Lyu Xia-Li, Lin Ting-Ting, Gao Jing-Tao, Jia Hong-Yan, Zhu Chuan-Zhi, Li Zi-Hui, Dong Jing, Sun Qi, Shu Wei, Wang Sai-Sai, Pan Li-Ping, Huang Hai-Rong, Zhang Zong-De, Li Qi
Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing 101149, China.
Can J Infect Dis Med Microbiol. 2022 Apr 11;2022:2703635. doi: 10.1155/2022/2703635. eCollection 2022.
Bedaquiline (Bdq) exerts bactericidal effects against drug-susceptible and drug-resistant strains, including multidrug-resistant strains (MDR-MTBs). However, few reported investigations exist regarding Bdq effects on MDR-MTBs-infected macrophages activities and cytokine secretion. Here, Bdq bactericidal activities against MDR-MTBs and related cellular immune mechanisms were explored.
Macrophages infected with MDR-MTBs or H37Rv received Bdq treatments (4 h/8 h/24 h/48 h) at 1 × the minimum inhibitory concentration (1 × MIC), 10 × MIC and 20 × MIC. Intracellular colony-forming units (CFUs) and culture supernatant IL-12/23 p40, TNF-, IL-6, and IL-10 were determined using the Luminex® 200 system. Normally distributed continuous data (mean ± standard deviation) were analyzed using test or -test (SPSS 25.0, < 0.05 deemed statistically significant).
(1) 100% of Bdq-treated macrophages (all doses applied over 4-48 h) survived with 0% inhibition of proliferation observed. (2) Intracellular CFUs of Bdq-treated MDR-MTBs-infected macrophages decreased over 4-48 h of treatment, were lower than preadministration and control CFUs, decreased with increasing Bdq dose, and resembled H37Rv-infected group CFUs (48 h). (3) For MDR-MTBs-infected macrophages (various Bdq doses), IL-12/23 p40 levels resembled preadministration group levels and exceeded controls (4 h); TNF- levels exceeded preadministration group levels (24 h/48 h) and controls (24 h); IL-12/23 p40 and TNF- levels resembled H37Rv-infected group levels (4 h/8 h/24 h/48 h); IL-6 levels exceeded preadministration and H37Rv-infected group levels (24 h/48 h) and controls (24 h); IL-10 levels resembled preadministration and H37Rv-infected group levels (4 h/8 h/24 h/48 h) and were lower than controls (24 h/48 h); IL-12/23 p40 and IL-10 levels remained unchanged as intracellular CFUs changed, with IL-12/23 p40 levels exceeding controls (4 h) and IL-10 levels remaining lower than controls (24 h/48 h); TNF- and IL-6 levels increased as intracellular CFUs decreased (24 h/48 h) and exceed controls (24 h).
Bdq was strongly bactericidal against intracellular MDR-MTBs and H37Rv in a time-dependent, concentration-dependent manner. Bdq potentially exerted immunomodulatory effects by inducing high-level Th1 cytokine expression (IL-12/23 p40, TNF-) and low-level Th2 cytokine expression (IL-10).
贝达喹啉（Bdq）对药物敏感和耐药菌株，包括耐多药菌株（MDR-MTBs）具有杀菌作用。然而，关于Bdq对MDR-MTBs感染的巨噬细胞活性和细胞因子分泌影响的报道研究较少。在此，探讨了Bdq对MDR-MTBs的杀菌活性及相关细胞免疫机制。
用MDR-MTBs或H37Rv感染的巨噬细胞接受1×最低抑菌浓度（1×MIC）、10×MIC和20×MIC的Bdq处理（4小时/8小时/24小时/48小时）。使用Luminex® 200系统测定细胞内集落形成单位（CFUs）以及培养上清液中的IL-12/23 p40、TNF-α、IL-6和IL-10。使用t检验或方差分析（SPSS 25.0，P<0.05认为具有统计学意义）分析呈正态分布的连续数据（均值±标准差）。
（1）接受Bdq处理的巨噬细胞（4至48小时内所有剂量）100%存活，未观察到增殖抑制。（2）在4至48小时的处理过程中，Bdq处理的MDR-MTBs感染的巨噬细胞的细胞内CFUs减少，低于给药前和对照组的CFUs，随Bdq剂量增加而降低，且与H37Rv感染组的CFUs相似（48小时）。（3）对于MDR-MTBs感染的巨噬细胞（各种Bdq剂量），IL-12/23 p40水平与给药前组水平相似且超过对照组（4小时）；TNF-α水平超过给药前组水平（24小时/48小时）和对照组（24小时）；IL-12/23 p40和TNF-α水平与H37Rv感染组水平相似（4小时/8小时/24小时/48小时）；IL-6水平超过给药前和H37Rv感染组水平（24小时/48小时）以及对照组（24小时）；IL-10水平与给药前和H37Rv感染组水平相似（4小时/8小时/24小时/48小时）且低于对照组（24小时/48小时）；随着细胞内CFUs变化，IL-12/23 p40和IL-10水平保持不变，IL-12/23 p40水平超过对照组（4小时），IL-10水平低于对照组（24小时/48小时）；TNF-α和IL-6水平随着细胞内CFUs减少而增加（24小时/48小时）且超过对照组（24小时）。
Bdq对细胞内MDR-MTBs和H37Rv具有强烈的杀菌作用，呈时间依赖性和浓度依赖性。Bdq可能通过诱导高水平的Th1细胞因子表达（IL-12/23 p40、TNF-α）和低水平的Th2细胞因子表达（IL-10）发挥免疫调节作用。
