内毒素刺激人脐静脉内皮细胞免疫相关分子的表达  

Expression of immune molecules on HUVEC stimulated by LPS

作　　者：肖宇宏[1] 白云[1] 宋敏[1] 张利永[1] 黄钢[1] 

机构地区：[1]第三军医大学基础医学部分子遗传学教研室,重庆400038

出　　处：《免疫学杂志》2006年第2期124-128,共5页Immunological Journal

基　　金：国家重点基础研究发展规划"973"资助项目(G1999054203)

摘　　要：目的了解内毒素(LPS)刺激活化人脐静脉内皮细胞(HUVEC)后,与免疫相关的膜分子和细胞因子表达及变化情况,以探讨LPS激活内皮细胞后对免疫应答可能的影响。方法胰蛋白酶法分离培养人脐静脉内皮细胞,用RT-PCR观测未刺激和用LPS刺激的内皮细胞HLA-DR,CD80、CD86、B7-H1、B7-H2、B7-H3、B7-H4、B7-DC、HVEM、TR6、LTβR、LIGHT、OX40、OX40L等免疫相关分子的mRNA表达情况;用流式细胞仪技术检测CD40、CD137I、CAM-1蛋白的表达情况;ELISA检测细胞因子IL-6I、L-8、TNF-αI、FN-γ的含量。结果①RT-PCR结果显示,未经刺激的人脐静脉内皮细胞表达B7-H2、B7-H3、TR6、LTβR,并在LPS刺激后表达上调;B7-DC、HUVEM虽呈组成性表达,但在LPS刺激后未见变化;B7-H1、B7-H4仅在内皮细胞上为诱导表达;而CD80、CD86、LIGHT、HLA-DR、OX40无论刺激与否均未表达。②流式细胞仪检测证实其细胞表面表达少量CD137、CD40及ICAM-1,LPS刺激后其表达水平明显提高;③ELISA检测发现,内皮细胞在未激活状态下就能够分泌IL-6I、L-8和TNF-α,且LPS刺激后提高其分泌量。结论人脐静脉内皮细胞组成性表达多种免疫相关分子,LPS激活内皮细胞可以显著上调其表达。人脐静脉内皮细胞由于缺少CD80、CD86的共刺激信号,不能活化初始T细胞而只能向活化及记忆性T细胞提供共刺激信号在调节特异性免疫应答的过程中,可能会起到负调控的作用。Objective To detect the expressions of immune molecules and cytokines on human umbilical vein endothelial cells （HUVEC） with or without LPS stimulation and to explore the possible influence of LPS stimulation on immune response. Methods HUVEC were obtained from healthy mature infant umbilical cord by separation and incubation. RT-PCR was used to detect the mRNA expressions of HLA-DR, CD80, CD86, B7-H1, B7-H2, B7-H3, B7-H4, B7-DC, TR6, LTβR, and LIGHT on HUVEC with or without LPS stimulation. The expressions of CD40C, D137, and ICAM-1 were assayed by FACS; the production of IL-6, IL-8, and TNF-α were detected by ELISA. Results ① HUVEC without LPS stimulation could express B7-H2, B7-H3, TR6, LTβR, B7-DC, and HVEM mRNA, except for B7-DC or HVEM. These expression levels were up-regulated by LPS. The expressions of OX40L, B7-H1, and B7-H4 mRNA were inducible. CD80, CD86, HLA-DR, and LIGHT were not detectable before and after LPS stimulation. ② HUVEC without LPS stimulation could slightly express CD40, ICAM-1, CD137, and these expression levels were obviously up-regulated by LPS. ③ HUVEC without LPS stimulation could secrete a small quantity of IL-6, IL-8, TNF-α, and LPS could up-regulated these secretions. Conclusion HUVEC could express immune molecules and these expression levels could be up-regulated by LPS stimulation. Owing to lack of CD80 and CD86, HUVEC could not activate native T cells, but could activate and regulate memory T cell and active T cell through the costimulating signal from B7 family （including B7-H1, B7-H2, B7-H3, B7-H4, and B7-DC）, thus plays a role of negative regulation in immune response.

关 键 词：人脐静脉内皮细胞 内毒素 免疫相关分子 

分 类 号：R392[医药卫生—免疫学]