艾滋病免疫重建不全患者Toll样受体信号转导通路改变及免疫2号方干预的影响

目的:采用基因芯片技术,从Toll样受体信号转导通路的角度,探讨艾滋病免疫重建不全患者的免疫失调机制及中药免疫2号方的干预作用。方法:采集4例艾滋病免疫重建不全患者疗前以及联合中药免疫2号方干预6个月后的外周血单个核细胞(PBMC),另以4例HIV抗体阴性健康献血员做对照,抽提mRNA,通过逆转录后以TLRs信号通路实时定量PCR芯片进行杂交,采用ΔΔCt方法计算RNA表达量,寻找TLR通路上显著表达的差异基因。结果:筛选出与艾滋病免疫重建不全有关的基因显著上调≥2倍的有5个(干扰素-γ、IL-6、IL-8等),下调≥2倍的基因有4个(TLR9、NF-κB等),通路上游以基因下调改变为主(TLR9、TOLLIP、TRIF等),下游则多为上调改变(AP-1、IL-6、IL-8)。联合免疫2号方干预后,相关基因显著上调≥2倍的有7个(IL-2、IL-10、TLR1等),无显著下调基因,对通路上游基因的上调作用最为显著(如TLR1、CD14、TRAF6)。结论:HAART免疫重建不全的艾滋病患者细胞内TLR9较正常人表达低下,IRAK1、NF-κB表达下调,提示TLR9介导的信号转导系统异常是艾滋病患者免疫重建不全发生的可能机制之一;免疫2号方调节机体免疫功能的机制可能主要通过上调TLR1和CD14表达,进而引起下游一系列通路信号分子传递而实现。

Changes on Toll-like receptor signalling pathway of AIDS patients with incomplete immune reconstitution and the influence of Immune No.2 by gene chip technology

Objective: From the Toll like receptor signaling pathway,to discuss the mechanism of incomplete immune reconstruction of AIDS patients and the effect of Immune No.2 by TLRs signaling pathway real-time PCR microarrays.Methods: PBMCs were isolated from 4 cases of AIDS patients failure to immune reconstitution before and after treatment with Immune No.2 and 4 cases of HIV negative healthy donors.mRNA were extracted,and first strand cDNA were synthesis,then real time PCR microarrays were performed.ΔΔCt method was used to calculate the RNA expression.Results: There were 5 genes involved in the complete immune reconstruction significantly raised more than 2 times,and 4 genes decreased.Genetic changes in upstream were mainly down regulated,while in downstream were mainly up regulated.After treatment of Immune No.2,there were 7 related genes significantly raised more than 2 times,and no genes significant decreased.Genetic changes in upstream were mainly up regulated.Conclusion: TLR9 expressed lowly in AIDS patients with incomplete immune reconstitution than in healthy person,and also IRAK1、NF-κB,suggesting that abnormal signal pathway mediated by TLR9 maybe one of the mechanisms of incomplete immune reconstitution of AIDS.The effect of Immune No.2 may be realized by up regulating TLR1 and CD14 expression,resulting changes of signal molecules in downstream.