Division of Microbiology, Department of Infectious Medicine, Kurume University School of Medicine, Kurume, Japan.
Cell Microbiol. 2019 Jun;21(6):e13015. doi: 10.1111/cmi.13015. Epub 2019 Mar 5.
Epithelial cell shedding is a defence mechanism against infectious microbes that use these cells as an infection foothold and that eliminate microbes from infection foci by removing infected cells. Mycoplasma pneumoniae, a causative agent of respiratory infections, is known to adhere to and colonise the surface of ciliated airway epithelial cells; it produces a large amount of hydrogen peroxide, indicating its capability of regulating hydrogen peroxide-induced infected cell detachment. In this study, we found that M. pneumoniae reduces exogenous hydrogen peroxide-induced detachment of the infected cells from culture plates. This cell detachment occurred dependently of DNA damage-initiated, poly (ADP-ribose) polymerase 1 (PARP1)-mediated cell death, or parthanatos. In cells infected with M. pneumoniae, exogenous hydrogen peroxide failed to induce DNA damage-initiated poly (ADP-ribose) (PAR) synthesis and concomitant increased cytoplasmic membrane rupture, both of which are biochemical hallmarks of parthanatos. The impairment of PAR synthesis was attributed to a reduction in the amount of cytosolic nicotinamide adenine dinucleotide (NAD), a substrate of PARP1, caused by M. pneumoniae. On the other hand, nonadherent mutant strains of M. pneumoniae showed a lower ability to reduce cell detachment than wild-type strains, but the extent to which NAD was decreased in infected cells was comparable to that seen in the wild-type strain. We found that NAD depletion could induce PARP1-independent cell detachment pathways following stimulation with hydrogen peroxide and that M. pneumoniae could also regulate PARP1-independent cell detachment in a cytoadhesion-dependent manner. These results suggest that M. pneumoniae might regulate infected cell detachment induced by hydrogen peroxide that it produces itself, and such a mechanism may contribute to sustaining the bacterial infection.
上皮细胞脱落是一种防御机制,可防止利用这些细胞作为感染立足点的感染微生物,并通过去除感染细胞来消除感染灶中的微生物。已知肺炎支原体是呼吸道感染的病原体,它附着并定植在纤毛气道上皮细胞的表面;它产生大量的过氧化氢,表明其具有调节过氧化氢诱导的感染细胞脱落的能力。在这项研究中,我们发现肺炎支原体降低了外源性过氧化氢诱导的感染细胞从培养板上脱落。这种细胞脱落依赖于 DNA 损伤引发的多聚(ADP-核糖)聚合酶 1(PARP1)介导的细胞死亡,或 parthanatos。在感染肺炎支原体的细胞中,外源性过氧化氢未能诱导 DNA 损伤引发的多聚(ADP-核糖)(PAR)合成和伴随的细胞质膜破裂,这两者都是 parthanatos 的生化特征。PAR 合成的损伤归因于肺炎支原体引起的细胞质烟酰胺腺嘌呤二核苷酸(NAD)量减少,NAD 是 PARP1 的底物。另一方面,非黏附突变株的肺炎支原体降低细胞脱落的能力低于野生型菌株,但感染细胞中 NAD 的减少程度与野生型菌株相似。我们发现,NAD 耗竭可在刺激过氧化氢后诱导 PARP1 非依赖性细胞脱落途径,并且肺炎支原体还可以以细胞黏附依赖性方式调节 PARP1 非依赖性细胞脱落。这些结果表明,肺炎支原体可能调节自身产生的过氧化氢诱导的感染细胞脱落,这种机制可能有助于维持细菌感染。