Ilbeigi Parisa, Sadraei Javid, Ghaffarifar Fatemeh, Siadat Seyed Davar, Mirjalali Hamed
Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
Mol Biol Rep. 2025 Jul 16;52(1):728. doi: 10.1007/s11033-025-10823-6.
The control of T. gondii in macrophages is mediated by the P2X7 receptor. The P2X7 receptor plays a crucial role in defense against Mycobacterium tuberculosis (M.tb), as well. Current study aimed to investigate the immunity induced by T. gondii soluble total antigen (STAg) against M.tb via P2X7R in human monocyte cell line.
The STAg was prepared from 1.6 × 10 of fresh T. gondii tachyzoites, RH strain. The siRNA targeting P2X7R was employed to knockdown P2X7R in M0 THP-1 macrophages. Prior to M.tb infection, the cells were treated by 80 µg/mL of STAg for 4 h. The cells were infected by M.tb with multiplicity of infection (MOI) 5 for 3 h. The expression of the ATG5, ATG7, ATG16, P2X7R genes and the number of up taken M.tb were investigated by qReal-time PCR and absolute qReal-time PCR, respectively.
The qReal-time PCR analysis showed overexpression of the ATG5 gene in P2X7R siRNA, scramble siRNA, and STAg/siRNA and ATG7 in THP-1 cell line treated by STAg groups, while they were suppressed in STAg/siRNA/ M.tb and STAg/ M.tb groups. The expression of ATG16 was significantly overexpressed in STAg group, while it was suppressed in treated wells by STAg/siRNA, STAg/siRNA/M.tb, STAg/M.tb, and M.tb. The number of entered M.tb in STAg/M.tb was significantly lower than M.tb group. The number of M.tb in P2X7R-knockdown THP-1 cells was significantly higher than scramble siRNA and STAg. In addition, the number of the bacteria in STAg/siRNA was significantly lower than P2X7R siRNA group.
Our study showed that sensing macrophages by T. gondii STAg may induce levels of immunity against M.tb. In addition, our findings support available data, indicating potential role of P2X7R in macrophage against T. gondii and M.tb.
巨噬细胞中弓形虫的控制由P2X7受体介导。P2X7受体在抗结核分枝杆菌（M.tb）防御中也起着关键作用。当前研究旨在调查弓形虫可溶性总抗原（STAg）通过人单核细胞系中的P2X7R诱导的针对M.tb的免疫力。
从1.6×10个新鲜的弓形虫速殖子RH株制备STAg。使用靶向P2X7R的siRNA敲低M0 THP-1巨噬细胞中的P2X7R。在M.tb感染前，细胞用80μg/mL的STAg处理4小时。细胞以感染复数（MOI）5感染M.tb 3小时。分别通过q实时PCR和绝对q实时PCR研究ATG5、ATG7、ATG16、P2X7R基因的表达以及摄取的M.tb数量。
q实时PCR分析显示，在P2X7R siRNA、乱序siRNA和STAg/siRNA处理的THP-1细胞系中，ATG5基因过表达，在STAg处理组中ATG7过表达，而在STAg/siRNA/M.tb和STAg/M.tb组中它们受到抑制。ATG16的表达在STAg组中显著过表达，而在STAg/siRNA、STAg/siRNA/M.tb、STAg/M.tb和M.tb处理的孔中受到抑制。STAg/M.tb中进入的M.tb数量显著低于M.tb组。P2X7R敲低的THP-1细胞中的M.tb数量显著高于乱序siRNA和STAg组。此外，STAg/siRNA中的细菌数量显著低于P2X7R siRNA组。
我们的研究表明，弓形虫STAg感知巨噬细胞可能诱导针对M.tb的免疫水平。此外，我们的发现支持现有数据，表明P2X7R在巨噬细胞对抗弓形虫和M.tb中的潜在作用。
