抗体修饰抑制性Ly49受体对小鼠NK细胞功能的影响

Effects on activity of mouse NK cells by inhibitory Ly49 receptors blockade

目的:观察单克隆抗体修饰小鼠NK细胞的抑制性Ly49受体对NK细胞功能的影响。方法:免疫磁性分选近交系小鼠C57BL/6J脾脏NK细胞,以小鼠肿瘤细胞YAC-1和M1细胞为研究对象,观察单克隆抗体封闭NK细胞的抑制性受体Ly49C/I后,对YAC-1和M1细胞的杀伤作用和对M1细胞克隆形成的影响,并观察抗体修饰对C57BL/6J小鼠脾脏NK细胞与BALB/C小鼠脾单个核细胞之间单向混合淋巴细胞反应的影响。结果:抗体修饰后,NK细胞对M1细胞的杀伤能力增强(10.7%±0.5%vs84.4%±2.9%),并与抗体浓度呈现量效关系(P<0.05);而对YAC-1细胞的杀伤作用则无明显的变化,仅0μg/ml组与60μg/ml组有统计学差异(P<0.05)。抗体作用后,对M1细胞克隆形成的抑制率增高(8.2%±2.2%vs94.0%±3.3%),并与抗体浓度呈现量效关系(P<0.05)。混合淋巴细胞反应中反应细胞的增殖指数降低(0.398±0.025vs0.128±0.014),并具有统计学差异(P<0.05)。结论:以抗体修饰抑制性Ly49受体有助于活化NK细胞,提高抗肿瘤能力。

Objective： To observe the effects on antitumor activity of mouse NK cells and the mixed lymphocyte reaction with blocking the interaction between the inhibitory Ly49 receptor on mouse NK cell and MHC-1 molecules on target cells with monoclonal antibody. Methods： NK cells were isolated from splenocytes of C57BL/6J mouse with immunomagnetic method. YAC-1 and M1 cells were used as target cells,the cytotoxicity of NK cells was examined with mixed lymphocyte reaction before and after treatment of anti-Ly49 monoclonal antibody. Results： The cytotoxicity of NK cells to M1 cell was significantly augmented （10. 7%±0. 5% vs 84. 4%± 2.9%）,and the clone-forming capacity of M1 cell was inhibited remarkably（8. 2%±2.2% vs 94.0% ±3.3 %）with the increase of concentration of the antibody（0～60μg/ml） （P〈0. 05）. The difference of cytotoxicity to YAC-1 cells was only found between 0 μg/ml and 60 μg/ml groups （P〈0. 05）. In mixed lymphocyte reaction, after pretreated with antibody, the proliferation of splenocytes of BALB/C mouse was inhibited （0. 398±0. 025 vs 0. 128± 0. 014）, and statistical difference was observed only between antibody groups of 0 μg/ml and 60 μg/ml （P〈0. 05）. Conclusion： Blocking the inhibitory Ly49 receptor on NK cells with monoclonal antibody augments the cytotoxicity to target cells.