细胞表面β-1,4-GalT-Ⅰ在T淋巴细胞活化中的作用  

Function of β-1,4-galactosyltransferase-Ⅰ on the cell surface during the activation of T lymphocyte

作　　者：王滢[1] 吴圆圆[1] 薛琴[1] 贲志云[1] 孙晓雷[1] 胡迎青[1] 周晓荣[1] 汪晓莺[1] 

机构地区：[1]南通大学医学院微生物学与免疫学教研室,南通226001

出　　处：《现代免疫学》2011年第2期151-155,共5页Current Immunology

基　　金：国家自然科学基金资助项目(30300099)

摘　　要：本文探讨细胞表面β-1,4-半乳糖基转移酶-Ⅰ(β-1,4-galactosyltransferase-I,β-1,4-GalT-Ⅰ)在Jurkat细胞活化过程中的作用。采用植物凝集素(phytohemagglutinin,PHA)刺激Jurkat细胞,CCK-8法检测Jurkat细胞增殖;分别采用实时荧光定量PCR(RFQ-PCR)技术、Western blot技术及流式细胞术检测活化过程中的不同时间点β-1,4-GalT-Ⅰ在Jurkat细胞中的表达变化;采用免疫荧光细胞化学法和激光共聚焦技术观察Jurkat细胞表面β-1,4-GalT-Ⅰ的分布。Jurkat细胞经PHA刺激后增殖,在36 h达高峰;细胞表面β-1,4-GalT-Ⅰ的mRNA表达量、蛋白表达量以及平均荧光表达强度均较未活化状态增高;Jurkat细胞表面β-1,4-GalT-Ⅰ在未活化状态下的平均分布于细胞表面,活化后其发生重排和聚集。结果提示在T细胞活化过程中细胞表面β-1,4-GalT-Ⅰ分子的表达增高且发生了分布变化。In this report,we are attempting to explore the function of β-1,4-galactosyltransferase-Ⅰ（β-1,4-GalT-Ⅰ） on the cell surface during activation of Bunkitt＇s lymphoma Jurkat cell,in which CCK-8 was conducted to assay the cell proliferation;and real-time fluorescence quantitative PCR（RFQ-PCR）,Western blotting and flow cytometry were performed to determine the expression of β-1,4-GalT-Ⅰ at various time points after stimulation with PHA.Immunofluorescence and laser scanning co-focal microscopy techniques were employed to observe the distribution of β-1,4-GalT-Ⅰ on the surface of Jurkat T cell before and after the activation.After stimulation with PHA,the proliferation of Jurkat cell was observed.It was found that the expression of mRNA and protein and the average fluorescence value of β-1,4-GalT-Ⅰ on the cell surface were increased.During the resting status of the Jurkat cells,the β-1,4-GalT-Ⅰ was distributed on the cell surface equally.However,after activation with PHA stimulation,the membrane protein reshuffled and the β-1,4-GalT-Ⅰ became aggregated.It indicates,during the activation of Jurkat cell,the expression of β-1,4-GalT-Ⅰ has increased greatly,and the distribution of β-1,4-GalT-Ⅰ was also changed.

关 键 词：β-1 4-半乳糖基转移酶-Ⅰ 活化 T淋巴细胞 

分 类 号：R392.1[医药卫生—免疫学]