PKCδ与MALT1对HIV-TB共感患者细胞免疫应答的作用

Effect of PKCδand MALT1 on Cellular Immune Response in Patients with HIV-TB Co-infection

目的分析艾滋病与结核病双重感染(HIV-TB)患者外周血单个核细胞中PKCδ与MALT1对Th17细胞水平、炎症因子水平及mRNA表达水平的影响。方法收集2021年6月-2022年8月喀什地区第一人民医院收治的HIV-TB共感患者4例,分离外周血单个核细胞(PBMC)培养后分为空白对照组和B106组[PKCδ抑制剂(B106)处理细胞]、MI-2组[MALT1抑制剂(MI-2)处理细胞]、B106+MI-2组[PKCδ抑制剂(B106)与MALT1抑制剂(MI-2)共同处理细胞]3个试验组。应用流式细胞术检测Th17细胞水平;采用ELISA法检测IL-6、IL-17、IL-23、IFN-γ、TNF-α、IL-10以及IL-22的水平及运用qPCR检测MALT1、Iκbα、P65以及IL-17、IL-23的mRNA表达水平,探讨PKCδ与MALT1对HIV-TB共感患者中细胞免疫应答的影响,并初步分析其作用机制。结果3个试验组中Th17细胞的数量均高于对照组,差异有统计学意义(P<0.05);3个试验组中IL-6、IL-17、IL-23、IFN-γ、TNF-α、IL-10以及IL-22的浓度均低于对照组,差异有统计学意义(P<0.05),但3个试验组之间比较,差异无统计学意义(P>0.05);3个试验组中IL-17、IL-23的mRNA表达水平均低于对照组,差异有统计学意义(P<0.05);B106组MALT1、P65表达低于对照组,Iκbα表达高于对照组,差异有统计学意义(P<0.05),MI-2组Iκbα表达高于对照组,P65表达低于对照组,差异有统计学意义(P<0.05);B106+MI-2组与其他2个试验组比较,差异无统计学意义(P>0.05)。结论抑制PKCδ或MALT1可拮抗HIV-TB共感患者的Th17细胞数量下降,抑制HIV-TB的炎症反应,抑制SYK/PKCδ/CARMA1-Bcl10-MALT1(CBM)复合物通路,可通过抑制NF-κB炎症通路拮抗HIV-TB的炎症反应。

Objective To analyze the effects of PKCδand MALT1 on Th17 cell level,inflammatory factor level and mRNA expression level in peripheral blood mononuclear cells for HIV-TB co-infection patients.Methods From June 2021 to August 2022,4 patients with HIV-TB co-infection were collected from the First People's Hospital of Kashgar.Peripheral blood mononuclear cells(PBMC)were isolated and cultured and divided into blank control group and B106 group[PKCδinhibitor(B106)treated cells],MI-2 group[MALT1 inhibitor(MI-2)treated cells],B106+MI-2 group[PKCδinhibitor(B106)and MALT1 inhibitor(MI-2)treated cells]three experimental groups.The level of Th17 cells was detected by flow cytometry.ELISA was used to detect the levels of IL-6,IL-17,IL-23,IFN-γ,TNF-α,IL-10 and IL-22,and qPCR was used to detect the mRNA expression levels of MALT1,Iκbα,P65,IL-17 and IL-23.The effect of PKCδand MALT1 on cellular immune response in HIV-TB co-infected patients was explored,and its mechanism was preliminarily analyzed.Results The number of Th17 cells in the three experimental groups was higher than that in the control group,and the difference was statistically significant(P<0.05).The concentrations of IL-6,IL-17,IL-23,IFN-γ,TNF-α,IL-10 and IL-22 in the three experimental groups were lower than those in the control group,and the differences were statistically significant(P<0.05),but there was no significant difference among the three experimental groups(P>0.05).The mRNA expression levels of IL-17 and IL-23 in the three experimental groups were lower than those in the control group,and the differences were statistically significant(P<0.05).The expression of MALT1 and P65 in the B106 group was lower than that in the control group,and the expression of Iκbαwas higher than that in the control group,the differences were statistically significant(P<0.05).The expression of Iκbαin the MI-2 group was higher than that in the control group,and the expression of P65 was lower than that in the control group,the differences were statistically significant(P<0.05).There was no significant difference between the B106+MI-2 group and the other two experimental groups(P>0.05).Conclusion Inhibition of PKCδor MALT1 can antagonize the decrease of Th17 cells in HIV-TB co-infected patients,inhibit the inflammatory response of HIV-TB,inhibit the SYK/PKCδ/CARMA1-Bcl10-MALT1(CBM)complex pathway,and antagonize the inflammatory response of HIV-TB by inhibiting the NF-κB inflammatory pathway.