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目的 :了解两型HLA B2 7分子在人细胞系内的合成、组装及转运过程。方法 :克隆到真核表达载体RSV 5neo的膜表面型与可溶型B2 7(sB2 7)cDNA以电转法转入人B淋巴母细胞突变细胞系C1R ,并使其稳定表达。以免疫沉淀、脉冲追踪及内切糖苷酶H消化实验观察两型B2 7分子在C1R细胞系内的细胞生物学特点。结果 :sB2 7分子与膜表面型B2 7分子一样 ,约需 30min完成细胞内的修饰过程 ,均与Calnexin及BiP这两种分子伴侣相关等 ,但与膜表面型B2 7分子相比 ,仅部分sB2 7分子被转运到细胞表面 ,相当一部分sB2 7分子 4h后仍滞留在内质网腔内。结论 :sB2 7分子在人C1R细胞内的合成、组装与转运过程与膜表面型B2 7分子相似 ,滞留在内质网腔的sB2 7分子的酶解片段有可能通过Ⅱ类分子而递呈给CD4+T细胞 ,这一现象将为研究HLA Ⅱ类分子与脊柱关节病的相关提供新线索 ,并为今后探讨B2 7及环境因素与脊柱关节病的相关、寻找早期诊断与早期治疗措施提供理论依据
OBJECTIVE: To understand the synthesis, assembly and transport of two types of HLA B2 7 molecules in human cell lines. METHODS: The membrane-surface type and soluble B2 7 (sB2 7) cDNA cloned into eukaryotic expression vector RSV 5neo was electrotransformed into human B lymphoblastoid cell line C1R and stably expressed. Immunoprecipitation, pulse tracing and endoglycosidase H digestion were used to observe the cell biology of two B2 7 molecules in C1R cell line. Results: As with the B2 7 molecule on the membrane surface, the sB2 7 molecule takes about 30 minutes to complete the intracellular modification process and is related to the two chaperones Calnexin and BiP. However, compared with the B2 7 molecule on the membrane surface, only part of the sB2 7 molecules were transported to the cell surface, a considerable part of sB2 7 molecules remain in the endoplasmic reticulum after 4h. CONCLUSIONS: The synthesis, assembly and transport of sB2 7 molecule in human C1R cells are similar to the membrane-surface type B2 7 molecule. The digested fragment of sB2 7 residing in the endoplasmic reticulum cavity may be presented to class II molecules CD4 + T cells, this phenomenon will provide new clues for the study of the correlation between HLA class II molecules and spondyloarthropathy, and provide theoretical basis for the future study on the relationship between B2 7 and environmental factors and spondyloarthropathies, looking for early diagnosis and early treatment measures in accordance with