IMMUNOLOGICAL AND IMMUNOCHEMICAL ANALYSIS OF HUMAN INTERLEUKIN 1 AND ITS RECEPTOR
| Project/Area Number |
60570225
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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 | JICHI MEDICAL SCHOOL |
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Principal Investigator |
KASAHARA TADASHI JICHI MEDICAL SCHOOL, ASSOCIATE PROFESSOR, 医学部, 講師 (60049096)
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| Co-Investigator(Kenkyū-buntansha) |
MUKAIDA NAOFUMI JICHI MEDICAL SCHOOL, ASSISTANT, 医学部, 助手 (30182067)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1986: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1985: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Interleukin 1 (IL1) / Monoclonal antibody / Western blotting / IL1 receptor / シグナル伝達機構 |
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| Research Abstract |
In order to promote the study on the immunological function and immunochemical prop erties of interleukin 1 (IL1) and its receptor, it is prerequisite to obtain purified materials and highly specific antibodies (Ab) against those materials. Fortunately in Japan, two industories, Dainippon and Ohtuka Pharmaceutical Companies have recently been successful in the gene cloning of human IL1 <alpha> and IL1 <beta> , respectively, so that recombinant IL1 products are available to us. Therefore, we attempted to raise monoclonal and polyclonal Ab against human IL1 <alpha> , and we could get anti-IL1 <alpha> Ab which had strong neutralizing activity against human IL1 <alpha> species with high specificity. Those data have been substantiated by the neutralization of IL1 from various sources, and western blot analysis. In addition, those Ab have been utilized for 1) the determination of IL1 from various cell sources; 2) separation of IL1 subtype utilizing as Ab conjugated columns; 3) establishement of ELISA for the quantitative determi tion of IL1 subtype in various body fluids.
Subsequently, we exploired the IL1 <alpha> (or <beta> ) receptors on various cells using <^(125)I> -labeled IL1. We found high affinity IL1 <alpha> (or <beta> ) receptors on NK-like YT cells with -4x <10^3> receptors/cell. However, we detected only low levels of IL1 receptor on human leukemic HSB.2 cell clones with less than <10^2> /cell, which produce high levels of IL2 in an IL1-dependent manner. Such IL1-dependent IL2 production phenomena are now being actively investigated in the context of signal transduction transmitted by IL1 as an important conseqent event occurring after IL1 binding to the cell receptor.
A part of our results have been accepted by the Journal of Immunology (in press, 1987) and other part is being submitted.
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Report
(1 results)
Research Products
(14 results)
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[Publications] Ohike,Y., Imai,M., Tanaka,E., Mukaida,N., Kasahara,T.,et al.: "A RADIOIMMUNOASSAY THAT SANDWICHES HUMAN INTERLEUKIN 2 BETWEEN RADIOLABELED MONOCLONAL ANTIBODY AND THE RECEPTOR ON A HEMATO POIETIC CELL LINE" JOURNAL OF IMMUNOLOGICAL METHODS. 87. 245-249 (1986)
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