Senderskiĭ I V, Pavlova O A, Timofeev S A, Dolgikh V V
Parazitologiia. 2014 Jan-Feb;48(1):63-70.
Immunolabeling method of microsporidium Paranosema locustae proteins on cryosections of locust infected fat body was proposed. In contrast to single parasite cells and artificially infected host cell cultures, this method allows to study molecular mechanisms of host-parasite relationships and in particular the secretory microsporidial proteins either at cellular or tissue level. Immunolocalization of the EPR-specific and cytoplasmic forms of Hsp70 family of molecular chaperones on cryosections showed accumulation of these proteins in the respective compartments of intracellular developmental stages of P. locustae and their absence in host structures. This allows to use them in diagnostics of microsporidiosis lesions in infected tissues as well as in colocalization analysis with P. lociustatre secretory proteins as a marker of parasite. The cytoplasmic chaperone stains cytoplasmic compartment homogeneously, but in the infected host cell during sporogony it disappears partially from the intracellular stages of development which damaged by maturing spores. Thereby study of molecular mechanisms of host-parasite relationships is to be carried out at the earlier stages of infection before active sporogony.
提出了在感染蝗虫的脂肪体冷冻切片上对蝗虫微孢子虫蛋白质进行免疫标记的方法。与单个寄生虫细胞和人工感染的宿主细胞培养物不同，该方法能够在细胞或组织水平上研究宿主 - 寄生虫关系的分子机制，特别是微孢子虫分泌蛋白。分子伴侣Hsp70家族的EPR特异性和细胞质形式在冷冻切片上的免疫定位显示，这些蛋白质在蝗虫微孢子虫细胞内发育阶段的相应区室中积累，而在宿主结构中不存在。这使得它们可用于诊断感染组织中的微孢子虫病病变，以及与蝗虫微孢子虫分泌蛋白进行共定位分析，作为寄生虫的标志物。细胞质伴侣均匀地染色细胞质区室，但在孢子形成过程中感染的宿主细胞中，它在被成熟孢子破坏的细胞内发育阶段会部分消失。因此，宿主 - 寄生虫关系的分子机制研究应在活跃孢子形成之前的感染早期阶段进行。
