 without the support of
filgrastim.

2. White blood cell (WBC) (> 3000/mm^3).

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

4. Hemoglobin greater than 8.0 g/dl.

12. Chemistry:

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

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

3. Total bilirubin less than or equal to 1.5 mg/dl, except in patients with
Gilbert's Syndrome who must have a total bilirubin less than 3.0 mg/dl.

13. 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 hypothyroidism, alopecia, or
vitiligo).

14. Six weeks must have elapsed since prior anti-CTLA4 antibody therapy to allow
antibody levels to decline.

15. Patients who have previously received anti-CTLA4 antibody must have a normal
colonoscopy with normal colonic biopsies.

EXCLUSION CRITERIA:

1. Active systemic infections, coagulation disorders or other active major medical
illnesses.

2. Any form of primary immunodeficiency (such as Severe Combined Immunodeficiency
Disease).

3. 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).

4. Requirement for systemic steroid therapy

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

6. History of coronary revascularization or ischemic symptoms

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

8. Documented LVEF of less than or equal to 45% tested in patients with:

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

2. Age greater than or equal to 60 years old

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

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

2. Symptoms of respiratory dysfunction

10. Pregnant or nursing women