摘要目的 探讨人类白细胞抗原(HLA)-E?01:03阳性基因型与急性白血病(AL)的相关性.方法 选择2013年1月至2015年12月,于深圳市血液中心进行血型配型的136例AL患者为研究对象,纳入AL组.其中,男性患者为78例,女性为58例;年龄为(39±23)岁.采用简单随机抽样法选择同期于深圳市血液中心参加无偿献血的182例健康献血者纳入对照组.其中,男性献血者为101例,女性为71例;年龄为(38±19)岁.采用PCR-直接测序分型(SBT)法对2组受试者的HLA-E基因第3外显子进行序列测定,并且判断HLA-E基因型.采用密度梯度离心法分离AL组54例患者和对照组64例健康献血者的外周血淋巴细胞;采用流式细胞术检测淋巴细胞表面HLA-E相对表达水平.采用酶联免疫吸附试验法(ELISA)检测AL组54例患者和对照组64例健康献血者的血浆可溶性HLA(sHLA)-E表达水平.2组受试者基因型频率比较采用χ2检验,淋巴细胞表面HLA-E相对表达水平、血浆中sHLA-E表达水平比较,采用两独立样本t检验.本研究遵循的程序符合本研究遵循的程序符合深圳市血液中心医学伦理委员会制定的标准,经过该伦理委员会批准(批准文号:SZBC-2017-007),并且与所有受试者签署临床研究知情同意书.结果 ①本研究AL组患者的HLA-E?01:03阳性基因型频率为90.4％(123/136),高于对照组健康献血者的77.5％(141/182),并且差异有统计学意义(χ2=9.286,P=0.002).②本研究AL组54例患者与对照组64例健康献血者中,2组HLA-E?01:03阳性基因型受试者的外周血淋巴细胞表面HLA-E相对表达水平比较,差异无统计学意义[(3.3±0.4)比(3.6±0.2);t=0.77,P=0.440].AL组HLA-E?01:03阴性基因型患者外周血淋巴细胞表面HLA-E相对表达水平为(1.6±0.2),低于对照组的(2.5±0.2),并且差异有统计学意义(t=2.95,P=0.010).③本研究AL组54例患者与对照组64例健康献血者中,AL组HLA-E?01:03阳性基因型患者的血浆sHLA-E表达水平为(31.2±0.4)pg/mL,高于对照组的(18.2±0.3)pg/mL,并且差异具有统计学意义(t=26.63,P<0.001);AL组HLA-E?01:03阴性基因型患者血浆sHLA-E表达水平为(32.9±1.3)pg/mL,亦低于对照组的(18.8±0.8)pg/mL,并且差异亦有统计学意义(t=8.89,P<0.001).结论 HLA-E?01:03阳性基因型频率及血浆sHLA-E表达水平与AL具有一定的相关性,其可能成为AL的潜在的预测及辅助诊断指标.

abstractsObjective To analyze relationship between positive genotype of human leukocyte antigen (HLA)-E?01:03 and acute leukemia (AL).Methods From January 2013 to December 2015,a total of 136 patients with AL who underwent blood group matching at Shenzhen Blood Center were included in this study as AL group.Among them,there were 78 male patients and 58 females;the age was (39±23)years. A simple random sampling method was used to select 182 health blood donors who participated in voluntary blood donation at the Shenzhen Blood Center in the same period.Among them,there were 101 male donors and 71 females;and age was (38±19)years.The sequence of exon 3 of HLA-E gene was determined by PCR-sequence based typing (SBT)method,and the genotype of HLA-E gene was determined.The peripheral blood lymphocytes from 54 patients in AL group and 64 healthy blood donors in control group were separated by density gradient centrifugation.The relative expressions of HLA-E on lymphocytes were detected by flow cytometry.The expression levels of plasma soluble HLA (sHLA)-E from 54 patients in AL group and 64 healthy blood donors in control group were detected by enzyme-linked immunosorbent assay (ELISA).The genotype frequencies of subj ects in the two groups were compared by Chi-square test. The relative expression levels of HLA-E on peripheral blood lymphocytes and the expression level of sHLA-E in plasma were compared by independent samples t test,respectively.This study was in line with the procedures followed in this study were in accordance with the standards established by the Medical Ethics Committee of the Shenzhen Blood Center,and this study was approved by the committee (Approval No.SZBC-2017-007).All the subjects signed the informed consents for clinical trials and informed contents were obtained from all subjects.Results ① The genotype frequency of HLA-E?01:03 positive genotype was 90.4％ (123/136)of patients in AL group,which was higher than that of 77.5％ (141/182)in control group,and the difference was statistically significant (χ2=9.286,P=0.002).② Among the 54 patients in AL group and the 64 healthy blood donors in control group,the difference of relative expression levels of HLA-E on peripheral blood lymphocytes from subjects with HLA-E?01:03 positive genotype between the two groups were not statistically significant [(3.3±0.4)vs (3.6±0.2);t=0.77,P=0.440].The relative expression level of HLA-E on peripheral blood lymphocytes from patients with HLA-E?01:03 negative genotype in AL group was (1.6±0.2),which was lower than that of (2.5 ± 0.2)in control group,and the difference was statistically significant (t=2.95,P=0.010).③ Among 54 patients in AL group and 64 healthy blood donors in control group,the expression level of sHLA-E in plasma from patients with HLA-E?01:03 positive genotype in AL group was (31.2±0.4 )pg/mL,which was higher than that of (18.2±0.3)pg/mL in control group,and the difference was statistically significant (t=26.63,P<0.001). The expression level of sHLA-E in plasma from patients with HLA-E?01:03 negative genotype in AL group was (32.9±1.3)pg/mL,which was lower than that of (18.8±0.8)pg/mL in the control group,and the difference was also statistically significant (t=8.89,P<0.001 ).Conclusions The HLA-E?01:03 positive genotype frequency and the expression of sHLA-E in plasma have a certain correlation with AL.They may be a potential predictor and an auxiliary diagnostic indicator for AL.