单核细胞诱导同种异体血管内皮细胞产生干扰素诱导T细胞α型趋化因子的作用机制  

Mechanism of interferon-γ inducible T cell α chemoattractant expression during immune interaction between monocytes and allogeneic endothelial cells

作　　者：高庆贞[1] 朱彬[1] 姬凌飞[2] 王小平[1] 任万军[1] 徐何[3] 

机构地区：[1]山东大学附属济南市中心医院肾脏病-血液净化中心,250013 [2]山东省苍山县人民医院 [3]山东大学附属济南市中心医院移植科,250013

出　　处：《中华移植杂志（电子版）》2011年第1期13-17,共5页Chinese Journal of Transplantation(Electronic Edition)

基　　金：济南市科技发展计划重大项目资助课题(200705089-8)

摘　　要：目的研究外周血单核细胞与同种异体血管内皮细胞(VEC)反应产生干扰素诱导蛋白10(IP-10)和干扰素诱导的T细胞α型趋化因子(I-TAC)生成的机制。方法单独培养的VEC培养液中加入外源性TNF,用Multiplex技术检测上清液中IP-10、I-TAC浓度的变化;外周血单核细胞与同种异体VEC单独培养或共培养,检测上清液中趋化因子以及TNF浓度的变化;单核细胞和VEC共培养液中加入抗TNF抗体或核因子(NF)-κB通路阻断剂BAY11-7082干预免疫反应,观察对趋化因子生成的影响。应用实时PCR技术检测单核细胞与VEC共培养前后细胞内TNF、IP-10、I-TAC基因表达的变化。结果外源性TNF可刺激VEC产生IP-10、I-TAC;单核细胞、VEC单独培养只产生微量的IP-10、I-TAC,共培养24、48、72h后上清液中TNF以及IP-10、I-TAC浓度逐渐升高,细胞内TNF、IP-10、I-TACmRNA表达水平也显著增加;加入抗TNF抗体或BAY11-7082能消除单核细胞和VEC共培养导致的趋化因子生成。结论在同种异体单核细胞与VEC免疫反应中,单核细胞活化后产生TNF,再通过NF-κB信号通路刺激VEC产生IP-10、I-TAC等趋化因子,应用TNF抗体以及NF-κB信号通路阻断剂能减少IP-10、I-TAC等趋化因子的生成。Objective To investigate the mechanism of the production of IFN-γ inducible protein 10 （IP-10） and IFN-γ inducible T cell α chemoattractant （I-TAC） during immune interaction between monocytes and allogeneic vascular endothelial cells （VEC）. Methods Exogenous TNF was used to stimulate VEC and the amounts of IP-10 and I-TAC in the supematant were measured with Multiplex assay. A co- cultured system of monocytes and allogeneic VEC was established and the amounts of TNF, IP- 10, and I-TAC in the supernatant were detected before and after co-culture. Anti-TNF antibody or NF-kB pathway blocker, BAYll-7082, was added in the co-culture system to block their effects. The mRNA expressions of TNF, IP-10 and I-TAC were analyzed by using real-time PCR before and after co-culture. Results Multiplex assay showed an evidently increase of IP-10 and I-TAC production in the VEC supernatant after TNF stimulation. The levels of IP-IO and I-TAC were low in monocyte cultured alone and VEC supernatant but gradually elevated after 24, 48, and 72 h of co-culture. These effects were blocked with use of anti-TNF antibody or BAYll-7082. Real-time PCR analysis demonstrated up-regulation of IP-10 and I-TAC gene transcription after cell co-culture. Conclusion The interactions between monocytes and allogenic VEC can lead the production of TNF, which up-regulates the expression of IP-10 and I-TAC on VEC through NF-kBsignal transduction pathway. Anti-TNF antibody or NF-KB pathway blockade can suppress these cytokine productions.

关 键 词：单核细胞 血管内皮细胞 肿瘤坏死因子 趋化因子 排斥反应 

分 类 号：R392[医药卫生—免疫学]