人外周血NK细胞亚群、表型和生物学特征

Phenotypic and biological characterization of human NK cells in PBMCs

目的:探讨人外周血NK细胞的异质性和生物学特征。方法:利用多种标记抗体进行细胞表面和细胞内细胞毒效应分子染色,再利用流式细胞仪在单个细胞水平上分析NK细胞亚群的多样性和生物学特征。结果:NK细胞可分为CD56+、CD56+CD16+和CD16+三个亚群。所有CD56+NK细胞均表达CD95,但表达水平的高(CD95bright)和低(CD95dim)不同。在CD95bright和CD8+细胞亚群中有43.5%的细胞同时表达颗粒酶B和穿孔素。而CD56+CD16+和CD16+细胞表达CD95均为CD95bright,同时表达颗粒酶B(83.6%)和穿孔素(89.8%)。结论:NK细胞乃异质性群体,随着CD56表达减少和CD16表达增加,其生物效应功能逐渐成熟。

Objective:To characterize the phenotypic and biological properties of NK cells in human PBMCs.Methods:PBMCs were isolated from normal individuals. Surface markers and intracellular cytotoxic molecules of PBMCs were stained with multi-color-labeled monoclonal antibodies and analyzed at the single cell level the relation between NK subsets and biological characterization by flow cytometer.Results:Three distinct subpopulations(CD56+,CD56+CD16+ and CD16+) of human NK cells could be identified based upon the expression of CD56 and CD16 molecules. All of CD56+ NK cells expressed CD95 but some of them revealed CD95 bright and CD95 dim.5%-6% of CD56+NK cells were CD8+;43.5% of CD95 bright and CD8+ subsets expressed Granzyme B and Perforin,whereas 1%-6% of other subsets were positive for Granzyme B and Perforin. CD56+CD16+ and CD16+ subsets were CD95 bright,whereas 32.2% of them were CD8+.Both CD56+CD16+ and CD16+ subsets expressed Granzyme B and Perforin were 83.6% and 89.8%,respectively,but 7.35% of CD56+CD16+ subset was negative for both Granzyme B and Perforin.Conclusion:NK cells in PBMCs are phenotypically and biologically heterogenous.With the reduction in the expression of CD56 and the increase in the expression of CD16 molecules,NK cells are gradually becoming maturation of biological properties.