Hu X, He J, Zhang H H, Bao X J, Wang M, Zhang J, Cen J N, Wu X J, Yang X J
Jiangsu Institute of Hematology, Center for Clinical Laboratory, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Zhonghua Xue Ye Xue Za Zhi. 2017 Aug 14;38(8):667-672. doi: 10.3760/cma.j.issn.0253-2727.2017.08.004.
To investigate the immune reconstruct regularity profile of KIR2DL1 and KIR3DL1 in unrelated-donor allogeneic hematopoietic stem cell transplantation (allo-HSCT) with KIR-AA genotype. 75 donor-recipient pairs were performed by KIR genotying using PCR-SSP, and all donors were identified with KIR-AA genotype. Dynamic detections (including unrelated-donor on the day of transplantation and the recipient each month post allo-HSCT) of the expression of KIR2DL1/3DL1 on NK cell and mRNA level were performed in 291 cases using flow cytometry (FCM) and real-time fluorescent quantitation PCR (RT-qPCR) . ①The median expression of KIR2DL1 in unrelated-donor on transplant's day was 21.60%, the median expression of KIR2DL1 in recipient 1M, 2M, 3M and 3-6M after transplantation were 7.40%, 12.00%, 16.92%, 17.64% respectively. The median expression of KIR2DL1 in unrelated-donor on transplant's day was 265.14 copies/10 000abl copies, the median expression of KIR2DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 332.17, 438.31, 723.25, 414.17, 180.76 and 234.67 copies/10 000abl copies respectively. The median expression of KIR2DL1 on NK cells and mRNA level gradually increased at all time points after transplantation, and reached the highest expression at 3 months after transplantation. But mRNA expression levels increased earlier than NK cell membrane proteins. ②The median expression of KIR3DL1 in unrelated-donors on transplant's day was 18.56%, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M after transplantation were 23.83%, 22.57%, 23.02%, 21.60% respectively. The median expression of KIR3DL1 in unrelated-donor on transplant's day was 572.29 copies/10 000abl copies, the median expression of KIR3DL1 in recipient 1M, 2M, 3M, 3-6M, 6-9M, 9-12M after transplantation were 1 233.74, 1 140.42, 876.73, 1 057.07, 739.02 and 514.43 copies/10 000abl copies respectively. The median expression of KIR3DL1 on NK cells and mRNA level were higher than donors at 1 month after transplantation, and stable expression at all time points after transplantation, so mRNA and NK cell membrane proteins expression increased at the same time. The immune reconstruct regularity of KIR2DL1 and KIR3DL1 gene were different, which provided an experimental basis for selecting the best time to detect the expressions of KIR2DL1 and 3DL1 after transplantation.
探讨KIR-AA基因型非血缘供者异基因造血干细胞移植（allo-HSCT）中KIR2DL1和KIR3DL1的免疫重建规律。采用聚合酶链反应-序列特异性引物（PCR-SSP）对75对供受者进行KIR基因分型，所有供者均为KIR-AA基因型。采用流式细胞术（FCM）和实时荧光定量聚合酶链反应（RT-qPCR）对291例患者动态检测（包括移植当天非血缘供者及allo-HSCT术后受者每月1次）NK细胞表面KIR2DL1/3DL1的表达及mRNA水平。①移植当天非血缘供者KIR2DL1的表达中位数为21.60%，移植后1个月、2个月、3个月及3～6个月受者KIR2DL1的表达中位数分别为7.40%、12.00%、16.92%、17.64%。移植当天非血缘供者KIR2DL1的mRNA表达中位数为265.14拷贝/10 000个abl拷贝，移植后1个月、2个月、3个月、3～6个月、6～9个月、9～12个月受者KIR2DL1的mRNA表达中位数分别为332.17、438.31、723.25、414.17、180.76、234.67拷贝/10 000个abl拷贝。移植后各时间点NK细胞表面及mRNA水平KIR2DL1表达中位数逐渐升高，移植后3个月达最高表达，但mRNA表达水平升高早于NK细胞膜蛋白。②移植当天非血缘供者KIR3DL1的表达中位数为18.56%，移植后1个月、2个月、3个月、3～6个月受者KIR3DL1的表达中位数分别为23.83%、22.57%、23.02%、21.60%。移植当天非血缘供者KIR3DL1的mRNA表达中位数为572.29拷贝/10 000个abl拷贝，移植后1个月、2个月、3个月、3～6个月、6～9个月、9～12个月受者KIR3DL1的mRNA表达中位数分别为1 233.74、1 140.42、876.73、1 057.07、739.02、514.43拷贝/10 000个abl拷贝。移植后1个月受者NK细胞表面及mRNA水平KIR3DL1表达高于供者，移植后各时间点表达稳定，mRNA与NK细胞膜蛋白表达同步升高。KIR2DL1和KIR3DL1基因免疫重建规律不同，为移植后选择检测KIR2DL1和3DL1表达的最佳时间提供了实验依据。
