Research for HLA-DNA typing methods and its clinical aplication

1. IILA-DNA分型方法的建立首先，我们研制了多种引物，用于HLA-DNA分型的PCR-SSOP方法。应用该方法可对同种异体分型细胞组及肾移植供、受者的HLA Ⅱ类抗原（HLA-DR、DP、DQ）进行准确、准确的分型。本文介绍了一种新的简便易行的DAN-HLA分型方法--PCR-SSCP，通过对电泳图谱分析方法的改进，成功地将PCR-SSCP应用于HLA 2分型（单链形式，HLA-DNA分型方法的建立是一种简便、快速的方法。SSP结合DNA构象多态性分析。该方法将是最适合移植配型的策略之一. HLA Ⅱ类抗原与移植肾存活的关系应用上述PCR-DNA分型方法对HLA单倍相合的肾移植患者进行了HLA Ⅱ类抗原的回顾性研究，发现DQB 1 0配型和DRB 1 1配型的9例患者中，有7例发生了早期移植肾失活。因此，这一组的2年移植物存活率（48%）明显低于DQB 1和DEB 1单错配组（100%），我们推测，当供体的HLA-DQ抗原不被受体识别时，抑制系统不能很好地发挥作用。这是抑制基因在临床移植中的首次报道。

1. Development of IILA-DNA typingAt first, we made many primers for PCR-SSOP method of HLA-DNA typing. With this method we could type HLA class 2 antigens (HLA-DR,DP.DQ) of homozygousu typing cell panels and donor and recipients of kidney transplantation, precisely and correctly. But application of SSOP method for clinical use is difficult by its time consuming and conjugated steps.As a new simple and easy DAN-HLA typing method we introduced PCR-SSCP.Application of PCR-SSCP for HLA class 2 typing was successful by the modification of analysis of electrophoretic pattern (single-stranded form, heteroduplex form and double-standed form).Another a simple and rapid HLA-DNA typing method was developed using PCR-SSP in combination with DNA conformation polymorphism analysis. This method will be one of the most suitable strategies for transplantaion matching.2. Correlation between HLA class 2 antigens compatibility and survial of kidney transplantationIn HLA-one-haplo-identical renal transplantation HLA class 2 antigens was examined retrospectively by the PCR-DNA typing methods which mentioned above.Surprisingly, the seven of nine patients who belonged to DQB1 zero-mismatched and DRB1 one-mismatched group had early graft loss. Accordingly, the 2 year graft survival in this group was extremely lower (48%) than that in the DQB1 and DEB1 one mismatch group (100%) .We speculate that the suppressor system done not function well when the donor's HLA-DQ antigens is not recognized by the recipient. This is a first report about suppression gene in clinical transplantation.