| Project/Area Number |
02454248
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | The Kitasato Institute |
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Principal Investigator |
ONO Akira The Kitasato Institute Department of Virology, Associate Director, ウイルス部, 室長 (80160901)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Tetsu Tokyo Metropolitan Ohtsuka Hospital Department of Internal Medicine, 内科, 医長
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | myasthenia gravis / thymus / thymoma / retrovirus / 遺伝子クロ-ニング / レトロウィルス |
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| Research Abstract |
By electron microscopy, we have demonstrated in retrovirus particles in cultured thymus cells (thymomas and thymus hyperplasias) of patients with myasthenia gravis. In most cases, these particles were detected in thymus epithelial cells when cultured with mitomycin C-treated human B-cells. For further characterization of these particles, virus-producer cell lines were established by co-culturing human T-lymphoblastoid cells with B-cell-stimulated human thymus cells. Mn^<++>-dependent RNA- directed DNA polymerase activity was detected at a density of 1.15-1.17 g/cm^3 on sucrose density gradients in the culture fluid of these co-cultured T-cells. We molecularly cloned the cDNA prepared by the detergent-activated endogenous polymerase reaction in the culture fluid of KK-ll cells, one of the virus-producer T-cell lines. The ^<32>p-labelled cloned cDNA hybridized to DNAs from freshly prepared hyman thymoma cells, in addition to KK-ll cell DNA. We sequenced about 400-1200 bp of seven cDNA clones, and the sequence of each cDNA was compared with that of all genes with known sequences present in EMBL data base. No striking similarity was observed when compared to other sequences. The nucleotide sequence of seven cloned cDNAs constantly revealed high homology (50-70%) with that of retroviruses such as HIV-l and equine infectious anemia virus (EIAV). These results indicate that cDNAs obtained here are new viral genes, possibly retroviral genes, derived from human thymus cells.
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