目的:通过研究自然杀伤细胞(natural killer cells,NK)表面表达杀伤免疫球蛋白样受体(killer immunologublin receptor,KIR,包括CD158a,CD158b,CD158e)及CD57分子的NK细胞亚群功能的异同,探讨具有不同功能状态的NK细胞亚群.方法:收集于北京大学血液病研究所行造血干细胞移植的健康供者10例,基于CD107a表达及细胞内因子干扰素-γ(interferon-γ,IFN-γ)的分泌,采用6色流式细胞学检测技术,检测不同NK细胞亚群NK细胞毒性及胞内细胞因子IFN-γ分泌功能的异同.结果:(1)表达CD57分子的NK细胞亚群占NK细胞的比例明显高于表达KIR分子的NK细胞亚群[(60.71%±5.71%)vs.(24.47%±3.95%),P〈0.001)],进一步用KIR分子及CD57分子将NK细胞亚群区分为KIR+CD57-,KIR+CD57+,KIR-CD57+,KIR-CD57-四群NK细胞亚群,KIR-CD57+ (43.03%±5.70%)及KIR-CD57- (32.45%±5.50%)所占比例相近(P=0.189),占NK比例最高,其次为KIR+CD57+(17.67%±3.39%)及KIR+CD57-(6.69%±0.95%);(2)进一步功能试验提示:表达CD57分子的NK细胞亚群与表达KIR分子的NK细胞亚群的杀伤活性及细胞因子IFN-γ分泌能力相近,二者差异无统计学意义,但明显低于CD56bri NK细胞(P=0.046,P=0.035),但与CD56dimNK细胞亚群杀伤活性及IFN-γ分泌能力相近;无论从NK亚群杀伤活性还是从NK亚群IFN-γ分泌活性排列,KIR-CD57-细胞最高(46.22%±9.24%和23.41%±5.82%),其杀伤活性与细胞因子分泌功能与CD56briNK细胞亚群相近(49.60%±9.26%和30.67%±8.0%),其次依次为KIR+CD57-细胞,KIR-CD57+细胞和KIR+CD57+细胞,后二者功能上差异无统计学意义.结论:CD57分子及KIR受体均与NK细胞功能密切相关,提示KIR-CD57-为早期NK细胞,CD57阳性细胞可能为终末分化NK细胞,而KIR+CD57-细胞为中间型NK细胞.
Objective:To explore the functional different natural killer (NK) cell subsets through the expression of killer immunoglobulin receptor (KIR) and CD57 on NK cells. Methods: From May 2012 to June 2012, the peripheral blood samples of 10 related healthy donors for hematopoietic stem cells transplantation were collected to analyze KIR, CD57 expression and the intracellular cytokines of interferon-γ( IFN-γ), and the CD107a secreted by NK cells through 6-colour flow cytometer to compare the cytokine secretion and cytotoxic function among different NK subset. Results: The expression of CD57 on NK cells were significantly higher than those of KIR on NK cells[ (60.71% ± 5.71% ) vs. (24.47% ±3.95% ), P 〈0.001 ]. All the NK cells were separated into KIR + CD57 -, KIR + CD57 +, KIR - CD57 + , KIR - CD57 - cells based on the expressions of KIR and CD57. The proportions of KIR-CD57 + NK cells (43.03% ±5.70%) and KIR-CD57-NK cells (32.45%±5.50%) among NK cells were comparable (P = 0. 189 ), and were higher than those of KIR + CD57 + NK cells (17.67% ±3.39% ) and KIR + CD57 - NK cells (6.69% ±0.95% ). Further functional experiments demonstrated that the cytotoxic function and IFN-γ cytokine secretion of CD57 + NK cells and KIR + NK cells were comparable, which were significantly lower than those of CD56b'i NK cells ( P = 0. 046 and 0.035, respectively), but were equal to those of CD56bira NK cells. The cytotoxic function and the IFN-γ secretion of KIR - CD57 - NK cells (46.22% ± 9.24% and 23.41% ±5.82% ) were significantly higher than those of the other NK subsets including KIR + CD57 - NK cells, KIR - CD57 + NK cells and KIR + CD57 + NK ceils, which were similar to those of CD56bri NK cells. The cytotoxic function and IFN-γ secretion of KIR + CD57 - NK cells were lower than those of KIR - CD57 - NK cells, but were higher than those of CD57 + NK cells, whether KIR positive or negative. The cytotoxic function and IFN-γ secreti