HLA分子三维结构与GVHD相关性研究

Study on the relationship between HLA three-dimensional structure and GVHD in bone marrow transplantation

目的 进一步探索GVHD的发病机制，提高临床移植疗效。方法 HLA分型方法包括血清一细胞学、基因和基因亚型分型。血清一细胞学分型采用国际通用的微量淋巴细胞毒试验和混合淋巴细胞培养；基因分型采用PCR-SSP方法．对基因亚型进行DNA测序，再进行HLA三维结构模拟。结果 (1)供一受体间HLA-Ⅰ、Ⅱ类血清学相合，组基因相合，基因亚型不同．但HLA三维结构模拟两者差别不明显，骨髓移植后仅发生了Ⅱ度GVHD；(2)供一受体间，基因亚型不同，两者仅差1个氨基酸，供一受体间抗原三维结构差别明显，骨髓移植后发生IV度GVHD；(3)受体为HLA-A°0201，供体为A°6801，二者血清学呈强交叉反应，其它位点均相合，移植后发生IV度GVHD，后经HLA三维结构模拟，两者差别明显。结论 当异基因骨髓移植供一受体问HLA基因亚型不同时，移植后GVHD的发生程度与HLA分子三维结构差别大小有密切关系；GVHD与抗原氨基酸差别的数量没有直接关系。

Objective To investigate the mechnism and enhance the effect of BMT in clinic. Methods HLA typing methods included serology-cytology,genetic and subgenetic typing. Microlymphocytetotoxicity and mixed leucocyte culture assay were used in the studies ;PCR-SSP for genotyping was used;modeling of three-dimensional structure was performed after DNA sequence analysis. Results (l)HLA serotype and group genotype were matched between donor and recipient .HLA-DRB1' 04 was observed. HLA molecules had not distinct differences on modeling of three-dimensional structure between donor and recipient, I degree GVHD was developed after BMT ; (2)There was one difference on animo acid of HLA molecules between donor and recipient with different subgenotype of HLA.three-dimensional structure were distinct different,so IV degree GVHD was caused ; (3)Subgenotype HLA-A' 0201 was observed in patient and A'6801 in donor ,serotyping appeared distinct cross-reactions between donor and recipient ,resulted in IV degree GVHD after BMT. And distinct differences of three-dimensional structures were found. Conclusions When HLA subgenotypes were different in allogenic BMT,the stages of GVHD after BMT were closely associated with stages of differentiation of three-dimensional structure between donor and recipient; there was no direct relationship between GVHD and different number of animo acid.