| Project/Area Number |
01480255
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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 |
Pediatrics
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| Research Institution | Tohoku University |
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Principal Investigator |
TSUCHIYA shigeru Research Institute for Tuberculosis and Cancer・ Department of Pediatrics・ Associate Professor, 抗酸菌病研究所, 助教授 (30124605)
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| Co-Investigator(Kenkyū-buntansha) |
MASAYOSHI Minegishi Research Institute for Tuberculosis and Cancer. Department of Pediatrics・Joshu, 抗酸菌病研究所, 助手 (20211592)
SATO Tetsuo Research Institute for Tuberculosis and Cancer・ Department of pediatrics・ Joshu, 抗酸菌病研究所, 助手 (90170761)
KONNO Tasuke Research Institute for Tuberculosis and Cancer・ Department of Pediatrics・ Profes, 抗酸菌病研究所, 教授 (00004846)
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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 |
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥4,200,000 (Direct Cost: ¥4,200,000)
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| Keywords | X linked agammaglobulinemia / Clonal transformation / Epstein-Barr virus / IgD producer / kappa-lambda double producer / B cell growth factor / エプスタイン・バ-ウィルス / 無ガンマグロブリン血症 / エプスタイン-・バ-ウィルス / 未熟B細胞株 / K-λダブルプロデュ-サ- |
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| Research Abstract |
Immature B lymphoblastoid cell lines (B-LCL) were established from bone marrow blood of a patient with X-linked agammaglobulinemia (X-LA). One of the characters of those LCLs is poor growth capability in in vitro culture. However, we established K4-6-22 cells from K4 parent cells with vigorous in vitro growth capacity. The cells can make colonies with 1.9% of plating efficiency and adapt serum free medium very easily. Demonstration of B-cell stimulatory activity in the culture supernatant of K4-6-22 suggested the autocrine mechanism of the vigorius growth of the cells. Both K4 and K4-6-22 cells express double light chains, that is kappa and lambda, with single mu heavy chain. Repeated cloning of K4-6-22 in semisolid agarose always show both kappa and lambda light chains in each clone. These data indicates the simultaneous expression of kappa and lambda light chains in a single clone. However, we were not able to establish cloned cells which continuously express both ligh chains as well as mu heavy chain. Another interested cell line is a K5 which produce delta heavy and lambda light chain. Cell sorting using flow cytometer and successive cloning in semisolid agarose, we established the cloned cell line, K5S. But we did not find any IgD production in the cell line. After thawing, immunoprecitation experiments of original K5 cells using monoclonal anti-delta and lambda antibodies were carried ort and real 62 KDa delta and 32 KDa lambda proteins were demonstrated. Using immunoglobulin J_H probe we performed southern blot analysis of DNA of B-LCL from a X-LA patient an demonstrated clear rearranged bands in all cells including K5, K5S, K4 and K4-6-22.
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