Analysis of the T-cell clonality in renal allografts using biopsy specimens.

使用活检标本分析肾同种异体移植物中的 T 细胞克隆性。

• 批准号: 09671649
• 负责人: OBATA Fumiya
• 金额: $ 1.79万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671649/
• 关键词: renal allografts; rejection; biopsy; T-cell receptor; clonality; interferon-gamma; inter leukin-4; Th1 and Th2; 浸潤T細胞; T-cell receptor (TCR); Th1; Th2; TCRαβ; CD4; CD8; TH1; TH2; IL4; PCR; 浸潤細胞; クローナリティー解析; V遺伝子使用頻度; 遺伝子増幅法

In order to characterize the T-cell population participating in renal allograft rejection, I performed several analyses using biopsy specimens. The following results were obtained :1) For the accurate analysis of the T-cell receptor (TCR) diversity using biopsy specimens, I developed a non-radioisotopic micromethod, in which TCR cDNAs amplified by an anchored polymerase chain reaction were hybridized with an array of TCR variable gene DNA fixed on a nylon membrane.2) TCR analysis allowed me to assess the clonality of T-cells participating in renal allograft rejection. The method was particularly useful to monitor the change in clonality after immunosuppressive treatment for the rejection.3) The degree of clonality in renal infiltrating T-cell was not significantly different between acute rejection (AR) ard chronic rejection (CR). However, difference in clonality between infiltrating cells ard that in peripheral blood was larger in AR than CR, indicating that particular T-cel 1 clones accumulated in the allografts receiving AR.4) Virally infected allograft often showed T-cell clonality similar to that of rejected allograft and it was difficult to discriminate them only by clonality analysis.5) The degree of infiltration of alpha-beta T cells, CD4-positive T cells, and CD8-positive T cells in CR allografts were not different from that in AR allografts, indicating that CR, as AR, was mediated by these immune cells.6) The frequency of detection of interferon-gammas in allografts was not significantly different between CR and AR, indicating that Th1 cells participated in both CR and AR.7) Interleukin-4 was detected only in one-third of CR allografts and not in AR allografts, suggesting that some CR were mediated by Th2 cells.

为了表征参与肾脏同种异体移植排斥的T细胞种群，我使用活检标本进行了几次分析。获得了以下结果：1）对于使用活检标本对T细胞受体（TCR）多样性进行准确分析，我开发了一种非放射性病的微方法，其中通过固定的基因DNA固定在nylonmmmbrane上的TCR可变基因dNA杂交的TCR cDNA被锚定聚合酶链反应扩增的TCR cDNA。参加肾脏同种异体移植排斥。该方法对于监测免疫抑制治疗后克隆性的变化特别有用。3）急性排斥（AR）ARD慢性排斥（CR）之间肾脏浸润T细胞的克隆性程度没有显着差异。然而，浸润细胞之间的克隆性差异在AR中的血液比Cr大于Cr，表明在接受AR的同种异体移植物中积累的特定T-CEL 1克隆。 CR移植物中的细胞和CD8阳性T细胞与AR同种异体移植物中的细胞和CD8阳性T细胞没有差异，表明Cr是由这些免疫细胞介导的。6）6）同种异体移植中CR和AR中的干扰素 - Gammas检测频率在Cr和AR之间并没有显着差异，表明所有TH1细胞都在CR.7中均参与了cr.7 cr.7 cr.7 cr.7 innor.7 innots inn.7 innor.7 innor.7 innots Interaft in nort in nort in nort in nort in nort in nort in nort in norts in nort in nort in norts in nort in norts in nort in nort in norts cr中。同种异体移植物，表明某些CR是由Th2细胞介导的。

项目成果

Silver J, Gulwani-Akolkar B, Leffell MS, Kotb M, Obata F, Kashiwagi N.: "HLA and the T cell receptor."Charoone D (ed) Genetic diversity of HLA : Functional and Medical Implications. Vol I.EDK, Paris. 183-187 (1997)

Silver J、Gulwani-Akolkar B、Leffell MS、Kotb M、Obata F、Kashiwagi N.：“HLA 和 T 细胞受体”。Charoone D（编辑）HLA 的遗传多样性：功能和医学影响。

Ito K, Tanaka T, Tsutsumi N, Obata F, Kashiwagi N.: "Possible mechanisms of immunotherapy for maintaining pregnancy in recurrent spontaneous aborters : Analysis of anti-idiotypic antibodies directed against autologous T-cell receptors"Hum Reprod. 14-3. (1

Ito K、Tanaka T、Ttsutsumi N、Obata F、Kashiwagi N.：“复发性自发流产者维持妊娠的免疫疗法的可能机制：针对自体 T 细胞受体的抗独特型抗体的分析”Hum Reprod。

Ito K et al.: "Possible mechanisms of immunotherapy for maintaining pregnancy in recurrent spontaneous aborters:Analysis of anti-idiotypic ant"Human Reproduction. 14. 650-655 (1999)

Ito K 等人：“免疫疗法维持复发性自发流产者妊娠的可能机制：抗独特型蚂蚁的分析”人类生殖。

Silver J et al.: "Genetic diversity of HLA : Functional and Medical Implications,Vol I." EDK, 5 (1997)

Silver J 等人：“HLA 的遗传多样性：功能和医学影响，第一卷”。

Obata F,Kumano K,Endo T,Takeuchi Y,Ohkubo M,Tsunoda-Iizuka M: "T-cell receptor variable gene analysis of renal allograft infiltrating cells in biopsy specimens using a non radioisotopic micromethod."Transplantation. 66・11. 1389-1392 (1998)

Obata F、Kumano K、Endo T、Takeuchi Y、Ohkubo M、Tsunoda-Iizuka M：“使用非放射性同位素显微方法对活检标本中的肾同种异体移植物浸润细胞进行 T 细胞受体可变基因分析。”移植。 -1392 (1998)