| Project/Area Number |
62570356
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
NAKAMURA Shozo Department of Neurology, Institute of Brain Diseases, Staff, 医学部, 助手 (80108498)
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| Co-Investigator(Kenkyū-buntansha) |
TUKAMOTO Teturo Department of Neurology, Institute of Brain Diseases, Staff, 医学部附属病院, 助手 (20171978)
TAKASE Sadao Department of Neurology, Institute of Brain Diseases, Subprof., 医学部, 助教授 (60004983)
SEKIZAWA Tuyoshi Department of Neurology, Institute of Brain Diseases, Staff, 医学部附属病院, 助手 (50150264)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | 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 |
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| Research Abstract |
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
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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
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