We will prepare 1-butanol extracts of fresh human melanoma cells to use as an allogeneic """"""""vaccine"""""""" for active specific immunotherapy. The melanoma-associated antigens in each extract will be detected by a panel of mouse and human monoclonal antibodies and quantitated by a binding inhibition procedure we have developed. Human monoclonal antibodies will be continually developed from human-mouse and human-human fusions, with regional lymph node cells from melanoma patients as one partner and a nonsecretory mouse (M5) myeloma or a human B or myeloma cell line as the other partner. These human monoclonal antibodies will enable us to determine the presence of epitopes recognized by patients as """"""""foreign"""""""", in the butanol extracts used as immunogens. The toxicity and immunological effects of a standard pool of 3 antigenic extracts will be tested in a formal Phase I trial, with metastatic melanoma patients. Two-fold escalations will be performed in each group for a total of 3 levels of dosage, with and without low-dose cyclophosphamide as a potentiator of immunity. Besides noting standard toxicity, we will also measure suppressor T cells by various complementary methods, to determine the possible occurence of this potentially harmful consequence of immunization. Several tests for antibodies to melanoma including immune adherence, protein A mixed hemadsorption and two tests for cytophilic antibodies (LAI and ADCC), will be performed. We have also developed a new titration method based on competitive inhibition of monoclonal antibody binding, with a sensitivity of 1-2 ng of antibody, to measure most sensitively the serum antibody response in the patients. Delayed hypersensitivity will be studied by skin testing with the pool of melanoma extracts. We will also explore limiting dilution analysis of cultivated T cells as a new means of detecting cytolytic antimelanoma T cells, since other tests of cell-mediated immunity have been inconsistently reliable. The developmental aspects of this proposal, with improvement of competitive binding, limiting dilution analysis and suppressor cell methods, in fact constitute an important rationale for the entire study. This closely monitored clinical trial may help to define the most important antigens to include in a """"""""vaccine"""""""", leading to their selective use in future studies of specific active immunotherapy for melanoma.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA036233-03
Application #
3173744
Study Section
Experimental Immunology Study Section (EI)
Project Start
1984-01-01
Project End
1987-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Southern California
Department
Type
Schools of Medicine
DUNS #
041544081
City
Los Angeles
State
CA
Country
United States
Zip Code
90033
Mitchell, M S; Liggett, P E; Green, R L et al. (1994) Sustained regression of a primary choroidal melanoma under the influence of a therapeutic melanoma vaccine. J Clin Oncol 12:396-401
Mitchell, M S; Jakowatz, J; Harel, W et al. (1994) Increased effectiveness of interferon alfa-2b following active specific immunotherapy for melanoma. J Clin Oncol 12:402-11
Quan Jr, W D; Mitchell, M S (1993) Immunology and immunotherapy of melanoma. Cancer Treat Res 65:257-77
Kan-Mitchell, J; Huang, X Q; Steinman, L et al. (1993) Clonal analysis of in vivo activated CD8+ cytotoxic T lymphocytes from a melanoma patient responsive to active specific immunotherapy. Cancer Immunol Immunother 37:15-25
Mitchell, M S; Harel, W; Kan-Mitchell, J et al. (1993) Active specific immunotherapy of melanoma with allogeneic cell lysates. Rationale, results, and possible mechanisms of action. Ann N Y Acad Sci 690:153-66
LeMay, L G; Kan-Mitchell, J; Goedegebuure, P et al. (1993) Detection of melanoma-reactive CD4+ HLA-class I-restricted cytotoxic T cell clones with long-term assay and pretreatment of targets with interferon-gamma. Cancer Immunol Immunother 37:187-94
Mitchell, M S (1992) Chemotherapy in combination with biomodulation: a 5-year experience with cyclophosphamide and interleukin-2. Semin Oncol 19:80-7
Mitchell, M S (1992) Biomodulators in cancer treatment. J Clin Pharmacol 32:2-9
Mitchell, M S (1992) Principles of combining biomodulators with cytotoxic agents in vivo. Semin Oncol 19:51-6
Mitchell, M S; Harel, W; Groshen, S (1992) Association of HLA phenotype with response to active specific immunotherapy of melanoma. J Clin Oncol 10:1158-64

Showing the most recent 10 out of 28 publications