Project Identifier: LLNL-94-101
Project Title:
Immunological Treatment of Metastatic Melanoma
Principle Investigator:
Dr. Max W. Biggs
Project started in: 1994
Project Funding Information:
Project received funding in Fiscal Year 1995.
Project used human subjects in Fiscal Year 1995.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: January 18, 1995
IRB Approval Number: 94-101
Number of Human Subjects in the Last Reporting Period for this Project: 6
(Reporting periods vary.)
Type of Human Subjects Involvement:
STUDY AIM/PURPOSE:
The purpose of this study is to see whether the "soluble" antigens of an enteroviral autologous melanoma lysate, when injected into metastatic melanoma and followed by cyclophosphamide, will produce a remission in the treated metastatic lesions.
BACKGROUND:
There is extensive evidence in immunology that a normal suppressor branch of the system exists to limit the magnitude and duration of immune responses. It is postulated that "melanoma vaccines" fail because the host is tolerized to melanoma antigens.
In the past we have studied the effects of injection of a murine tumor lysate prepared in tissue culture by infection with a murine enterovirus. This was followed by the administration of cyclophosphamide. Both allogeneic and autologous mammary tumors in mice were studied. Ninety percent of the transplanted tumors were cured and significant remissions were obtained in 50% of the spontaneous tumors. Since the "vaccine" was filtered through a 50 nm Millipore filter and contained "soluble antigens" without aggregates and the cyclophosphamide was given AFTER the antigen, the mechanism of action is thought to be reversal of immune suppression. We believe that the cells involved with in vivo immune suppression were stimulated to divide and were then destroyed with cyclophosphamide.
FY'95 STUDIES:
In FY'95 we have studied 6 patients with metastatic melanoma to evaluate whether or not the murine experiments can be repeated in man. Melanoma tissue cultures have been established from these patients. We have demonstrated that human melanoma cells in culture can be infected and lysed with the enterovirus, poliovirus 1 (Sabin).
In 2 of the studied patients it has been possible to generate in vitro a cytotoxic T-cell lymphocyte response against their autologous melanoma cells using peripheral blood lymphocytes. Cr51 microcytotoxicity assays were used to make these determinations. We have demonstrated in these 2 patients that a melanoma lysate prepared with poliovirus 1 (Sabin) and filtered through a 50 nm Millipore filter is immunosuppressive and prevents the in vitro generation of a cytotoxic response. Thus we are now ready to repeat the animal experiments in man. This will be done in FY'96.
PLANNED FY'96 STUDIES:
PATIENT ELIGIBILITY: The patients must have multiple cutaneous or subcutaneous melanoma metastases. The patient must be immunologically competent.
TREATMENT PLAN: Several of the multiple metastatic lesions will be removed surgically. The pathology and character of the tumor infiltrating lymphocytes will be described. Melanoma cell tissue cultures will be established and the poliovirus lysate will be generated.
Several of the patient's remaining metastases will be injected with the autologous poliovirus lysate followed in 24 hours with a single dose of cyclophosphamide, 300 mg/M2. Changes in the treated tumors will be followed clinically. Any evidence of tumor regression will be recorded.
In addition some of the treated lesions will be removed surgically for evaluation and for comparison with pre-treatment biopsy data. The number and nature of the tumor infiltrating lymphocytes will be evaluated. The secretion of Interleukin-10 will be measured using RT-PCR methodologies.
If any evidence of a cytotoxic T-cell response in the treated tumors can be demonstrated, it is anticipated that such reversal of immune suppression will aid in the immunotherapy of metastatic melanoma with various tumor vaccines.