Research for HLA-DNA typing methods and its clinical aplication

1. HLA-DNA分型的发展首先，我们制作了许多引物用于HLA-DNA分型的PCR-SSOP方法。本方法可准确、准确地分型纯合子细胞组及肾移植供体和受体的HLA 2类抗原（HLA- dr，DP.DQ）。但SSOP方法耗时长，步骤繁琐，难以应用于临床。PCR-SSCP是一种简便易行的DAN-HLA分型新方法。通过对单链型、异双链型和双站立型的电泳分析进行修改，成功地将PCR-SSCP应用于HLA 2型分型。另一种简单、快速的HLA-DNA分型方法是PCR-SSP与DNA构象多态性分析相结合。这种方法将是最适合移植配型的策略之一。HLA- 2类抗原相容性与肾移植存活的关系采用上述PCR-DNA分型方法回顾性检测HLA- 1 -单倍相同肾移植中HLA- 2类抗原。令人惊讶的是，属于DQB1零错配组和DRB1一错配组的9名患者中有7名出现了早期移植物丢失。因此，该组2年移植物存活率极低（48%），低于DQB1和DEB1 1不匹配组（100%）。我们推测，当供体的HLA-DQ抗原不能被受体识别时，抑制系统不能很好地发挥作用。这是关于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.