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细胞群体，我使用活检标本进行了几项分析。结果表明：（1）为了准确分析肾移植患者T细胞受体（TCR）的多样性，作者建立了一种非放射性同位素的微量分析方法，将锚定聚合酶链反应扩增的TCR cDNA与固定在尼龙膜上的TCR可变基因DNA阵列进行杂交。3）急性排斥反应（AR）与慢性排斥反应（CR）肾组织浸润性T细胞克隆性程度无显著性差异。4）病毒感染的同种异体移植物常表现出与排斥移植物相似的T细胞克隆性，仅通过克隆性分析难以区分; 5）α-β T细胞的浸润程度，CR和AR移植物中的CD 4阳性T细胞、CD 8阳性T细胞无明显差异，表明CR与AR一样，是由这些免疫细胞介导的。6）CR和AR移植物中干扰素-γ的检测频率无明显差异，表明Th 1细胞参与了CR和AR。7）IL-4仅在三分之一的CR同种异体移植物中检测到，而在AR同种异体移植物中未检测到，表明一些CR由Th 2细胞介导。

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.