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Biological Significance of Nuclear Antigens' Expression on Cell Surface

细胞表面核抗原表达的生物学意义

基本信息

批准号：
01570565
负责人：
FURUKAWA Fukumi
金额：
$ 1.34万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

1) In order to understand the potential mechanisms of ultraviolet light B (UVB) on photosensitivity in lupus erythematosus ( LE), we designed immunopathologic in vitro and in vivo experiments to evaluate the effects of UV on the binding of anti-extractable nuclear antigent (ENA) antibodies to the surface of human keratinocytes. Short-term 2% paraformaldehyde fixation of suspensions of cultured human keratinocytes previously incubated with monospecific antiserum probes enabled the detection of ENA expression on the cell surface by flow cytometry analysis. In vitro FACS analysis revealed that ENA augmentation on the keratinocyte cell surface was UVB dose dependent, glycosilation dependent, and cell cycle independent. In vivo augmentation on the keratinocyte surface was also demonstrated in suction blister epidermal roof.2) Among PG reagents, PGJ2 could augment the binding of anti-ENA antibodies to cultured keratinocytes in serum-free medium. However, anti-Smantibody's binding was not ind … More uced by PGJ_2. Enhanced bindings of anti-RNP, anti-SS-A/Ro antibodies were demonstrated by the standard immunofluorescence technique and the FACS analysis using post-fixation methods. These binding were blocked by cyclohexamide.These results suggest that the inducers of heat shock protein or stress protein are triggers of development of skin manifestation of cutaneous LE.3) As a next step, the cytotoxic mechanisms of cutaneous LE were investigated. Monocytes from SLE or subacute cutaneous LE were co-cultured with keratinocytes treated with PGJ_2 or cells irradiated with UVB. Irrespective of the presence or the absence of complement source, the significant number of cultured keratinocytes was damaged under the influence of antiserum probes.These results indicate that the cytotoxic mechanisms such as antibody dependent cell-mediated cytotoxicity (ADCC) are involved in the pathogenesis of cutaneous LE.4) Expression of nuclear antigens on the cells of blood vessels was also investigated. UVB could induce anti-RNP antibody binding of cryostat sections of skin specimens. The bindings were observed on the endothelial cells of dermis. The specificity of this positive findings is now under careful in stigation. Less

1)为了解紫外线B（UVB）对红斑狼疮（LE）光敏性的影响，我们设计了体内外免疫病理学实验，观察UV对抗可提取核抗原（ENA）抗体与人角质形成细胞表面结合的影响。短期的2%多聚甲醛固定的悬浮液培养的人角质形成细胞先前孵育的单特异性抗血清探针，使ENA表达的细胞表面上的流式细胞仪分析检测。体外流式细胞仪分析表明，ENA对角质形成细胞表面的扩增是UVB剂量依赖性的，糖基化依赖性的，和细胞周期无关。2）在PG试剂中，PGJ 2可以增强抗ENA抗体与无血清培养的角质形成细胞的结合。然而，抗Smantibody的结合并不明显。 ...更多信息由PGJ_2执行。通过标准免疫荧光技术和使用固定后方法的FACS分析证实抗RNP、抗SS-A/Ro抗体的增强结合。这些结果表明热休克蛋白或应激蛋白的诱导剂是皮肤LE皮肤表现发展的触发剂。3）作为下一步，研究皮肤LE的细胞毒性机制。将SLE或亚急性皮肤LE的单核细胞与PGJ_2处理的角质形成细胞或UVB照射的细胞共培养。无论有无补体来源，在抗血清探针的作用下，大量培养的角质形成细胞受到损伤，这些结果表明抗体依赖的细胞介导的细胞毒性（ADCC）等细胞毒性机制参与了皮肤LE的发病机制。4）还研究了血管细胞核抗原的表达。UVB可诱导皮肤标本冰冻切片中的抗RNP抗体结合。在真皮内皮细胞上观察到结合。这一阳性结果的特异性目前正在仔细研究中。少