1994 Fiscal Year Final Research Report Summary
Epstein-Barr virus mutagenesis by homologous recombination
| Project/Area Number |
05670286
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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 |
Virology
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| Research Institution | Nihon University |
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Principal Investigator |
FUJIWARA Shigeyoshi Nihon University, School of Medicine, Lecturer, 医学部, 講師 (30173488)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Yasushi Nihon University, School of Medicine, Professor, 医学部, 教授 (30004675)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Epstein-Barr Virus / Homologous Recombination / Recombinant Virus / Hygromycin / MT-2 / BZLF1 / Persistent Infection |
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| Research Abstract |
The primary goal of this project is to generate Epstein-Barr virus (EBV) recombinants lacking the function of the EBNA3C gene and to characterize their biological activities. A targeting vector pTE3C-Hyg was constructed to insert a hygromycin resistance gene into the EBNA3C gene. This vector was transfected to the EBV-producer line B95-8 and the recombinant viruses released from the transfected B95-8 cells were inoculated to either a human EBV-negative B-cell line BJAB or cord blood mononuclear cells. Recombinant viruses were recovered in either BJAB subclones or lymphoblastoid cell lines that grew in the presence of hygromycin. The viruses recovered in these BJAB clones were shown to be the products of homologous recombination in the EBNA3C gene, whereas those recovered in lymphoblastoid lines were shown to be random recombinants. We are currently characterizing the biological activities of these recombinant viruses.
We used EBV recombinants also as a tool to generate a new experimental system of persistent EBV infection. Infection of the human T-cell line MT-2 with these EBV recombinants and subsequent selection with hygromycin permitted isolation and long-term maintenance of EBV-infected MT-2 clones. The latent EBV gene expression in these EBV-infected MT-2 clones was shown to be similar to that observed in peripheral human T-cell lymphomas. Thus these clones could be useful as a model of EBV gene regulation in T-cell lymphomas. Besides the latent genes, EBV-infected MT-2 clones constitutively express the BZLF1 gene, an immediate-early gene that triggers the productive viral cycle in B lymphocytes. Since EBV production was not detected in these MT-2 clones, we speculate that EBV gene regulation may be different between B- and T-lymphocytes.
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[Publications] Nagatsuka, Y., Watarai, S., Yasuda T., Higashi, H., Yamagata, T., and Ono, Y.: "Production of human monoclonal antibodies to the i blood group antigen by EBV-induced transformation : possible presence of a new glycolipid in cord red cell membrane and human hematopoietic cell lines" Immunology Letters. 46. 93-100 (1995)
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[Publications] Nagatsuka, Y., Hanazawa, S., Kuroiwa, Y., Fukuda, T., Suganuma, T., and Ono, Y.: "Anti-bacterial antibodies in Epstein-Barr virus (EBV) - transformed oligoclonal B-cell lines established from normal persons and autoimmune disease patients." Lett.Appl.Microbiol.19. 206-209 (1994)
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「研究成果報告書概要(欧文)」より
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