| Project/Area Number |
10671138
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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 |
General surgery
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| Research Institution | Department of Kidney Center(Surgery), Tokyo Women's Medical University |
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Principal Investigator |
HAYASAKA Yuutarou Department of Surgery (Kidney Center), Lecturer, 医学部, 講師 (30120033)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Osame Department of Hematology, Lecturer, 医学部, 講師 (30167712)
FUJITA Shougo Department of Surgery (Kidney Center), Assistant, 医学部, 助手 (58257020)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | anti-αβTCR・autoantibodies / anti-OKT 3・2nd antibodies / FITC-BMA 031 (αβTCR framework) / Auto-MLR / 抗αβTCR・自己抗体 / 自己リンパ球混合反応(Auto-MLR) / OKT 3特異的αβTCR / T細胞クローン(同一αβTCR・T細胞集団) / BMA 031(αβTCR・framework)mAb / T細胞クローン(同-αβTCR・T 細胞集団) / OKT 3投与血清 / Auto-MLR / T細胞クローン / BMA 031・抗体 / OKT 3 / BMA 031(αβTCR・framework) / TCRVβ / 抗ヒトαβTCR・イディオタイプ抗体 / OKT 3処理“Stimulator" / OKT3投与患者血清処理“Responder" |
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| Research Abstract |
[Purpose]We report here a basic study on the detection of autoantibodies to the human T cell antigen receptor (aβTCR) in the sera of patients administered OKT 3. The autoantibodies were studied by Flowcytometry(FACS) using FITC-conjugated anti-human IgG antibodies, BMA 031(αβTCR・framework specific), TCRV β(TCR・variable β-ahain specific) monoclonal antibody. OKT 3 was well known as a monoclonal antibody from mouse IgG_<2a> subtype, and it has a specifcity to the human T cell (CD 3). The OKT 3 also has xenoantigenecities to human T cells. Therefore, the anti-2nd autoantibodies to the OKT 3 were detected in the sera of patients adminstered OKT 3. These anti-2nd antibodies inhibited between OKT 3 and human T cells, and the renegative sera of anti-OKT 3・2nd antibodies were tested continued to detected of autoantibodies to the αβTCR.
Autologous mixed lymphocyte reaction (Auto-MLR) was studid to determine the specificites of the autoantobodies to OKT 3.
[Materials and Methods]In the FACS cell-a
… More
nalysis, the sera of patients administered OKT 3 were incubated with human lymphocytes for 60 mineutes at l0℃, and washed twice with medium RPMI 1640. The samples were stained with FITC-anti human IgG antibodies as positive control, and compared with the same T cells pretreated with BMA 031 to determine the specificities to the αβTCR.The cell analysis by FACS was performed at total cell counts (=Total Events) of 10,000〜30,000 and the percent of inhibition was calculated a follows.Events inhibition (%)=(P.C-E)/P.C x 100. (Events P.C : positive control, E : with patient sera). Auto-MLR was also studied to determine the specificities of the auto-antibodies to the αβTCR using OKT 3-treated Stimulator auto-cells (^<OKT3->Stimulator), and both untreated (positive control) and treated responder autocells with the sera of patients administered OKT 3.Auto-MLR was cultured for 3 days at 37℃ in the 5%-CO_2 incubator. 〔Restults〕Although, the positive reaction of FITC-anti human IgG antibodies to the human lymphocytes appereared when treated with the sera of patients administere OKT 3, this positive reaction was inhibited after the same human lymphocytes were pretreated with the BMA 031 mAb, and the positive reaction of FITC-BMA 031 and TCRVβ mAb. against human lymphocytes also inhibited when the lymphocytes were pretreated with the same sera of patients. The reactivity of auto-MLR was remarkably inhibited also after the treatement of responder auto-cells with the sera of patients administered OKT 3. Less
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