背景:肺结核(PTB)是由结核分枝杆菌引起的众所周知的疾病。其发病机制的前提是逃避免疫系统和抑制免疫细胞活性。
方法:这里,免疫细胞耗竭的转录模式,炎症,在不同治疗阶段的PTB患者的外周血单核细胞(PBMC)中检查细胞活性标志物。分离PBMC,提取RNA。进行cDNA合成,然后扩增感兴趣的基因。
结果:当将0T和1T与其他治疗阶段进行比较时,T细胞耗尽标志物(PD-L1,CTLA4,CD244和LAG3)显示出不同的表达水平,提示它们作为监测结核病治疗的标志物的潜在作用。IL-2、IFN-g和TNF-a在基因水平的表达在治疗完成后恢复正常,而颗粒酶B水平在治愈阶段仍然检测不到。在固化阶段,细胞活性监测Ki67,CD69,GATA-3,CD8和CD4表达与健康对照组相当.相关分析显示与CD244表达呈显著的负相关,特别是在1T和2T之间(r=-0.94;p=0.018),和3T(r=-0.95;p=0.013)。比较0T和3T,PD-L1存在遗传相关性(r=-0.74),但无统计学意义,如CTLA4和LAG-3表达式所示。
结论:总的来说,研究结果表明,T细胞耗竭标志物特别是CD244,炎症标志物IL-2,IFN-g和TNF-a,Ki67、CD69、GATA-3、CD8和CD4等细胞活性指标是监测PTB患者治疗过程中进展的有希望的标志物。
BACKGROUND: Pulmonary tuberculosis (PTB) is a well-known disease caused by Mycobacterium tuberculosis. Its pathogenesis is premised on evasion of the immune system and dampened immune cells activity.
METHODS: Here, the transcription pattern of immune cells exhaustion, inflammatory, and cellular activity markers were examined in peripheral blood mononuclear cells (PBMCs) from PTB patients at various stages of treatment. PBMCs were isolated, and RNA extracted. cDNA synthesis was performed, then amplification of genes of interest.
RESULTS: The T cell exhaustion markers (PD-L1, CTLA4, CD244 and LAG3) showed varied levels of expressions when comparing 0 T and 1 T to the other treatment phases, suggesting their potential roles as markers for monitoring TB treatment. IL-2, IFN-g and TNF-a expression at the gene level returned to normal at completion of treatment, while granzyme B levels remained undetectable at the cured stage. At the cured stage, the cellular activity monitors Ki67, CD69, GATA-3, CD8 and CD4 expressions were comparable to the healthy controls. Correlation analysis revealed a significantly strong negative relationship with CD244 expression, particularly between 1 T and 2 T (r = -0.94; p = 0.018), and 3 T (r = -0.95; p = 0.013). Comparing 0 T and 3 T, a genitive correlation existed in PD-L1 (r = -0.74) but statistically not significant, as seen in CTLA4 and LAG-3 expressions.
CONCLUSIONS: Collectively, the findings of the study suggest that T-cells exhaustion marker particularly CD244, inflammatory markers IL-2, IFN-g and TNF-a, and cellular activity indicators such as Ki67, CD69, GATA-3, CD8 and CD4 are promising markers in monitoring the progress of PTB patients during treatment.