| Project/Area Number |
09670871
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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 |
Dermatology
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| Research Institution | YAMANASHI MEDICAL UNIVERSITY |
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Principal Investigator |
SHIMADA Shinji Yamanashi Medical University, School of Medicine, Professor., 医学部, 教授 (10114505)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIYAMA Hideaki Yamanashi Medical University, School of Medicine, Assistant Staff., 医学部, 助手 (70301207)
長田 厚 山梨医科大学, 医学部, 助手 (60262656)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | SLAM / co-stimulation / T cell activation |
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| Research Abstract |
SLAM is constitutively expressed on memory T cells, a part of B cells, immature thymocytes, but not on naive T cells. However, SLAM is rapidlyex pressed on naive T cells when activated. It is also known that SLAM on T cells has a co-stimulation activity by crosslinking stimulation with agonistic mAb.
To analyze the biological role of SLAM, especially for mechanism of the input of co-stimulation by SLAM, we tried to identify the ligand of SLAM.For the purpose, we subcloned the cDNA of SLAM by RT-PCR from RNA purified from the PHA-stimulated PBT, and made the new construct (plasmid) of fusion protein composed with extracellular portion of SLAM(lto+717) and the cDNA of Fc portion of human IgG I by inserted into pCDM8. These plasmid (pCDM8-SLAM-Ig) was stably transtected into P3UI cells, and we purified much of SLAM-Ig fusion protein from supematants ofthese cells. We also generated and purified CTLA4-Lg and ICAM- 1-Ig fusion proteinswith similar techniques. We investigated the importance of the interaction between these accessory molecules in T cell activation mediated by antigen presenting cells, using these fusion proteins. Although CTLA4-Ig or ICAM-l-Ig could inhibit the T cell activation by blocking the interaction of CD28/CD86 or ICAM- l/LFA1, SLAM-Ig could not affect on T cell activation in our system. We also tried to stain various cells with SLAM-Ig, none of cells were stained. We speculated the cause of the disfunctionof our SLAM-Ig as making bimer by the disulfide-binding in the portion of hinge. We are now generating a new construct which does not have disulfidate portion in hinge. This might be more useful for our purposes.
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