异基因造血干细胞移植后免疫重建的临床研究及急性移植物抗宿主病的危险因素分析

摘要

Objective To investigate the relationship between the reconstitution of lymphocyte subsets after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and acute graft-versus-host disease (aGVHD),and to explore the risk factors of aGVHD.Methods The proportions of peripheral blood lymphocyte subsets of 65 patients undergoing allo-HSCT during Jan 2012 and Nov 2013 were retrospectively analyzed.The lymphocyte subsets proportion in peripheral blood lymphocyte was studied by flow cytometry on 1st,3rd,6th and 12nd month after transplantation,respectively.The risk factors of aGVHD were analyzed by Logistic regression.Results The proportion of CD3+ T subsets of the peripheral blood lymphocyte in patients who accepted allo-HSCT was decreased obviously since implantation.The proportion of CD3+ T subsets on 1st month after transplantation was decreased significantly,then gradually rose to normal [(63.65±6.64) ％] on 3rd month after transplantation.Among them,the proportion of CD3+ CD4+ T subsets was lowest on 1st month after transplantation and was reconstructed slowly to normal until 12nd month after transplantation [(23.94 ± 3.15) ％].Proportion of CD3+ CD8+ T subsets was decreased and rose to normal [(31.06±8.40)％] on 1st month after transplantation.CD3+ CD4+ and CD3+ CD8+ ratio was at a low level after transplantation,and still lower than normal at 12nd month after a slow recovery,which was approximately 0.54±0.11.The proportion of CD3+ CD25+ was low on 1st month [(2.16±0.93)％] and was increased to normal at 3rd month.Proportion of CD3-CD16+ CD56+ cells was increased significantly to (42.31±8.47) ％,and gradually fallen down to normal at 6th month.In the first 3 months after implantation,CD3+ T cells from patients with aGVHD (grade Ⅲ-Ⅳ) significantly were increased comparing with patients without aGVHD (1 month:P =0.037,3 months:P =0.045) as well as CD3+ CD25+ T cells (1 month:P =0.025,3 months:P =0.041),which was opposite to CD3+ CD4+/CD3+ CD8+ (1 month:P =0.040,3 months:P =0.043).On 1st month after transplantation,CD3+ CD4+ T cells from patients without aGVHD displayed higher than that from patients with aGVHD (grade Ⅲ-Ⅳ) (P =0.032) as well as CD3-CD16+ CD56+ cells (P =0.046),which was opposite to CD3+ CD8+ cells (P =0.029).The results of multivariate analysis showed that donor-recipient HLA mismatching (OR =2.511,P =0.024),unrelated donor (OR =2.964,P =0.018) and GVHD prophylaxis without ATG (OR =2.792,P =0.033) were the risk factors for aGVHD.Conclusion Our results support the utility of post-transplant monitoring of a peripheral blood lymphocyte subset for early warning aGVHD of patients undergoing allo-HSCT.Donorrecipient HLA mismatching,unrelated donor and GVHD prophylaxis without ATG are the risk factors for aGVHD.%目的 探讨异基因造血干细胞移植后患者外周血淋巴细胞亚群表达水平与急性移植物抗宿主病(aGVHD)的相关性,探究移植后免疫重建的一般规律及引起aGVHD的危险因素.方法 回顾性分析2012年1月到2013年11月行异基因造血干细胞移植的65例患者临床资料,应用流式细胞术检测患者移植后1、3、6、12个月外周血淋巴细胞亚群表达水平,研究免疫重建规律,分析淋巴细胞亚群与aGVHD的相关性;运用Logistic回归分析aGVHD的危险因素.结果 移植后1个月CD3+比例下降,移植后3个月回升至正常,平均为(63.65±6.64)％;其中,CD3+ CD4+比例在移植后降低,移植后12个月恢复至正常,达(23.94±3.15)％;而CD3+ CD8+比例恢复较快,移植后1个月即接近正常,达(31.06±8.40)％;CD3+ CD4+/CD3+ CD8+比值在移植后处于低水平,后缓慢回升,但移植后12个月时仍处于倒置,为0.54+0.11.CD3+ CD25+比例在移植后1个月低于正常水平,为(2.16±0.93)％,3个月恢复正常;CD3-CD15+ CD56+比例在移植后1个月明显升高,达(42.31±8.47)％,移植后6个月逐渐回落至正常.移植后的前3个月,Ⅲ～Ⅳ度aGVHD组的CD3+比例高于无aGVHD组(1个月时,P=0.037;3个月时,P=0.045),CD3+ CD25+比例高于无aGVHD组(1个月时,P=0.025;3个月时,P=0.041),CD3+ CD4+/CD3+ CD8+比值低于无aGVHD组(1个月时,P=0.040;3个月时,P=0.043).移植后1个月无aGVHD组的CD3+ CD4+比例高于Ⅲ～Ⅳ度aGVHD组(P=0.032),CD3-CD15+ CD56+比例高于Ⅲ～Ⅳ度aGVHD组(P=0.046),CD3+ CD8+比例低于Ⅲ～Ⅳ度aGVHD组(P=0.029).供受体HLA配型不全合(OR =2.511,P=0.024)、无关供体(OR=2.964,P=0.018)、不使用抗胸腺细胞球蛋白(ATG)预防GVHD(OR=2.792,P=0.033)是发生aGVHD的危险因素.结论 移植后早期的T细胞免疫重建主要以非胸腺依赖性的CD8+细胞为主,而胸腺功能的损伤可引起依赖胸腺途径再生的T细胞(如CD4+细胞)重建迟缓.CD25+T细胞与aGVHD有密切关联,其表达水平与aGVHD的严重程度呈正相关.供受体HLA配型不全合、无关供体、不使用ATG预防是发生aGVHD的危险因素.