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

为了确定参与肾移植排斥反应的T细胞群体的特征，我使用活组织标本进行了几项分析。结果如下：1)为了准确分析活检标本中T细胞受体(TCR)的多样性，我建立了一种非放射性同位素微量方法，将锚定聚合酶链式反应扩增的TCR cDNA与固定在尼龙膜上的TCR可变基因DNA阵列杂交。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。3)急性排斥反应(AR)和慢性排斥反应(CR)肾组织浸润性T细胞的克隆性程度无明显差异。4)病毒感染的同种异体移植物通常表现出与排斥同种异体移植物相似的T细胞克隆性，仅通过克隆性分析很难区分它们。5)移植物中的α-βT细胞、CD4+T细胞和CD8+T细胞在CR移植物中的渗透程度与AR移植物中的无差异，表明CR作为AR的移植物，6)干扰素-γ在CR和AR中的表达频率无显著差异，提示Th1细胞同时参与CR和AR。7)IL-4仅在三分之一的CR移植物中检测到，而在AR移植物中未检测到，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.