重型β-地中海贫血患儿异基因造血干细胞移植术后淋巴细胞亚群重建分析

Monitoring lymphocyte subsets after β-mediterranean anemia allogeneic hematopoetic stem cell transplantation in children

目的动态监测重型β-地中海贫血患儿异基因造血干细胞移植(allo-HSCT)术前及术后第1、2、3、6和12个月淋巴细胞亚群比例的变化,探讨淋巴细胞的恢复情况及重建特点。方法选取行allo-HSCT的27例重型β-地中海贫血患儿,其中人类白细胞抗原(HLA)全相合移植术20例,HLA不全相合移植术7例。采用流式细胞术(FCM)动态监测患者移植前及术后1年内CD3^+T细胞、CD3+CD4+T细胞、CD3^+CD8^+T细胞、CD4^+/CD8^+比值、CD19^+B细胞及CD16^+CD56^+自然杀伤(NK)细胞比例的变化。结果与allo-HSCT术前比较,重型β-地中海贫血患儿术后CD16^+CD56^+NK细胞比例恢复最快,术后第1、2、3、6个月均高于术前(P<0.05),第12个月恢复至术前水平(P>0.05)。CD3^+T细胞比例术后第1个月与术前比较差异无统计学意义(P>0.05),第2个月起降低且低于术前(P<0.01),第12个月仍低于术前(P<0.01)。CD3^+CD8^+T细胞比例术后第1个月明显高于术前(P<0.01),随后逐渐下降,至术后第12个月时仍高于术前(P<0.05)。CD3^+CD4^+T细胞比例术后明显低于术前(P<0.01),且呈缓慢回升趋势,至术后第12个月仍明显低于术前(P<0.01)。CD4^+/CD8^+比值长期倒置,虽然呈缓慢上升趋势,但仍明显低于术前(P<0.01)。CD19^+B细胞比例术后第1个月明显低于术前(P<0.01),术后第2个月恢复至术前水平(P>0.05),术后第6个月和第12个月明显高于术前(P<0.01、P<0.05)。术前及术后同一时间点男性、女性患儿之间及HLA全相合移植与HLA不全相合移植之间各淋巴细胞亚群比例差异均无统计学意义(P>0.05)。结论重型β-地中海贫血患儿allo-HSCT术后淋巴细胞亚群的免疫重建恢复顺序依次为CD16^+CD56^+NK细胞、CD3^+CD8^+T细胞、CD19^+B细胞、CD3^+T细胞、CD3^+CD4^+T细胞。

Objective To monitor the development of lymphocyte subsets on the 1st,2nd,3rd,6th and 12th months dynamically after allogeneic hematopoetic stem cell transplantation(allo-HSCT)in children with β-mediterranean anemia,and to observe the immune reconstitution and characteristic of lymphocyte subsets.Methods A total of 27 patients with severe β-mediterranean anemia who underwent allo-HSCT were enrolled.Among them,20 cases received human leukocyte antigen(HLA)matched transplantation,and 7 cases received HLA mismatched transplantation.By flow cytometry,the percentage changes of CD3^+ T cells,CD3^+CD4^+ T cells,CD3^+CD8^+ T cells,CD4^+/CD8^+ ratio,CD19^+ B cells and CD16^+CD56^+ natural killer(NK)cells were monitored dynamically in 1 year.Results CD16^+CD56^+ NK cells showed the fastest recovery,the percentages on the 1st,2nd,3rd and 6th months were higher than those of pre-operation(P<0.05),and it returned to the percentages of pre-operation on the 12th month(P>0.05).The percentage of CD3^+ T cells had no statistical significance on the 1st month compared to pre-operation(P>0.05),but it started to decrease on the 2nd month(P<0.01),and on the 12th month it was still lower than that of pre-operation(P<0.01).The percentage of CD3^+CD8^+ T cells on the 1st month was higher than that of pre-operation(P<0.01),and then it started to decrease but still high on the 12th month(P<0.05).The percentage of CD3^+CD4^+ T cells was lower than that of pre-operation(P<0.01),showing a trend of slow recovery,and was still lower than that of pre-operation,even on the 12th month(P<0.01).CD4^+/CD8^+ ratio inverted in the long term,but it was still lower than that of pre-operation although it continued to show increase(P<0.01).CD19^+ B cells decreased on the 1st month(P<0.01)then recovered after the 2nd month(P>0.05),which were higher than those of pre-operation at the 6th and 12th months(P<0.01,P<0.05).The immune reconstitution had no statistical significance at each time point between male and female patients(P>0.05),and so did HLA matched transplantation and mismatched transplantation(P>0.05).Conclusions The recovery of lymphocyte subsets afterβ-mediterranean anemia allo-HSCT in children following the regulation:CD16^+CD56^+ ^+ B cells,CD3^+ T cells and CD3^+CD4^+ T cells.