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

为了表征参与肾移植排斥反应的t细胞群，我使用活检标本进行了几次分析。研究结果如下：1)为了使用活检标本准确分析t细胞受体（TCR）多样性，我开发了一种非放射性同位素微法，其中通过锚定聚合酶链反应扩增的TCR cdna与固定在尼龙膜上的一系列TCR可变基因DNA杂交。2) TCR分析使我能够评估参与肾移植排斥反应的t细胞的克隆性。该方法特别适用于监测免疫抑制治疗排斥反应后克隆性的变化。3)肾浸润t细胞的克隆程度在急性排斥反应（AR）和慢性排斥反应（CR）中无显著差异。然而，AR组浸润细胞与外周血细胞的克隆性差异大于CR组，这表明在接受AR的同种异体移植物中积累了特定的t细胞1克隆。4)病毒感染的同种异体移植物通常表现出与拒绝的同种异体移植物相似的t细胞克隆性，仅通过克隆性分析难以区分。5) CR异体移植物中α - β T细胞、cd4阳性T细胞和cd8阳性T细胞的浸润程度与AR异体移植物无明显差异，表明CR作为AR是由这些免疫细胞介导的。6)同种异体移植中干扰素γ的检测频率在CR和AR之间无显著差异，表明Th1细胞参与了CR和AR。7)白细胞介素4仅在1 / 3的CR异体移植中检测到，而在AR异体移植中未检测到，提示部分CR是由Th2细胞介导的。

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.