| Project/Area Number |
07670226
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human pathology
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| Research Institution | AIDS RESEARCH CENTER,NATIONAL INSTITUTE OF HEALTH |
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Principal Investigator |
SATA Tetsutaro NATIONAL INSTITUTE OF HEALTH,AIDS RESEARCH CENTER,HEAD, エイズ研究センター, 室長 (00162397)
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| Co-Investigator(Kenkyū-buntansha) |
KURATA Takeshi NATIONAL INSTITUTE OF HEALTH,DEPARTMENT OF PATHOLOGY,DIRECTOR., 感染病理部, 部長 (50012779)
IWAQSAKI Takuya NATIONAL INSTITUTE OF HEALTH,DEPARTMENT OF PATHOLOGY,HEAD., 感染病理部, 室長 (90146027)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Human cytomegalovirus / IEl protein / anti-lE antibodies / Cytomegalovirus pneumonia / Retinal pigment epithelial cell. / 前初期遺伝子 / 単クローン抗体 |
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| Research Abstract |
The aim of this research project is to develop the methods, or useful antibodies and probes to analyze the localization and the stage of human cytomegalovirus (HCMV) infection in the fixed tissues taken mainly from antopsy cases. The HCMV-infected cells, furthermore, are examined for their characterization. Taken together, the pathogenesis of the lesion infected with HCMV in organs is examined.
Since several immediate-early proteins of HCMV have been identified so far, we cloned exon 4 and 5 of immediate-early gene using PCR method in order to differentiate the products of IE1 and IE2 genes, and expressed them in the form of the Glutathion S transferase fusion protein in E.coli system. Western blot analysis using the fusion proteins revealed that anti-IE2 monoclonal antibody (E5) idenified C-terminal peptide encoded by exon 5. Anti-IE1 rabbit antibody was also established, and cold detect sensitively the early phase of the infected cells which did not show a characteristic sytomegalic cells in the tisses. E5 was also available on the sections treated with microwave beforehand. These antibodies could demonstrate many HCMV infected cells without characteristic features in the interstitial area of the lung. Using lung tisses taken from AIDS autopsy cases we found the true HCMV infection necessary for the treatment if the infected cells were located in the interstitium of the alveolum in the lung. We also analyzed the status of the viral proliferation in the retinal pigmented epithilial cell line, K-1034, using antibodies and phobes described, and found the differences of virus proliferations in RPE from those in human embryonic lung fibroblasts and the serum factors were considered to influence the HCMV proliferation especially in the epithelial cells. We continue these studies using RNA in situ hybridization methods with IE genes and late genes of HCMV.
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