摘要
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。
Objective:To study the synthesis, assembly, transportation and expression of cell surface and soluble HLA B27 molecules in human lymphoblastoid cell line C1R.Methods:B27 and soluble B27 (sB27) cDNA were cloned into eukaryotic expression vector RSV.5neo and transfected into C1R cells through electroporation. Monoclonal antibodies were used in experiments such as immunoprecipitation, pulse chase and endoglycosidase H digestion to study the cell biology of B27 and sB27 molecules and their association with molecular cheparones Calnexine and BiP.Results:The synthesis, assembly, transpotation and expression of sB27 molecule were completed through the same pathway inside C1R cell line as cell surface B27 molecules although only part of the sB27 molecules finished the whole process. Most of the sB27 molecules were retained inside the endoplasmic reticulum. Conclusion:As a product of alternative splicing of B27 molecules, sB27 molecule which had been released to the supernatant had similar cell biology to its cell surface counterpart. It is speculated that most of the sB27 molecules retained inside endoplasmic reticulum would be degraded and presented to CD4 + T cells. through class Ⅱ molecules. This hypothesis explained the recent findings that B27 related spondyloarthropathies had HLA class Ⅱ antigen association clues such as CD4 + T cell activation.
出处
《军医进修学院学报》
CAS
2000年第3期161-164,共4页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金资助项目!(39770704)
国家教委留学回国人员科研启动基金资助!(教外司留 436号)
