Krämer S, Crauwels P, Bohn R, Radzimski C, Szaszák M, Klinger M, Rupp J, van Zandbergen G
Division of Immunology, Paul Ehrlich Institute, Federal Institute for Vaccines and Biomedicines, Langen, Germany.
Institute of Medical Microbiology and Hygiene, University of Luebeck, Luebeck, Germany.
Infect Immun. 2015 Jul;83(7):2651-60. doi: 10.1128/IAI.03083-14. Epub 2015 Apr 20.
Chlamydia pneumoniae is a Gram-negative bacterium that causes acute or chronic respiratory infections. As obligate intracellular pathogens, chlamydiae efficiently manipulate host cell processes to ensure their intracellular development. Here we focused on the interaction of chlamydiae with the host cell transcription factor activator protein 1 (AP-1) and its consequence on chlamydial development. During Chlamydia pneumoniae infection, the expression and activity of AP-1 family proteins c-Jun, c-Fos, and ATF-2 were regulated in a time- and dose-dependent manner. We observed that the c-Jun protein and its phosphorylation level significantly increased during C. pneumoniae development. Small interfering RNA knockdown of the c-Jun protein in HEp-2 cells reduced the chlamydial load, resulting in smaller inclusions and significantly lower chlamydial recovery. Furthermore, inhibition of the c-Jun-containing AP-1 complexes using tanshinone IIA changed the replicative infection phenotype into a persistent one. Tanshinone IIA-dependent persistence was characterized by smaller, aberrant inclusions, a strong decrease in the chlamydial load, and significantly reduced chlamydial recovery, as well as by the reversibility of the reduced recovery after the removal of tanshinone IIA. Interestingly, not only was tanshinone IIA treatment accompanied by a significant decrease of ATP levels, but fluorescence live cell imaging analysis by two-photon microscopy revealed that tanshinone IIA treatment also resulted in a decreased fluorescence lifetime of protein-bound NAD(P)H inside the chlamydial inclusion, indicating that chlamydial reticulate bodies have decreased metabolic activity. In all, these data demonstrate that the AP-1 transcription factor is involved in C. pneumoniae development, with tanshinone IIA treatment resulting in persistence.
肺炎衣原体是一种革兰氏阴性细菌,可引起急性或慢性呼吸道感染。作为专性胞内病原体,衣原体有效地操纵宿主细胞过程以确保其在细胞内的发育。在此,我们聚焦于衣原体与宿主细胞转录因子激活蛋白1(AP-1)的相互作用及其对衣原体发育的影响。在肺炎衣原体感染期间,AP-1家族蛋白c-Jun、c-Fos和ATF-2的表达和活性呈时间和剂量依赖性调节。我们观察到,在肺炎衣原体发育过程中,c-Jun蛋白及其磷酸化水平显著增加。在HEp-2细胞中通过小干扰RNA敲低c-Jun蛋白可降低衣原体载量,导致包涵体变小且衣原体回收率显著降低。此外,使用丹参酮IIA抑制含c-Jun的AP-1复合物可将复制性感染表型转变为持续性感染表型。丹参酮IIA依赖性持续性感染的特征为包涵体更小、异常,衣原体载量大幅下降,衣原体回收率显著降低,以及在去除丹参酮IIA后回收率降低的可逆性。有趣的是,丹参酮IIA处理不仅伴随着ATP水平的显著降低,而且通过双光子显微镜进行的荧光活细胞成像分析显示,丹参酮IIA处理还导致衣原体包涵体内与蛋白结合的NAD(P)H荧光寿命缩短,表明衣原体网状体的代谢活性降低。总之,这些数据表明AP-1转录因子参与肺炎衣原体的发育,丹参酮IIA处理导致持续性感染。