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深圳市健康无偿献血者外周血淋巴细胞表面人类白细胞抗原-E的基因型特异性表达研究

Detection of genotype-specific expressions of human leukocyte antigen-E on surface of peripheral blood lymphocytes of voluntary blood donors in Shenzhen
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摘要 目的探讨深圳市无偿献血个体的外周血淋巴细胞表面人类白细胞抗原(HLA)-E表达水平,并分析HLA-E基因型与HLA表达水平的相关性。方法采用简单随机抽样法选择2015年8月至10月于深圳市血液中心无偿献血的82例献血者作为研究对象。采用Sepmate淋巴细胞分离管分离82例献血者的外周血淋巴细胞,然后采用流式细胞技术检测淋巴细胞表面HLA-E的表达水平;同时提取研究对象的外周血的DNA,采用PCR-直接测序法(SBT)进行序列测定,判定其HLA-E基因型;并且采用方差分析和t检验的统计学方法,分析比较不同HLA-E基因型献血者外周血淋巴细胞表面HLA-E表达水平的差异。结果本研究82例献血者中,检出的HLA-E*01:01/*01:01基因型频率为20.7%(17/82),HLA-E*01:Ol/*01:03基因型频率为41.5%(34/82),HLA-E*01:03/*01:03基因型频率为37.8%(31/82),3种基因型献血者的外周血淋巴细胞表面HLA_E相对表达水平分别为2.57±0.66、3.32±1.33和3.77±1.26,三者比较,差异有统计学意义(F=6.062,P〈0.05)。其中,HLA-E*01:03/*01:03基因型献血者外周血淋巴细胞表面HLA-E相对表达量最高,其次为HLA-E*01:01/*01:03基因型献血者,HLA-E*01:0l/*01:01基因型献血者外周血淋巴细胞表面HI。A-E相对表达量最低,并且两两比较,差异均有统计学意义(HLA-E;01:03/*01:03基因型与HI。A_E*01:01/*01:03基因型比较,t=2.402,P〈0.05;HLA-E*01:03/*01:03基因型与HLA-E*01:01/*01:01基因型比较,t=4.212,P〈0.001;HLA-E*01:Ol/*01:03基因型与HLA-E*01:01/*01:01基因型比较,t=3.054,P〈0.01)。结论深圳市健康献血人群外周血淋巴细胞表面HLA-E表达水平与其基因型密切相关。 Objective To detect the expression level of human leukocyte antigen (HLA)-E on surface of human peripheral blood lymphocytes in Shenzhen voluntary blood donors, and analyze the correlation between the genotype of HLA-E and the expression level of HLA-E. Methods From August to October 2015, a total of 82 blood donors were selected as the research object by simple random sampling method in Shenzhen Blood Center. Lymphocytes were separated from human peripheral blood by Sepmate lymphocyte separation tubes in these 82 blood donors, and then the expression level of HLA-E on the surface of lymphocyte was detected by flow cytometry. DNA of all donors were extracted from the peripheral blood samples to identify the genotype of HLA-E by PCR-sequence-based typing (SBT) method. Statistical methods were used to analyze the correlation between the expression level of HLA-E on surface of peripheral blood lymphocytes and its HLA-E genotype. Results The genotype frequency of HLA-E * 01 : 01/* 01 : 01 was 20. 7% (17/82), that of HLA-E * 01 : 01/01 : 03 was 41.5% (34/82), and that of HLA-E * 01 : 03/* 01:03 was 37.80/oo (31/82). The relative expression levels of HLA-E on surface of peripheral blood lymphocytes of different genotype donors were 2.57±0.66, 3. 32±1.33 and 3.77±1.26, respectively, and the difference was statistically significant (F=6. 062, P〈0.05). The highest HLA-E expression level was in HLA-E * 01 : 03/*01 : 03 genotype blood donorsr peripheral blood lymphocytes surface, followed by HLA-E *01 : 01/* 01 : 03 genotype, and the lowest was HLA-E* 01 : 01/* 01 : 01 genotype, while the differences of multiple comparison were statistically significant (HLA-E * 01 : 03/* 01 : 03 genotype vs HLA-E* 01 : 01/* 01 : 03 genotype: t=2. 402, P(0. 05; HLA-E* 01 : 03/* 01 : 03 genotype vs HLA-E* 01 : 01/*01 : 01 genotype: t=4. 212, P〈0. 001; HLA-E * 01 : 01/* 01 : 03 genotype vs HLA-E * 01 : 01/* 01 :
出处 《国际输血及血液学杂志》 CAS 2017年第2期137-143,共7页 International Journal of Blood Transfusion and Hematology
基金 广东省医学科研基金(A2016222) 深圳市战略新兴产业发展专项资金(JSGG20160328103642937)
关键词 淋巴细胞 HLA抗原 基因表达 基因型 Lymphocytes HLA antigens Gene expression Genotype
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