| Project/Area Number |
10670312
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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 |
Immunology
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| Research Institution | Science University of Tokyo,Research institute for Biological Sciences |
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Principal Investigator |
KOZONO Yuko Science University of Tokyo, Research Institute for Biological Sciences, Research Associate, 生命科学研究所, 助手 (30197107)
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| Co-Investigator(Kenkyū-buntansha) |
AZUMA Takachika Science University of Tokyo, Research Institute for Biological Sciences, Professor, 生命科学研究所, 教授 (00028234)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | B cell / Somatic mutation / CD19 / Antigen specific / in vitro |
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| Research Abstract |
In order to study the mechanism of which immuno-globlin gene obtains somatic mutation, we tried to set up in vitro immunization system. First of all, we re-analyze in vivo immunization by our hand. We made sensitive reagents which detect Ag receptor of low affinity (<10ィイD1-6ィエD1) on B cells using avidity. Using these reagents, we could surprisingly detect 2-30% Ag binding B cells of spleen at 7-8 days after immunization. Also, we found CD19 altered upon immunization. We used this conditions that CD19 alters for in vitro immunization system. Next, we cultured treated resting B cells by Ag or a/CD19, allo-reactive T cell line which made during this study, and IL-4 with or without Ag or a/CD19 for 7-10 days. We analyzed frequency of somatic mutation in VH186.2 mRNA by sequencing and found some somatic mutations in especially via CD19 signal upon culture. We could see Ag specific B cells by FACS or ELIspot. Although it is very difficult to distinguish from naturally occurring somatic mutation, since different conditions show different frequency of somatic mutation, it might be said that we show in vitro somatic mutation. Therefore, we can continually analyze immunization system in vitro in detail using this system.
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