| Project/Area Number |
63440018
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Osaka City University |
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Principal Investigator |
KINOSHITA Yoshihiro Osaka City University, Medical School, Professor, 医学部, 教授 (80046896)
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| Co-Investigator(Kenkyū-buntansha) |
KITAMURA Kenji Osaka City University, Medical School, Research Associate, 医学部, 助手 (40214811)
TOMINAGA Keiko Osaka City University, Medical School, Research Associate, 医学部, 助手 (70217545)
TERANO Yoshitake Osaka City University, Medical School, A-Part-Time Lecturer, 医学部, 非常勤講師
HATO Fumihiko Osaka City University, Medical School, Lecturer, 医学部, 講師 (00198772)
HUNAE Yoshihiko Osaka City University, Medical School, Professor, 医学部, 教授 (00047268)
豊川 奉 大阪市立大学, 医学部, 助手 (90172741)
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| Project Period (FY) |
1988 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥32,500,000 (Direct Cost: ¥32,500,000)
Fiscal Year 1991: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1989: ¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1988: ¥23,000,000 (Direct Cost: ¥23,000,000)
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| Keywords | Thymus epithelial cell / Biologically active factor / Synthesis and secretion / Neurotransmitter / Endcrine hormones / Thymic lymphocytes / Differentiation and maturation / Monoclonal antibody / ラット胸腺上皮性細胞 / 培養上清 / 胸腺ホルモン / 高速液体クロマトグラフィ- / 分離・精製 / 神経刺激伝達物質 / 神経・内分泌系 / 分泌調節 / ポリペプチドの分離・精製 / T細胞の分化・成熟 / 生体防衛機構の個体発生と老化 / ポリベブチドの分離・精製 |
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| Research Abstract |
(1) Separation of biologically active polypeptides (BAPs) from the culture supernatant of thymus epithelial cells (TEC).
We tried to separate BAPs from the culture supernatant of TEC line, IT-45R1. The polypeptide fraction with 2.5kD in molecular weisht possessed the capacity to induce the responsiveness of rat thymic lymphocytes (TLs) against concanavalin A and phytohemagglutinin. By further separation using reversed-phase column, the former was found in the hydrophobic fraction and its target cells were enriched in lighter TLs, while the latter was detected in the hydrophilic fraction and its target was heavier TLs. About 80% of tat TLs bind to guinea pig erythrocytes and from rosettes (RFC), whereas other TLs (non-RFC) do not. The 3kD fraction possessed the capacity to endow non-RFC with resette forming activity. Furthermore, rosette formation-inducing activity was found in the two different fractions separated by reversed-phase column method. By the same methods described above, the
… More
activity to induce proliferation of TLs was recognized in 10kD fraction from the culture supernatant of TAD3, derived from lymphocyte dominant thymoma. It is assumed that the target cells for 10kD fraction bear CD4-8- phenotype, observed in the most immature TLs and that the adherent cells in thymus take, also, an role in the proliferation system.
(2) Effects of neurotransmitter and hormone on the function of thymus epithelial cells.
The effect of cholinergic agonists on TEC was examined by using TAD3. It is revealed that cholinergic agonists stimulate TEC to proliferate at preconfluent state. On the other hand, at confluent state, the agonists stimulate TEC to synthesize proteins. These activities were possibly induced by the signal through nicotinic acetylcholine receptor on TEC surface. Furthermore, the effect of growth hormone on TEC was examined. It is suggested that the hormone enhances the mitotic index of the TAD3 cells at preconfluent state and enhances the protein synthesis of IT-45R1 cells at confluent. Furthermore, growth hormone is detected to increase thymosin alpha 1 aynthesis of IT-45R1 cells at confluent state by using anti-synthetic thymosin alpha 1 antibody, prepared by us. Less
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