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

Vogt-Koyanagi-Harada病(VKH)被认为是一种全身性疾病，影响含有黑素细胞的各种器官。黑素细胞特异性细胞毒性T淋巴细胞(CTL)在VKH的发生和发展中起重要作用。本研究分析了抗HLAI类和抗HLACalss II抗体对人黑色素瘤细胞系(P-36)特异性CTL的阻断作用。对P-36的杀伤作用可被抗HLACalss II抗体阻断，但不能被抗HLAI类抗体阻断。根据这些结果，P-36特异性CTL识别黑色素瘤抗原和P-36的HLAII类分子的复合体。此外，我们建立了黑色素瘤特异性CTL细胞系(SF-1)，来自VKH患者，通过在IL-2刺激下用P-36长期培养外周血白细胞。SF-1对P-36有较强的细胞毒作用。流式细胞仪分析表明，SF-1的表面标志为CD4^+CD8^-HLA-DR^+CD25^+TCRalphabeta^+TCRgammadelta^-.SF-1产生极高水平的IL-6和显著水平的IL-2和干扰素-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与T细胞肿瘤细胞株(CEM)的杂交瘤。杂交瘤细胞的表面标记为CD2^+CD4^+CD8^-CD25^&lt；(〕SY.+-.〔)&gt；HLA-DR^-TCRalphabeta^&lt；(〕SY.+-.〔)&gt；TCRgammadelta^-CD11a^+CD28^+.虽然这些杂交瘤细胞与P-36黏附，但对P-36无细胞毒作用，并且在杂交瘤细胞培养上清液中未检测到IL-2、IL-6和干扰素-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.