| Project/Area Number |
04454287
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | Kobo University |
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Principal Investigator |
HAYASHIBE Kazuhito Kobe University., Medicine, Assistant Professor, 医学部・付属病院, 講師 (40198875)
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| Co-Investigator(Kenkyū-buntansha) |
HOTTA Hakku Kobe University., Medicine, Professor, 医学部, 教授 (40116249)
ICHIHASHI Masamitsu Kobe University., Medicine, Professor, 医学部, 教授 (00030867)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1993: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1992: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | Melanoma / Immunotherapy / Vaccine / Melanoma Antigen / Gene Technology / Recombinant Protein |
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| Research Abstract |
This study aimed at the identification and characterization of human melanoma-associated antigens immunogenic in patients. The cloned cDNA encoding 34 Kd peptide antigen showed no significant homology of nucleic acid sequence with previously reported mammalian genes. The clone D-1 cDNA hybridized with mRNA from cultured human melanomas, neuroblastoma, erythroleukemia and lympoma cells but not with mRNA from normal fibroblasts, lymphocytes and renal carcinoma cells. The tissue distribution of mRNA corresponding to D-1 was analyzed by in-situ hybridization utilizing D-1 cRNA prove, and showed to be expressed on melanoma cells in vivo. No significant expression on major normal organs and other skin cancers was detected. Murine polyclonal anti-D-1 antibodies were raised against Balb/c mice with immunization of recombinant D-1 peptide and showed the invivo localization of D-1 antigen on the surface membrane of melanoma cells in an almost identical fashion to in-situ hybridization. But the strong expression of D-1 antigen in the cytoplasm of some melanoma tissues indicated the need of HLA molecules with certain subclass to express D-1 antigen on the surface. The activated lymphocytes in vitro with D-1 recombinant peptide showed substantial cytotoxic activities to autologous melanoma cells comparing with the control cells K562 in co-culture for 28 days. Active, specific immunotherapy with D-1 recombinant peptide vaccine is in process on patients with StageIV melanoma. Serological observation of anti-D-1 antisera in patients received vaccination revealed two groups with high and low titer. The comparative analysis of HLAclass I type between these groups is planned.
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