| Project/Area Number |
04671070
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | DEPARTMENT OF OPHTHALMOLOGY, SHINSHU UNIVERSITY |
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Principal Investigator |
NOROSE Kazumi DEPARTMENT OF OPHTHALMOLOGY, SHINSHU UNIVERSITY SCHOOL OF MEDICINE LECTURER, 医学部・附属病院・眼科, 講師 (30156244)
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| Co-Investigator(Kenkyū-buntansha) |
SEGAWA Katsuzo DEPARTMENT OF OPHTHALMOLOGY, SHINSHU UNIVERSITY SCHOOL OF MEDICINE PROFESSOR, 医学部・眼科学教室, 教授 (70020857)
YANO Akihiko DEPARTMENT OF MEDICAL ZOOLOGY, NAGASAKI UNIVERSITY SCHOOL OF MEDICINE PROFESSOR, 医学部・医動物学教室, 教授 (20135122)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Vogt-Koyanagi-Harada disease / Cellular immunity / Cytotoxic T cell / IL-6 / Uveitis / Aqueous humor / Hybridoma |
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| Research Abstract |
Vogt-Koyanagi-Harada disease (VKH) is thought to be a systemic disorder affecting various organs containing melanocytes. Melanocyte-specific cytotoxic T lymphocytes (CTL) in VKH have been assumed to be generated and to play an important role in the manifestation of this disease.
In this study, we analyzed the blocking effects of anti-HLA class I and anti-HLA calss II antibodies against the cytotoxicity of human melanoma cell line(P-36)-specific CTL.Cytoxicities against P-36 were assayd by % specific ^<51>Cr release. The cytotoxicity against P-36 was blocked by anti-HLA calss II antibody but not by anti-HLA class I antibody. From these results P-36-specific CTL recognized the complex of melanoma antigen and HLA-class II molecules of P-36. Furthermore, we established a melanoma-specific CTL cell line(SF-1) from a patient with VKH by long term culture of peripheral blood leukocytes with P-36 under IL-2 stimulation. SF-1 showed strong cytotoxicity against P-36. Flow cytometric analysis revealed that the surface markers of SF-1 were CD4^+CD8^-HLA-DR^+CD25^+TCRalphabeta^+TCRgammadelta^-. SF-1 produced extreamly high level of IL-6 and significant levels of IL-2 and IFN-gamma. Thus, lymphokines produced by melanoma-specific CTL may play an important role in the immunopathogenesis of VKH.Additionaly, we established hybridomas of SF-1 and T cell tumor cell line(CEM). The surface markers of the hybridomas were CD2^+CD4^+CD8^-CD25^<(〕SY.+-.〔)>HLA-DR^-TCRalphabeta^<(〕SY.+-.〔)>TCRgammadelta^-CD11a^+CD28^+. Although these hybridomas adhered to P-36, it did not show cytotoxicity against P-36 and IL-2, IL-6 and IFN-gamma were not detected in the supernate of these hybridomas.
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