Institute of Veterinary Pathology, University of Zurich, Switzerland.
BMC Microbiol. 2010 Jul 27;10:201. doi: 10.1186/1471-2180-10-201.
Chlamydiae induce persistent infections, which have been associated with a wide range of chronic diseases in humans and animals. Mixed infections with Chlamydia and porcine epidemic diarrhea virus (PEDV) may result in generation of persistent chlamydial infections. To test this hypothesis, an in vitro model of dual infection with cell culture-adapted PEDV and Chlamydia abortus or Chlamydia pecorum in Vero cells was established.
Infected cultures were investigated by immunofluorescence (IF), transmission electron microscopy (TEM) and re-infection experiments. By IF, Chlamydia-infected cells showed normal inclusions after 39 hpi. Dual infections with Chlamydia abortus revealed a heterogenous mix of inclusion types including small inclusions consisting of aberrant bodies (ABs), medium-sized inclusions consisting of ABs and reticulate bodies and normal inclusions. Only aberrant inclusions were observable in dual infection experiments with Chlamydia pecorum and PEDV. TEM examinations of mixed infections with Chlamydia abortus and Chlamydia pecorum revealed aberrant chlamydial inclusions containing reticulate-like, pleomorphic ABs, which were up to 2 microm in diameter. No re-differentiation into elementary bodies (EBs) was detected. In re-infection experiments, co-infected cells produced fewer EBs than monoinfected cells.
In the present study we confirm that PEDV co-infection alters the developmental cycle of member species of the family Chlamydiaceae, in a similar manner to other well-described persistence induction methods. Interestingly, this effect appears to be partially species-specific as Chlamydia pecorum appears more sensitive to PEDV co-infection than Chlamydia abortus, as evidenced by TEM and IF observations of a homogenous population of aberrant inclusions in PEDV - Chlamydia pecorum co-infections.
衣原体可引起持续性感染,这与人类和动物的多种慢性疾病有关。衣原体与猪流行性腹泻病毒(PEDV)的混合感染可能导致持续性衣原体感染。为了验证这一假说,本研究建立了细胞培养适应的 PEDV 与衣原体流产亚种或衣原体牛生殖道亚种在 Vero 细胞中的体外双重感染模型。
通过免疫荧光(IF)、透射电子显微镜(TEM)和再感染实验对感染培养物进行了研究。通过 IF,感染衣原体的细胞在 39 hpi 后显示出正常包涵体。衣原体流产亚种的双重感染显示出包括小包涵体(由异常体组成)、中包涵体(由异常体和网状体组成)和正常包涵体在内的包涵体类型的混合。只有在衣原体牛生殖道亚种和 PEDV 的双重感染实验中才能观察到异常包涵体。衣原体流产亚种和衣原体牛生殖道亚种混合感染的 TEM 检查显示,包涵体中存在异常的衣原体,包含类似网状的、多形性的异常体,直径可达 2 微米。未检测到再分化为始体(EBs)。在再感染实验中,共感染细胞产生的 EBs 比单感染细胞少。
本研究证实 PEDV 共感染以类似于其他已充分描述的诱导持续性方法的方式改变了衣原体科成员种的发育周期。有趣的是,这种效应似乎部分具有种属特异性,因为与衣原体流产亚种相比,PEDV 共感染对衣原体牛生殖道亚种的影响更为明显,这可以从 TEM 和 IF 观察到 PEDV-衣原体牛生殖道亚种共感染中均一的异常包涵体种群得到证明。