Ipr1对巨噬细胞抗结核分枝杆菌感染免疫相关基因表达的影响

Immune response genes expressing of Ipr1 expressed macrophage anti-M.tuberculosis

目的应用基因芯片技术检测Ipr1的表达对巨噬细胞感染分枝杆菌H37Ra后参与免疫应答的相关基因的表达差异。方法实验组与对照组细胞分别感染分枝杆菌H37Ra,基因芯片检测实验组和对照组细胞感染H37Ra 96h后免疫相关基因表达差异。定量PCR反应检测芯片结果中上调的3个基因的表达差异用以验证芯片结果的可靠性。结果基因芯片检测结果显示Ipr1基因的表达上调了11个固有免疫机制中相关基因表达,其中与MΦ抗Mtb感染关系密切的基因有:TLR2、TLR4I、rak1、Traf6、Ifngr1、Tnfrsf1a,而参与调节适应性免疫应答的相关基因如:IL-1,IL-12无明显表达差异。结论 Ipr1增强MΦ抗Mtb感染的机制可能主要是通过上调MΦ抗感染固有免疫机制的有关基因的表达从而发挥抗菌作用。本实验为进一步研究Ipr1促进MΦ的活化及杀伤胞内吞噬的Mtb的作用机制奠定了基础。

To study the effect and mechanisms of Iprl gene expressing in macrophages infected with Mycobacteriurn tuberculosis H37Ra use cDNA microarry. The experimental group RAW264. 7 cells were transfected with pEGFP-Iprl and the stable Iprl gene expressed RAW264.7 cells were selected by G418. The control group RAW264.7 ceils were transfeeted with pEGFP-Cland selected by G418 . Experimental group and control group cells were infected by Mycobacterium tuberculosis H37Ra. After 96h infection, the total RNA were isolated from the two groups. Then using a cDNA microarry（Mouse Innate and Adaptive Immune Responses Microarray 113 genes） to detect the differences of experimental group and control group in gene expression related to immune response. Real-time quantitative PCR test was used to verify the reliability of the chip re- suits. The cDNA microarry result display that Iprl gene expression up regulated 11 genes for macrophage anti Mtb innate immunity involved TLRs signaling pathway： TLR2 and TLR4, Irakl, Traf6 ; as well as interferon-related genes： Ifngrl （IFN-y R1） and tumor necrosis factor-related genes： Tnfrsfla （TNFR1）,et al. And there was no significant difference about the mo- lecular expression involved in the regulation of the adaptive immune response such as： IL 1, IL12. The results of real-time PCR reaction TLR2 and TLR4 and Ifngrl indicated that the real-time PCR results consistent with the trend of chip results. The results of gene chip analysis point that the Iprl gene expression maybe enhance macrophage anti-Mtb infection by the mecha nisms of increasing the innate immunity gene expression, in particular TLR2/TLR4 and its signal transduction molecules as well as the IFN -TR1, TNFR1 expression, and promote macrophage activation and intracellular anti-Mtb effect. This constitutes the basis for further study of the mechanisms of Iprl gene in host innate immunity against tuberculosis infection.