人类白细胞抗原G基因多态性与急性移植物抗宿主病的相关性研究

Study on Correlation of the HLA-G Polymorphism with Acute Graft-Versus-Host Disease After HLA-Identical Sibling Hematopoietic Stem Cell Transplantation

目的：通过研究人类白细胞抗原G（HLA-G）基因多态性与同胞全相合造血干细胞移植（HSCT）术后急性移植物抗宿主病（aGVHD）的关系，探讨HLA-G能否作为移植前评估aGVHD发生的指标。方法采集82例同胞全相合造血干细胞移植患者预处理-9 d、14 d、28 d外周血2 mL，予肝素抗凝。留取供体造血干细胞采集物2 mL，予肝素抗凝，提取单个核细胞。利用DNA抽提试剂盒抽提患者与供者基因组DNA，应用聚合酶链反应（PCR）扩增基因组DNA，同时应用PCR产物双向测序的方法分析HLA-G基因3＆#39;端非编码区14 bp插入与缺失（deletion or insertion, del or ins）多态性，分为del/del组、del/ins组、ins/ins组。采用实时定量PCR方法检测患者移植前后HLA-G mRNA相对表达量，比较不同基因型之间和aGVHD患者与non-aGVHD患者之间相对表达量差异。结果发现14 bp ins纯合子患者mRNA表达水平低于del/del和del/ins患者，但未发现HLA-G 14 bp ins纯合子患者发生aGVHD的风险高于14 bp del患者。结论该实验数据并未发现HLA-G基因多态性能够作为预测异基因造血干细胞移植后aGVHD发生风险的危险因素。

Objective To explore the correlation of HLA-G with acute graft-versus-host disease （aGVHD） after sibling hematopoietic bone marrow transplantation （HSCT） and whether HLA-G can be used as a pre-transplant assessment of aGVHD risk factors.Methods We Collected the peripheral blood at multiple time points （d-9, d14, d28） from 82 patients after sibling HSCT and donor hematopoietic stem cells material with heparin, extracting mononuclear cells. HLA-G 14 bp polymorphisms were analyzed by PCR ampliifcation of the entire coding region followed by direct bidirectional DNA sequencing, deifned as group del/del, group del/ins, group ins/ins. Real-time quantitative PCR was used to detect the relative expression level of HLA-G or between aGVHD and non-aGVHD patients. Clinical data were retrospectively analyzed in patients receiving sibling mismatched HSCT. To analysis the relationship among the different genotypes and aGVHD.Results Analysis showed that the mRNAexpression levels were signiifcantly lower than the del/del and del/ins group before and after transplantation. And there were not statistically signiifcant association between the 14 bp ins/ins genotype and a lower risk of aGVHD compared to the 14 bp ins/del or del/del genotypes.Conclusion Our data show that there is no apparent value of considering polymorphisms in the 3＇UTR of HLA-G for risk prediction of aGVHD after allogeneic HSCT.