感染HaSNPV棉铃虫幼虫的组织病理和免疫组化研究

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本文报道了棉铃虫单核衣壳核多角体病毒(Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus,HaSNPV)感染棉铃虫幼虫的病理时相及HaSNPV多角体蛋白在幼虫组织中表达的免疫组化研究。以5×10~3PFU的HaSNPV出芽病毒粒子(BV)注射4龄初的棉铃虫幼虫,石蜡切片的H.E染色表明,在感染后24h,病理变化不明显;48h后脂肪体、气管组织的细胞核开始肿大,细胞开始变形;72h后脂肪体、气管、真皮细胞核肿大十分明显,组织结构松散;但中肠和肌肉组织未见明显病变;96h后,脂肪体、气管、真皮组织结构完全被破坏,肌肉组织变疏松。免疫组化结果表明,感染后24h,未能检测到多角体蛋白在幼虫组织中的表达;感染后48h,多角体蛋白可在部分脂肪体、气管组织、血细胞和真皮组织的细胞核中表达;72h后,被感染的脂肪体、气管组织、和真皮组织的细胞数比48h多;96h后,多角体蛋白可以在脂肪体、气管、真皮组织中大量表达,48h到96h间,被感染的血细胞数目基本不变,中肠组织和肌肉都未检测到多角体蛋白的表达;幼虫死亡后,可在中肠上皮细胞的基底膜间隙中检测到多角体蛋白的表达,肌肉组织中未见表达信号,其它组织全部被感染并且组织结构被破坏,H.E的结果与免疫组化的结果基本相符。 In this paper, the pathological phases of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) infected Helicoverpa armigera larvae and the immunohistochemical study of HaSNPV polyhedrin expression in larval tissues were reported. The 4th instar H. armigera larvae were injected with 5 × 10 ~ 3 PFU of HaSNPV budding virus particles (BV). HE staining of paraffin sections showed that the pathological changes were not obvious at 24 h after infection. After 48 h, the nuclei of fat body and trachea began Swollen, the cells began to deform; 72h fat body, trachea, dermis nuclei were obvious swelling, loose tissue structure; but no obvious lesions in the midgut and muscle tissue; 96h, fatty body, trachea, dermal tissue structure completely destroyed Muscle tissue becomes loose. The result of immunohistochemistry showed that the expression of polyhedrin in larval tissues could not be detected 24 h after infection. At 48 h after infection, polyhedrin was expressed in the nuclei of some fat bodies, trachea, blood cells and dermis tissues. After 48h, the number of cells in infected fat body, trachea and dermis was more than that in 48h. After 96h, polyhedrosis protein could be expressed in fat body, trachea and dermis, and the number of infected blood cells between 48h and 96h Polyhedrin expression was not detected in midgut tissue and muscle. After larval death, polyhedrin expression was detected in the basement membrane space of midgut epithelial cells, no expression signal was found in muscle tissue, All other tissues were infected and their structures were destroyed. The results of HE basically matched the results of immunohistochemistry.
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