HLA-B27分子与分子伴侣的相关性研究

The assembly and transportation of HLA B27 molecule and its relationship with molecular chaperone calnexin and BiP

目的观察ＨＬＡＢ２７分子与Ｃａｌｎｅｘｉｎ及ＢｉＰ两种分子伴侣的相关，了解ＨＬＡ分子在人体细胞内的合成、组装及转运过程。方法将人ＨＬＡＢ２７基因转入人Ｂ淋巴母细胞系Ｃ１Ｒ，以单克隆抗体、免疫沉淀、脉冲追踪及内切糖苷酶Ｈ消化实验判断Ｂ２７分子在细胞内的成熟过程，及Ｂ２７分子与两种分子伴侣之间的相关性。结果Ｃ１Ｒ细胞在加入培养基３０分钟时已开始将新合成的Ｂ２７分子从内质网转运到高尔基体，２小时后所有新合成的Ｂ２７分子完成在高尔基体内的修饰过程，并被转运到细胞表面。结论Ｃａｌｎｅｘｉｎ及ＢｉＰ均参与Ｂ２７分子在细胞内的组装过程，主要与新生态的分子结合。本研究为今后探讨Ｂ２７与强直性脊柱炎的相关。

Objective To investigate the cell biology of HLA B27 molecule and its association with molecular chaperones calnexin and BiP.Methods Human HLA B27 gene cDNA was transfected into mutant human B cell line,C1R.Using monoclonal antibodies,immunoprecipitation,pulse chase and endoglycosidase H digestion experiment,the expression,assembly and transportation of B27 molecules were studied.The associations of B27 molecule with calnexin and BiP were also studied.Results The results showed that the C1R cells start to synthesize B27 molecules right after the addition of culture medium.About 30 minutes later,some nascent B27 heavy chains already completed their glycosylation in the Golgi apparatus and became endo H resistant.The whole maturation process took about 2 hours.Conclusion The association of calnexin and BiP with B27 was mainly with nascent B27 molecules.The understanding of the cell biology of B27 in human cells plays an important role in future studies such as how bacteria infection influences the assembly and expression of surface and soluble B27 molecules.