权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

RESEARCH ON MECHANISNS OF HSV-1 LATENT INFECTION BY IN SITU HYBRIDIZATION

原位杂交研究HSV-1潜伏感染机制

基本信息

批准号：
62570356
负责人：
NAKAMURA Shozo
金额：
$ 1.41万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570356/
关键词：
Herpes simplex encephalitis antigens nucleic acid sequences 核酸免疫組織化学法 in situ hybridization法 in situ ハイブリダイゼーション

项目摘要

In order to elucidate the correlation between herpes simplex virus(HSV-1) and the central nervous system tissue, we performed the detection of viral antigens and RNA in the brain tissue sections from a patient with herpes simplex virus encephalitis using immunocytochemistry and in situ hybridization.In the present study the hybridization protocol reported by Haase AT et al. were applied for the detection of viral RNA and antigens with a few modifications.The sections were first immunocytocehmically stained to detect HSV-1 antigens by ABC method, and then hybridized with ^3H-labelled HSV-1 cDNA probe for the detection of RNA after the acetylation of sildes for the prevention of nonspecific bibdings of isotope to slides.HSV-1 infacted Vero cells as positive control demonstrated both antigens and RNA. However, no hybridization signals and color deposits were observed in uninfacted Vero cells.In frozen brain tissue sections of herpes simplex encephalitis the viral antigen(VA) positive cell … More s, containg brown-colored deposits located primarily in the cytoplasm, were widely distributed throughout the lesions of a part of gray matter. Such VA positive cells were morphologically identified to be swollen neurons, glial cells and endothelial cells. In addition, some of VA positive cells have also hybridization signals over the nuclei, namely VA and RNA positive cells which were diffusely located colse to VA positive cells. Moreover, only RNA postive cells with numerous silver grains over the nuclei were predominantly distributed at the neighborhood of VA positive cells and at the white matter.In quantitative analysis of HSV-1 infecgted cells of a part of gray matter, cells positive with viral antigen were counted 23.2% of total cells.Both VA and RNA cells and only RNA cells were detected 13.4% and 18.3% respectively. In addition, in a part of white matter VA and RNA cells and only RNA cells were found 9.3% and 41.7% although there were was not detected VA positive cells.We concluded that the herpes simplex virus (HSV) infects the virious kind of cells composed of brian tissues and shows the virious stages of infection, namely; VA positive cells, VA and Rna positive cells and only RNA positive cells. Moreover, it may be considered that these various stages of infacted cells are developed according to many factors such as different kind of cells, infactious routes and host immune responses. Less

为了阐明单纯疱疹病毒(HSV-1)与中枢神经系统组织之间的相关性，我们采用免疫细胞化学和原位杂交技术对一名单纯疱疹病毒脑炎患者脑组织切片中的病毒抗原和RNA进行了检测。本研究采用Haase AT等人报道的杂交方案。经少量修改后用于病毒RNA和抗原的检测。切片首先进行免疫细胞化学染色，采用ABC法检测HSV-1抗原，然后与^3H标记的HSV-1 cDNA探针杂交，在乙酰化后检测RNA，以防止同位素的非特异性结合幻灯片。HSV-1 感染的 Vero 细胞作为阳性对照，显示出抗原和 RNA。然而，在未感染的Vero细胞中未观察到杂交信号和颜色沉积。在单纯疱疹脑炎的冰冻脑组织切片中，病毒抗原（VA）阳性细胞主要位于细胞质中，含有棕色沉积物，广泛分布于部分灰质的病灶中。此类VA阳性细胞经形态学鉴定为肿胀的神经元、神经胶质细胞和内皮细胞。此外，一些VA阳性细胞在细胞核上也有杂交信号，即VA和RNA阳性细胞，它们广泛位于VA阳性细胞附近。此外，只有核上有大量银粒的RNA阳性细胞主要分布在VA阳性细胞附近和白质处。在对部分灰质的HSV-1感染细胞进行定量分析时，病毒抗原阳性细胞占总细胞的23.2%。VA和RNA细胞均检出，仅RNA细胞检出分别为13.4%和18.3%。此外，在部分白质VA和RNA细胞中，虽然没有检测到VA阳性细胞，但仅发现了RNA细胞，分别占9.3%和41.7%。 VA 阳性细胞、VA 和 Rna 阳性细胞以及仅 RNA 阳性细胞。此外，可以认为，感染细胞的这些不同阶段是根据不同类型的细胞、感染途径和宿主免疫反应等多种因素而发展的。较少的