Garrido-Fariña G I, Cornejo-Cortés M A, Martínez-Rodríguez A, Reyes-Esparza J, Alba-Hurtado F, Tórtora-Pérez J
Department of Biological Sciences, Morphological Sciences Section, Support Laboratory for Histology and Biology, FES-Cuautitlán UNAM, Mexico.
Vet Microbiol. 2008 May 25;129(1-2):28-39. doi: 10.1016/j.vetmic.2007.11.009. Epub 2007 Nov 19.
Contagious ecthyma virus (CEV) is a disease caused by a parapoxvirus, also is a potent genetic carrier with the capacity for regulating apoptosis in the cells of infected skin, a mechanism that serves for evading the immune response of the host. It has been suggested that the virus may remain in the skin and be able to cause repeated infections in the same flock. The effect of infection as well as the presence of contagious ecthyma virus was evaluated in terms of lesions and apoptosis in the skin of animals, infected both naturally and experimentally. Samples used were obtained from a naturally infected sheep, 5 goats inoculated with CEV and a negative control. Samples obtained were longitudinally sectioned and processed using photon and electron microscopy, and embedded in paraffin and araldite. Samples embedded in paraffin were sectioned in 5 microm of thickness and dyed with orange eosin-hematoxilin G and Gomori's trichrom stain, apoptosis was demonstrated by the TUNEL assay, the viral antigen was revealed using polyclonal antibodies, and the presence of lymphocytes CD4+ and CD8+, with monoclonal antibodies. The samples processed in resin were cut to obtain semi-fine sections and dyed with toluidine blue-borax, and the ultra-fine sections were impregnated with lead citrate and uranyl acetate. Observations were similar in both, the natural infected animal and the experimental group. Infiltration was observed as well as images suggestive of a process of apoptosis. The TUNEL assay demonstrated that the number of epithelial cells undergoing apoptosis diminished during the process and increased among defense cells, until they almost disappeared at the beginning of healing. Cells undergoing apoptosis were located near the sebaceous glands and pilose follicles. The infiltrated lymphocytes gradually diminished. The viral antigen was observed in cells with morphology suggestive of apoptosis, located in sebaceous glands and pilose follicles. Using electron microscopy, cells with morphology compatible with that of lymphocytes were observed to be undergoing apoptosis, but there was little evidence of viral particles.
传染性脓疱病毒(CEV)是由副痘病毒引起的一种疾病,也是一种强大的基因载体,具有调节受感染皮肤细胞凋亡的能力,这种机制有助于逃避宿主的免疫反应。有人认为该病毒可能残留在皮肤中,并能够在同一羊群中引起反复感染。通过对自然感染和实验感染动物皮肤中的病变和凋亡情况进行评估,来研究感染的影响以及传染性脓疱病毒的存在情况。所使用的样本取自一只自然感染的绵羊、5只接种了CEV的山羊以及一个阴性对照。获取的样本进行纵向切片,并使用光学显微镜和电子显微镜进行处理,然后分别包埋在石蜡和环氧树脂中。包埋在石蜡中的样本切成5微米厚的切片,用橙色伊红 - 苏木精G和Gomori三色染色法染色,通过TUNEL检测法显示凋亡情况,使用多克隆抗体揭示病毒抗原,并用单克隆抗体检测CD4 +和CD8 +淋巴细胞的存在情况。用树脂处理的样本切成半薄切片,用甲苯胺蓝 - 硼砂染色,超薄切片用柠檬酸铅和醋酸铀进行浸渍。在自然感染动物和实验组中观察到的结果相似。观察到有浸润现象以及提示凋亡过程的图像。TUNEL检测表明,在这个过程中发生凋亡的上皮细胞数量减少,而在防御细胞中增加,直到在愈合开始时几乎消失。发生凋亡的细胞位于皮脂腺和毛囊附近。浸润的淋巴细胞逐渐减少。在皮脂腺和毛囊中具有提示凋亡形态的细胞中观察到病毒抗原。使用电子显微镜观察到具有与淋巴细胞形态相符的细胞正在发生凋亡,但几乎没有病毒颗粒的证据。