In active specific immunotherapy for human cancer, the vehicle for immunogenic stimulus, or the vaccine, has comprised tumor cells themselves, in one form or another. Tumor cells, however, might not be able to provide the antigenic and the co-stimulatory signals, two obligate necessities for T cell activation. In a search for a more effective vehicle for delivery of both signals, we seek to use the macrophage, a professional antigen presenting cell, to develop an autologous macrophage-based melanoma vaccine. Melanoma is an ideal tumor because it is an immunogenic tumor and the melanoma antigen gene has been structurally defined and the antigenic peptide sequenced. Accordingly, we propose to test the hypothesis that the state of immunologic unresponsiveness of hosts against autologous melanomas can be broken through a novel approach using active specific immunotherapy. The approach will incorporate a mechanism of delivery of the antigenic as well as the co-stimulatory signals, simultaneously, through various routes. Autologous macrophages will be pulsed with relevant tumor antigen derived peptides, or made to take up, process and present tumor associated antigens, or made to present the tumor antigen derived peptide following the introduction of the gene encoding the antigen into such APC by in vitro gene transfer technique. For this purpose, large numbers of macrophages will be grown from melanoma patients, and the cultured macrophages will be pulsed with the melanoma antigen (MAGE-1) peptide EADPTGHSY or will be loaded (through phagocytosis or endocytosis) with the autologous melanoma lysates for processing and presenting of antigen fragments on their MHC molecules. Patients, who are at risk for recurrence or who have recurrent disease and who are HLA A1/MAGE1 positive, will be vaccinated with the peptide pulsed Mphi. Other patients will be treated with autologous Mphi loaded with lysates of autologous melanomas as sources for melanoma associated protein antigen. All patients will be first primed, intravenously, with the peptide pulsed or the antigen loaded Mphi, followed by four intradermal challenge doses. A method of skin testing with the antigen or the vaccine will be developed to assess the immunogenicity of the vaccine. In addition, extensive laboratory evaluation of the immune status of the vaccinated patients will be undertaken to answer the question of whether or not this type of immunization can induce a T cell mediated response in vivo. T cells isolated from immunization sites, skin test sites and from metastatic deposits will be expanded in vitro and cloned for detailed phenotypic and functional analyses (proliferative and cytokine synthetic ability upon stimulation with the immunogen, cytolytic capacity against the autologous and other targets). Finally, we shall explore the feasibility of transferring the MAGE-1 gene along with the co-stimulatory molecule B7 into Mphi by in vitro gene transfer technique for future in vivo testing of such genetically altered Mphi vaccine. Results of in vitro assays for immunogenicity, skin testing and therapeutic result, if any, will be analyzed to establish clinical correlates. These studies will facilitate the development of a unique and effective form of specific biologic cancer therapy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01CA061398-02
Application #
2102136
Study Section
Special Emphasis Panel (SRC (59))
Project Start
1993-09-01
Project End
1997-06-30
Budget Start
1994-09-01
Budget End
1995-06-30
Support Year
2
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Connecticut
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Farmington
State
CT
Country
United States
Zip Code
06030
Chhabra, Arvind; Mehrotra, Shikhar; Chakraborty, Nitya G et al. (2006) Activation-induced cell death of human melanoma specific cytotoxic T lymphocytes is mediated by apoptosis-inducing factor. Eur J Immunol 36:3167-74
Chakraborty, Nitya G; Chattopadhyay, Subhasis; Mehrotra, Shikhar et al. (2004) Regulatory T-cell response and tumor vaccine-induced cytotoxic T lymphocytes in human melanoma. Hum Immunol 65:794-802
Mehrotra, Shikhar; Chhabra, Arvind; Chakraborty, Abolokita et al. (2004) Antigen presentation by MART-1 adenovirus-transduced interleukin-10-polarized human monocyte-derived dendritic cells. Immunology 113:472-81
Mehrotra, Shikhar; Chhabra, Arvind; Chattopadhyay, Subhasis et al. (2004) Rescuing melanoma epitope-specific cytolytic T lymphocytes from activation-induced cell death, by SP600125, an inhibitor of JNK: implications in cancer immunotherapy. J Immunol 173:6017-24
Mehrotra, Shikhar; Stevens, Robert; Zengou, Ryan et al. (2003) Regulation of melanoma epitope-specific cytolytic T lymphocyte response by immature and activated dendritic cells, in vitro. Cancer Res 63:5607-14
Chakraborty, A; Li, L; Chakraborty, N G et al. (2000) Stimulatory and inhibitory differentiation of human myeloid dendritic cells. Clin Immunol 94:88-98
Chakraborty, N G; Li, L; Sporn, J R et al. (1999) Emergence of regulatory CD4+ T cell response to repetitive stimulation with antigen-presenting cells in vitro: implications in designing antigen-presenting cell-based tumor vaccines. J Immunol 162:5576-83
Chakraborty, A; Li, L; Chakraborty, N G et al. (1999) Stimulatory and inhibitory maturation of human macrophage-derived dendritic cells. Pathobiology 67:282-6
Chakraborty, N G; Sporn, J R; Tortora, A F et al. (1998) Immunization with a tumor-cell-lysate-loaded autologous-antigen-presenting-cell-based vaccine in melanoma. Cancer Immunol Immunother 47:58-64
Perez-Diez, A; Butterfield, L H; Li, L et al. (1998) Generation of CD8+ and CD4+ T-cell response to dendritic cells genetically engineered to express the MART-1/Melan-A gene. Cancer Res 58:5305-9

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