MOLECULAR IMMUNOLOGICAL STUDIES ON ITRACTABLE UVEITIS FREQUENTLY SEEN IN JAPAN.

1.白塞氏病与HLA-B51有关。本研究从Y109 YAC克隆中筛选出HLA-B和HLA-C基因，并将其亚克隆到pWE 15粘粒载体中，以寻找与该病相关的新基因。结果，Y109 YAC克隆被证明是嵌合的，并且约240 kb由染色体6区域组成，剩余的360 kb由染色体7区域组成。以pM 38粘粒克隆为探针，筛选皮肤成纤维细胞cDNA文库，分离到一个cDNA克隆。该cDNA克隆被认为位于云母和HLA-B基因之间的pM 38粘粒克隆的1.9kb EcoR Ⅰ片段上。北方杂交结果显示，在心、脑、胎盘、肺、肝和肾中有约10 kb的转录本，在骨骼肌中有约3 kb的转录本。对pM 56进行了鸟枪测序，结果显示pM 56全长39，164 bp，含有39个类似于pM 56的序列。同源性搜索最新的DNA数据库显示，5个区域高度同源的DNA报告。计算机分析这些匿名基因组序列，发现其可能存在编码序列，并可能是决定白塞病发病机制的强有力的候选基因。2.为了进行Vogt-Koyanagi-Harada病的分子免疫学研究，采用流式细胞术对眼房水淋巴细胞和脑脊液淋巴细胞进行分析。结果显示，记忆T细胞以及活化T淋巴细胞与外周血中的那些相比显著升高。因此，这些细胞在该病的免疫发病机制中起主要的免疫作用。3.总之，本研究对难治性内源性葡萄膜炎综合征的分子免疫学机制进行了进一步的探讨。这些疾病的分子遗传易感性和T细胞介导的免疫机制可能导致未来的基因诊断和基因治疗。

1.Behcet's disease is known to be associated with HLA-B51. In this study, in order to identify new gene (s) related to this disease, we screened the Y109YAC clone containing HLA-B and -C genes and subcloned into pWE15 cosmid vector. As a result, the Y109 YAC clone turned out to be chimeric and about 240 kb are consisting of the chromosome 6 region and remaining 360 kb are consisting of the chromosome 7 region. We have screened the skin fibroblast cDNA library using pM38 cosmid clone as a probe and isolated one cDNA clone. This cDNA clone was recognized to be located on the 1.9kb EcoRl fragment of the pM38 cosmid clone between the MICA and HLA-B genes. Northern hybridization showed approximately 10 kb transcript in the heart, brain, placenta, lung, liver and kidney, and 3kb transcript in the skeletal muscle. Shot-gun sequencing of the cosmid pM56 revealed to be 39,164bp in length and contained as much as 39 Alu-like sequences. Homology search for the up-to date DNA data base showed five regions highly homologous to reported DNAs. Computer analysis of these anonymous genomic sequences reveals the existence of potential coding sequences and the possibility that they are strong candidate genes for determining the pathogenesis of Behcet's disease.2.For the molecular immunological studies of Vogt-Koyanagi-Harada's disease, flowcytometric analysis were done using the aqueous humor lymphocytes and cerebrospinal fluid lymphocytes. It was revealed that memory T cells, as well as activated T lymphocytes were significantly elevated as compared with those in the peripheral blood. Therefore, these cells play main immunological roles in the immunopathogenesis of this disease.3.In conclusion, in this study, molecular immunological mechanisms of the intractable endogenous uveitis syndromes were further investigated. Molecular genetic susceptibility to these diseases, and T cell-mediated immune mechanism possibly lead to the future genetic diagnosis and gene therapy.