Detection of autoantibodies to the T cell antigen receptor (αβTCR) in the sera of patients administered OKT 3.

检测接受 OKT 3 的患者血清中 T 细胞抗原受体 (αβTCR) 的自身抗体。

• 批准号: 10671138
• 负责人: HAYASAKA Yuutarou
• 金额: $ 2.05万
• 依托单位: Department of Kidney Center(Surgery), Tokyo Women's Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671138/
• 关键词: 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

[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

[目的]我们在这里报告了一项基础研究，该研究在使用OKT 3的患者血清中对人T细胞抗原受体（AβTCR）的检测进行检测。使用FITC-CONCONCONCONTRY（FACS）使用FITC-偶联的抗抗人类IgG抗体，BMA 031（a）（FACS）研究自身抗体。 β（TCR ・变量β-ahin特异性）单克隆抗体。 OKT 3被称为小鼠IgG_ <2A>亚型的单克隆抗体，并且具有对人T细胞的特异性（CD 3）。 OKT 3还具有对人T细胞的异种抗固醇。 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 studiod to determine the specifics of the autoantobodies to OKT 3.[Materials and Methods]In the FACS cell-a … More analysis, 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,并与用BMA 031预处理的相同T细胞相比，以确定αβTCR的规格。FACS通过总细胞计数（=总事件）进行10,000-30,000的细胞分析，并计算了抑制百分比。events抑制（％）=（p.c-e）=（P.C-E）/p.c.c x 100。还研究了自动-MLR，以确定使用OKT 3处理的刺激器自动细胞（^<okt3->刺激器）确定自动抗体对αβTCR的规格，并且未经治疗（阳性对照）和未经治疗的呼应和对呼应的患者均可使用3.auto-ml.t Atty-ianto-mlr-mlr-mlr-mlr进行了治疗。孵化器。 [恢复]尽管FITC-anti人IgG抗体对人淋巴细胞的阳性反应在用患者的血清治疗3时出现，但在用BMA 031 MAB和FITC-BMA 0331和TCRV的阳性反应中，将同样的人淋巴细胞抑制了这种阳性反应。当淋巴细胞用相同的患者血清预处理时，针对人淋巴细胞也会抑制。在用OKT 3的患者的血清治疗响应者自动细胞后，自动MLR的反应性也受到明显抑制。

项目成果

早坂勇太郎 他: "ヒト血清中の抗ブタ赤血球凝集・阻害抗体と抗TCR・自己抗体の検討"第2回日本異種移植研究会抄録集,. 2. 17 (1999)

Yutaro Hayasaka 等：“人血清中的抗猪血凝/抑制抗体和抗 TCR/自身抗体的研究”，第二届日本异种移植研究会摘要，2. 17 (1999)

早坂勇太郎,他: "ヒト血清中の抗ブタ赤血球凝集・阻害抗体と抗TCR・自己抗体の検討"第2回日本異種移植研究会抄録集,. 2. 17 (1999)

Yutaro Hayasaka 等：“人血清中抗猪血凝/抑制抗体和抗 TCR/自身抗体的研究”，第二届日本异种移植研究组摘要，2. 17 (1999)

早坂勇太郎 他: "ヒト血清中の抗ブタ赤血球凝集・阻害抗体と抗TCR・自己抗体の検討."第2回日本異種移植研究会(同抄録集). 2. 17 (1999)

Yutaro Hayasaka 等：“人血清中的抗猪血凝/抑制性抗体和抗 TCR/自身抗体的研究”，第 2 届日本异种移植研究会（摘要集）。

早坂勇太郎 他: "BMA 031投与後の患者血清中の抗-3次抗体とその抗体特異性の検討,"移植. (掲載予定). (2001)

Yutaro Hayasaka 等人：“BMA 031 给药后患者血清中抗三级抗体的检查及其抗体特异性”，《移植》（待出版）。

Yuutarou Hayasaka, et al: "Basic study on the specificities of anti-3rd auto -antibodies to the BMA 031 in the sera of patients administered BMA 031 monoclonal antibody. The 36th Annual Meeting of the Transplantation society of Japan (2000)"Japanese Journ

Yuutarou Hayasaka 等人：“在给予 BMA 031 单克隆抗体的患者血清中抗第 3 代自身抗体对 BMA 031 的特异性的基础研究。第 36 届日本移植学会年会（2000 年）”日本期刊