| Project/Area Number |
01570271
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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 |
Immunology
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| Research Institution | National Institute of Neuroscience, National Center of Neurology and Psychiatry (1990)
Kochi Medical School (1989) |
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Principal Investigator |
YAMAMOTO Hiroshi Nat'l. Inst. Neurosci., Nat'l. Ctr. Neurol. Psych., Div. Neuroimmunol., Chairman, 神経研究所, 部長 (50127312)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIMOTO Shigeyoshi Kochi Med. Sch., Dept. Immunol., Professor, 医学部, 教授 (00009151)
谷口 武利 高知医科大学, 医学部, 助教授 (90127944)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Idiotype / Idiotype-network / B lymphocytes / Long-term culture / Genetic restriction / Immunoglobulin gene / 遺伝子拘束性 / マウスB細胞 / B細胞クロン / B-B細胞間相互作用 |
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| Research Abstract |
Immunization of BALB/c mice with MOPC104E myeloma protein induces antiidiotypic B lymphocytes that have idiotype-specific enhancing activity on antibody production. The B-B cell interaction was restricted to both Igh and class II MHC. However, anti-Thy-1 and complement treated splenic B cells were maintained for more than 1 year in a mixture of concanavalin-A-stimulated splenocyte culture supernatand and synthetic medium. In applying the long term culture method, we have established a cloned B cell line named B19-1^d. B19-1^d cell are specific to MOPC104E or J558 cross-reactive idiotype and they express surface mu, lambda but no Ly-1. B19-1^d do not spontaneously secrete immunoglobulin but produce them upon stimulation with bacterial lipopolysaccharides. The effect of B19-1^d cell line on idiotypic antibody production was tested. Addition of only 10 to 100 B19ー1^d cells into dextran immune B cell culture greatly enhanced the idiotype-positive antidextran antibody responses. On the contrary, the antidextran antibody production was suppressed by the higher doses of B19-1^d cells. The effective cooperation between dextran-immune B cells and B19-1^d cloned B cells was restricted to class II MHC. By fusing lipopolysaccharide-stimulated B19-1^d clone with P3U1 myeloma, antiidiotypic antibody producing hybridoma was established.
Each hybridoma cDNA and MOPC104E genomic DNA was cloned and their nucleic acid sequences were determined. The sequences were highly similar each other (91% similarity) and it was suggested that the both genes were derived from closely linked germ line genes.
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