以流式细胞术建立中国汉族健康儿童外周血淋巴细胞亚群相对计数的正常参考值

The establishment of reference ranges of peripheral blood lymphocyte subpopulations for healthy Chinese Han children based on flow cytometry

目的建立中国汉族健康儿童外周血淋巴细胞亚群的正常参考值范围。方法选取首都医科大学附属北京儿童医院入托、入学体检,或术前查体及术后复查(均为对免疫功能影响不大的疾病)的0～18岁汉族儿童为研究对象。按年龄分为婴儿组(28d至12个月),幼儿组(～3岁),学龄前组(～7岁),学龄组(～12岁)和青春期组(～18岁)。采集外周血以双色及四色荧光标记流式细胞术检测淋巴细胞亚群T细胞(CD3+CD19-)、CD4+T细胞(CD3+CD4+)、CD8+T细胞(CD3+CD8+)、B细胞(CD3-CD19+)和NK细胞(CD3-CD16+CD56+)相对计数及CD4+/CD8+比值。比较各年龄组不同性别淋巴细胞亚群分布的差异,建立正常参考值范围。结果由于青春期组所收集的标本数较少,将该研究对象组予以删除。最终纳入28d至12岁儿童592例。①婴儿组T细胞、CD8+T细胞百分比与CD4+/CD8+比值性别差异均有统计学意义,幼儿组CD4+T细胞、CD8+T细胞百分比与CD4+/CD8+比值性别差异均有统计学意义,学龄组CD4+T细胞百分比性别差异有统计学意义;②除了男童T细胞百分比各年龄组间差异无统计学意义外,男女儿童各年龄组间外周各血淋巴细胞亚群分布总体上差异均有统计学意义;进一步行男女儿童淋巴细胞亚群不同年龄组间两两比较,发现多个年龄组间差异有统计学意义;③男女儿童T细胞、CD8+T细胞和NK细胞百分比随年龄增长均呈逐渐升高趋势;CD4+T细胞、B细胞百分比及CD4+/CD8+比值随年龄增长均呈逐渐降低趋势;男女儿童T细胞、B细胞、NK细胞百分比和CD4+/CD8+比值升高或降低的程度略有不同;④中国汉族健康儿童淋巴细胞亚群分布特点与欧、美、非洲国家儿童相比存在相似的升高或降低的趋势,但数值上存在一定的差异。结论儿童淋巴细胞亚群分布存在年龄和性别的差异。本研究成功建立中国汉族28d至12岁健康儿童淋巴细胞亚群相对计数正常参考值范围。

Objective To establish the reference ranges of peripheral blood lymphocyte subpopulations（T,B and NK cells）values in healthy Chinese children.Methods Chinese Han children who received admission,preoperative or postoperative physical examination aged of 0-18 years old were enrolled into the study.Included children were divided into infant group（28 d-12 months）,toddler age group（1-3 years）,preschool age group（3-7 years）,school age group（7-12-years）and adolescence age group（12-18 years）.Relative counts of lymphocyte subpopulations of T cell（CD3＋CD19-）,CD4＋T cell（CD3＋CD4＋）、CD8＋T cell（CD3＋CD8＋）、CD4＋/CD8＋、B cell（CD3-CD19＋）and NK cell（CD3-CD16＋CD56＋）were detected by flow cytometrics dual-color and four-color analysis.Differences in the percentages of lymphocyte subpopulations was compared between gender and among age groups,and the reference ranges of peripheral blood lymphocyte subpopulations were established.Results The adolescence age group was finally deleted becasue of the insufficient sample size.Then the percentages of lymphocyte subpopulations were determined in 592 healthy Chinese Han children aged of 28 d-12 years.①The interior-group comparison between males and females：the interior-group comparison showed significant differences between males and females in T,CD4＋T,CD8＋T lymphocyte percentages and CD4＋/CD8＋ value in infant group,toddler age group and school age group（P0.05）;②The inter-group comparison between all the lymphocyte subpopulations：the inter-group comparison showed significant differences between all the lymphocyte subpopulations（P0.01）except the T lymphocytes in males（P=0.602）,then the result of multiple comparison showed that there were statistical significances between several groups,especially between two younger than 3 years groups and two elder than 3 years of age groups（P0.01）,respectively;③The trend of peripheral blood lymphocyte subpopulations with age：the line chart showed T,CD8＋T and NK cell percentages were increased with age,CD4＋T,B cell percentages and CD4＋/CD8＋ values were declined with age;the line chart showed a little different trend in increasing or decreasing degree between males and females;④There was a similar trend in the percentages of lymphocyte subpopulations in different ethnic or regional groups such as Chinese,European,American and African,but a little different in numerical value.Conclusions The peripheral blood lymphocyte subpopulations values differ by gender and different age groups.The reference ranges of peripheral blood lymphocyte subpopulations values has been proposed by the current study for healthy Chinese Han children aged 28 d-12 years.