卡介苗(BCG)通过诱导IL-12产生和受体表达促进人NK细胞功能  

Bacillus Calmette-Guérin enhances the function of human nature killer cells by inducing IL-12 production and IL-12R expression

作　　者：周晖[1,2] 范艳莹[2] 吴长有[2] 

机构地区：[1]广东省医学科学院广东省人民医院检验科,广东广州510080 [2]中山大学中山医学院免疫学教研室热带病防治研究教育部重点实验室,广东广州510080

出　　处：《细胞与分子免疫学杂志》2009年第11期976-979,983,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30671898);教育部博士学科点专项科研基金资助项目(20070558213);广东省医学科研基金资助项目(A2009161);广东省自然科学基金博士启动项目(8451008901000131)

摘　　要：目的:研究卡介苗(BCG)对人自然杀伤细胞(NK)功能的作用及其机制。方法:分离抗结核抗体阴性志愿者外周血PBMC、纯化NK细胞,分别与BCG、IL-12、BCG+IL-12、BCG+抗IL-12Rβ1 mAb(2B10)培养。利用ELISA方法检测培养上清液IFN-γ、IL-12p40含量;利用ELISpot方法检测IFN-γ、颗粒酶B产生细胞的频率;利用四甲基偶氮唑盐(MTT)比色法测定杀伤功能。利用流式细胞术检测NK细胞IL-12Rβ1的表达。结果:BCG呈剂量依赖的方式诱导PBMC产生IFN-γ。在BCG刺激条件下,PBMC颗粒酶B分泌细胞数明显高于不加任何刺激剂组(P<0.05)。BCG增强PBMC杀伤活性。BCG不能诱导纯化NK细胞产生IFN-γ,但与IL-12同时刺激则表现出协同作用。纯化NK细胞经BCG刺激后杀伤活性与未刺激相比差异无统计学意义。BCG呈剂量依赖方式诱导PBMC产生IL-12、并促进NK细胞不同亚群表达IL-12Rβ1。2B10抗体抑制BCG对PBMC产生IFN-γ和分泌颗粒酶B的诱导作用。结论:BCG间接地促进NK细胞的生物学活性,其部分机制是通过诱导单核细胞产生内源性IL-12、并上调NK细胞表达IL-12R。AIM： To study the effect and mechanism of BCG on human nature killer cells. METHODS： PBMC or purified NK cells were isolated from normal human peripheral blood with negative anti-Mycobacterium tuberculosis antibody and cultured with BCG, IL-12, BCG plus IL-12 and BCG plus anti-lL-12Rβ1 mAb （2B10）, respectively. The levels of IFN-γ and IL-12p40 in the culture supernatants were measured by ELISA. The frequency of IFN-γ and granzyme B producing cells were analyzed by ELISpot. The cytolytic activity was detected by MTT- reduction assay. The surface expression of IL-12Rβ1 on NK cells was detected by flow cytometry. RESULTS： BCG significantly induced IFN-γ production by PBMC in a dose-dependent manner. When PBMC was stimulated with BCG, the frequency of granzyme B producing cells was higher than that in unstimulated PB-MC （ P 〈 0.05）. BCG enhanced the cytotoxic activity of PB- MC. BCG alone didn＇t induce IFN-γ production by purified NK cells, but it can augment IL-12-induced IFN-γ production by purified NK cells. The cytotoxic activities of BCG-stimulated and unstimulated purified NK cells were not significantly different （P〉0.05）. BCG induced IL-12 production by PB-MC in a dose-dependent manner and enhanced IL-12Rβ1 expression on different subsets of NK cells. Blocking the effect of IL-12 by anti-lL-12Rβ1 mAb （2B10） inhibited BCG- induced IFN-γ production and granzyme B releasing by PB- MC. CONCLUSION： BCG can indirectly promote biologic activity of NK cells and the production of endogenous IL-12 combined with up-regulation IL-12Rβ1 expression on the surface of NK cells is a part of the mechanisms of IL-12 on human NK cells.

关 键 词：卡介苗 自然杀伤细胞 外周血单个核细胞 细胞因子 细胞毒性 

分 类 号：R392.12[医药卫生—免疫学]