淋巴细胞分离液对T细胞阳性花环和非T细胞阳性花环计数结果影响的研究

摘要

目的 探讨单克隆抗体SPA红细胞花环法(McAb-A-E法)检验T淋巴细胞亚群时,淋巴细胞分离液对阳性花环细胞计数结果以及非T淋巴细胞阳性花环计数结果的影响.方法 选择淋巴细胞分离液法分离的单个核细胞(PBMC)和不加任何物质的抗凝血用自然沉降法分离的混合细胞,用10倍量稀释液洗涤1次.两种细胞用McAb-A-E直接法进行样本对照,并作统计学处理.结果 对20例临床标本用两种不同方法制取单个核细胞悬液的实验对照研究,2组CD3+ (t=-4.197,P=0.000)、CD4+(t=-5.177,P=0.000)、CD8+(t=-4.240,P=0.000)、CD4+/ CD8+(t=-1.958,P=0.095)进行比较,CD3+、CD4+、CD8+ 3组结果差异有统计学意义,均为P＜0.001.CD4+/ CD8+ 结果比较,差异无统计学意义(P＞0.05);2组非T淋巴细胞阳性花环比较(t=-5.638,P=0.000),差异有统计学意义(P＜0.001).结论 淋巴细胞分离液分离的PBMC细胞,用10倍量的稀释液洗涤1次,细胞表面黏附的黏稠淋巴细胞分离液是无法全部除去的,其对实用McAb-A-E直接法实验的影响是使阳性花环淋巴细胞计数结果偏高,相对的阴性淋巴细胞计数结果偏低.%Objective To study the influence of Lymphocyte Isolation Liquid to T cell and non-T cell positive Rosette. Methods To select the peripheral blood mononuclear cell(PBMC) separate from lymphocyte isolation liquid,and the natural separate hybrid cells without anticoagulation. The two kind＇s cell were washed once with 10 times liquefier of washing one time. The two cells with practical McAb A E. and the results were analyzed. Results From 20 cases sample, two kinds separate methods the results of CD3+ ,CD4+ ,CD8+ 3 results show statistic change(P＜0. 001) ,and the CD4+/CD8+ showed no statistic change(P＞0. 05). non-T cell Rosette Formation results in two group showed statistic change(t =- 5. 638, P = 0. 000). Conclusion The methods of using lymphocyte isolation liquid separate PBMC cell,then washes with one time with 10 time cannot separate the cell surface lymphocyte separating medium. And this will cause lymphocyte positive rosette counting result to be high, the relative negative rosette lymphocyte counting result is low.