重组hIFN—α-2b—BCG对人外周血单个核细胞的TLR4通路的免疫活化作用  

Toll-like receptor 4 expression of human peripheral blood mononulcar cells treated by recombinant hIFN-α-2b-BCG and its role of immune activation

作　　者：杨雄[1] 孙二琳[1] 史启铎[1] 韩瑞发[1] 

机构地区：[1]天津医科大学第二医院天津市泌尿外科研究所天津市泌尿外科基础医学重点实验室,300211

出　　处：《中华医学杂志》2012年第20期1419-1423,共5页National Medical Journal of China

基　　金：国家科技部“十一五”重大专项（2009ZX091034599）;天津市科技支撑计划重大项目（07ZCKFSH03200）;天津卫生局重点攻关项目（11KG147）

摘　　要：目的了解重组卡介苗（BCG）对人外周血单个核细胞（hPBMC）T0u样受体4（TLR4）的表达调节及其介导免疫细胞活化效应的机制。方法比较重组BCG和野生BCG对hPBMC的TLR4表达的调节；应用TLR4功能阻断抗体对hPBMC的TLR4信号通路进行干预，然后用重组人干扰素（hIFN）-α-2b—BCG（重组BCG）、野生BCG、hIFN—α-2b和磷酸盐缓冲液（PBS）（空白对照）对TLR4信号通路阻断和未阻断的hPBMC进行刺激，并运用ELISA方法比较阻断组和未阻断组人肿瘤坏死因子（hTNF）a、人白细胞介素（hIL）12、hIFN-1表达量的变化。结果重组BCG和野生BCG对hPBMC的TLR4表达较空白对照组均有明显正性调节作用（均P〈0．05）。TLR4功能未阻断时，hIFN-γ和hTNF-α的表达量在不同刺激组均为重组BCG组〉野生BCG组〉hIFN—α-2b组〉空白对照组，而hIL-12的表达量在不同刺激组为hIFN-α-2b组〉空白对照组〉重组BCG组〉野生BCG组（均P〈0．05）。TLR4功能阻断后48h，重组BCG组、野生BCG组、hIFN—α-2b组和空白对照组的hIFN-γ的表达量（27．3±1．2、20．6±0．9、20．3±0．8、18．4±0．7）均明显低于未阻断组（84．6±1．3、34．0±1．0、24．9±0．9、22．9±0．7）（均P〈0．05）；hTNF-α的表达量（1431±28、1032±21、104±6、109±4）均明显低于未阻断组（1553±28、1065±31、343±6、299±4）（均P〈0．05）；hIL-12的表达量（0．646±0．005、0．592±0.015、0．638±0．008、0．595±0．019）均明显低于未阻断组（1．120±0．012、0．946±0．015、1．254±0．011、1．112±0．024）（均P〈0．05）。结论重组BCG可通过TLR4信号通路对hPBMC分泌Th1细胞因子的表达量进行调节。Objective To examine the toll-like receptor 4 （TLR4） expression of human peripheral blood mononuclear cells （hPBMC） treated with recombinant bacillus Calmette-Gu6rin （rBCG）and its role of immune activation. Methods hPBMC was treated with recombinant human interferon （hIFN）-α-2b-BCG （rBCG）or wild BCG（wBCG） in vitro and the TLR4 expression detected by flow cytometry. The TLR4 functional blocking antibodies were applied for intervening the TLR4 signaling pathway of hPBMC. Then rBCG, wBCG, hIFN-α-2b and phosphate-buffered solution （PBS） were used to stimulate the hPBMC of blocking and non-blocking groups. The changes of human tumor necrosis factor-alpha （ hTNF-α）, human interleukin-12 （hlL-12） and hIFN--γ between the blocking and non-blocking groups by ELISA. Results The expression of TLR4 in hPBMC treated with rBCG or wBCG were stronger than that treat with PBS （all P 〈 0. 05）. In TLR4 non-blocking groups the expressions of hTNF-α and hIFN-γ were rBCG group 〉 wBCG group 〉 hIFN-ct-2b group 〉 PBS group, the expressions of hIL-12 was hIFN-α-2b group 〉 PBS group 〉 rBCG group 〉 wBCG group（ all P 〈 0. 05）. Application of TLR4 functional blocking antibodies to intervene hPBMC 48 h, comparing the changes in rBCG group, wBCG group, hIFN-α-2b group and PBS group, the expressions of hlFN-α/in TLR4 blocking groups （27. 3 ± 1.2,20. 6 ± 0. 9,20. 3 ± 0. 8,18.4 ± 0. 7 ） were significantly inhibited than those in non-blocking groups （84. 6 ± 1.3,34. 0 ± 1.0,24. 9 ＋ 0.9,22. 9 ±0. 7 ） （ all P 〈 0. 05 ）. The expressions of hTNF-ot in TLR4 blocking groups （ 1431 ± 28,1032 ± 21,104 ±6,109 ± 4） were significantly inhibited than those in non-blocking groups （1553 ± 28,1065 ± 31,343 ± 6,299 ± 4 ） （ all P 〈 0. 05 ）. The expressions of hlL-12 in TLR4 blocking groups （ 0. 646 ± 0. 005,0. 592 ± 0. 015, 0. 638± 0. 008,0. 595 ± 0. 019 ） were significantly inhibited than those in non-blocking groups （ 1. 120 ± 0.012,0.946±0

关 键 词：卡介苗 干扰素Α-2B Toll样受体4 细胞因子类 外周血单个核细胞 

分 类 号：R392[医药卫生—免疫学]