异基因造血干细胞移植受者外周血中可溶性HLA-G与急性GVHD的相关性

The correlation between soluble HLA-G and acute graft-versus-host disease after allo-geneic hematopoietie stem cell transplantation

目的 探讨异基因造血干细胞移植（allo-HSCT）受者外周血中可溶性HLA-G（sHLA-G）的水平与急性移植物抗宿主病（aGVHD）间的相关性以及与CD4＋CD25^（hlgh）调节性T淋巴细胞（Treg）水平的相关性.方法 选择27例allo-HSCT受者,将其中13例术后发生aGVHD者作为aGVHD组,14例未发生aGVHD者作为对照组.在移植预处理前、移植后第2、4、8、14周时,分别抽取两组受者清晨空腹肘静脉血2～4 ml,采用酶联免疫吸附试验双抗夹心法定量检测sHLA-G,采用流式细胞术检测CD4＋CD25＋Treg.动态监测术后第2和14周受者sHLA-G水平的变化;动态监测和分析术后第8和14周两组受者间sHLA-G的差异.观察sHLA-G与发生aGVHD的相关性;检测术后第14周两组受者CD4＋CD25＋Treg的数量,观察sHLA-G水平与CD4＋CD25＋Treg的相关性.结果 预处理前,对照组和aGVHD组受者sHLA-G的水平分别为（75.4±13.1）/μg/L和（47.0±14.1）btg/L,两组间差异有统计学意义（P〈0.05）.术后第8周和第14周,对照组受者sHLA-G的水平明显高于相应时间点aGVHD组受者,两组间差异均有统计学意义（P〈0.01）.对照组受者术后第14周的sHLA-G水平与第2周相比,增长了1.2～3.4倍,两时间点的差异有统计学意义（P〈0.05）.术后第14周,通过Spearman等级相关检验显示受者sHLA_G水平与CD4＋CD25＋Treg比例具有统计学上的相关性（相关系数r=0.810,P〈0.05）,结论 allo-HSCT后受者体内sHIA-G对aGVHD的发生起负性调节作用;sHLA-G的水平与外周血CD4＋CD25^（hlgh） T淋巴细胞的比例旱正相关,二者之间可能存在相互诱导和调节的信号途径,在诱导和维持移植免疫耐受中起着重要的协同作用.

Objective To explore the effect of sHLA-G on acute GVHD（aGVHD） and the relationship between the level of sHLA-G and the number of CD4＋ CD25^（hlgh）T cells （Treg） after allogeneic hematopoietic stem cell transplantation （allo-HSCT）.Methods Among 27 cases of allo-HSCT recipients, 13 of them with aGVHD served as aGVHD group, and the rest 14 as control group.Before and at the 2nd, 4th, 8th, 14th week after transplantation, 2-4 ml blood was obtained from elbow veins in the early morning, double-antibody sandwich of enzyme-linked immunosorbent assay was used for quantification of sHLA-G, and flow cytometry for dynamic determination of the number of CD4~＋ CD25~＋ Treg.The levels of sHLA-G at the 8th and 14th week after transplantation in the recipients were measured, and the relationship between the sHLA-G and the occurrence of aGVHD was analyzed.By detecting the number of CD4~＋ CD25~＋ Treg from two recipients groups at the 14th week, the relevance between the level of sHLA-G and the number of CD4＋ CD25＋ Treg was observed.Results Before transplantation, the levels of sHLA-G in the control group and aGVHD group were （75.4±13.1 ） and （47.0±14.1 ） μg/L respectively with the difference being statistically significant （P〈0.05）.At the 8th and 14th week after transplantation, the levels of sHLA-G in the control group were significantly higher than in the aGVHD group at the corresponding time points with the difference being statistically significant （P〈0.01 ）.In the control group, the levels of sHLA-G at the 14th week were increased by 1.2-3.4 times, compared with those at the 2nd week （P.〈 0.05）.At the 14th week, Spearman rank correlation tests revealed that there was a statistical correlation between the levels of sHLA-G from the recipients and the proportion of CD4＋ CD25＋ Treg （correlation coefficient was 0.810, P〈0.05）.Conclusion The plasma sHLA-G in patients after allo-HSCT plays a negative regulatory role in the occurrence of aGVHD.There is a positive correlation between the levels of sHLA-G and the proportion of CD4＋ CD25^（hlgh）Treg in peripheral blood.There may exist induction and regulation of molecules through the cell signaling pathways between sHLA-G and CD4＋ CD25^（hlgh） Treg, which can synergically induce and maintain transplant immune tolerance.