RESERCH ON THE RELATIONSHIP BETWEEN THE CELLULAR IMMUNITY AND CLINCAL PICTURE ON VOGT-KOYANAGI-HARADA DISEASE.

Vogt-Koyanagi-Harada病（VKH）被认为是一种影响含有黑色素细胞的各种器官的全身性疾病。黑素细胞特异性细胞毒性T淋巴细胞（CTL）被认为在VKH中产生并在该疾病的表现中起重要作用。在这项研究中，我们分析了抗hla I类和抗hla II类抗体对人类黑色素瘤细胞系（P-36）特异性CTL细胞毒性的阻断作用。通过%特异性^<51>Cr释放测定对P-36的细胞毒性。抗hlaⅱ类抗体可阻断P-36的细胞毒性，而抗hlaⅰ类抗体不能阻断P-36的细胞毒性。根据这些结果，P-36特异性CTL识别了黑色素瘤抗原和P-36的hla II类分子的复合物。此外，我们通过在IL-2刺激下用P-36长期培养VKH患者的外周血白细胞，建立了黑色素瘤特异性CTL细胞系（SF-1）。SF-1对P-36具有较强的细胞毒性。流式细胞术分析发现，SF-1的表面标记物为CD4^+CD8^- hla - dr ^+CD25^+ tcralpha ^+ tcrgamma ^-。SF-1产生极高水平的IL-6和显著水平的IL-2和ifn - γ。因此，黑色素瘤特异性CTL产生的淋巴因子可能在VKH的免疫发病机制中起重要作用。此外，我们还建立了SF-1和T细胞肿瘤细胞系（CEM）的杂交瘤。杂交瘤的表面标记物为CD2^+CD4^+CD8^- cd25 ^<(] SY.+- - [)>HLA-DR^- tcralpha ^<(] SY.+- [)> tcrgamma ^- cd11a ^+CD28^+。虽然这些杂交瘤粘附于P-36，但对P-36没有细胞毒性，并且在这些杂交瘤的上清液中未检测到IL-2、IL-6和ifn - γ。

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.