IgG1类抗IgE Fc单克隆抗体抑制人肥大细胞脱颗粒作用研究  

Inhibition of Anti-IgE Fc monoclonal antibodies of IgG1 class on the degranulation of mast cells

作　　者：周翔鱼[1] 曹羿堃 李娜[1] 储晨晨 华冰[1] 王佳[1] 张秋丽[2] ZHOU Xiangyu;CAO Yikun;LI Na;CHU Chenchen;HUA Bing;WANG Jia;ZHANG Qiuli(Gladstone Institute of Neurological Disease,The First People’s Hospital of Shizuishan,Shizuishan 753600,China;Department of Pediatric,The First People′s Hospital of Shizuishan,Shizuishan 753600,China)

机构地区：[1]宁夏石嘴山市第一人民医院神经疾病研究所,宁夏石嘴山753200 [2]宁夏石嘴山市第一人民医院儿科,宁夏石嘴山753200

出　　处：《宁夏医学杂志》2022年第4期305-307,F0003,共4页Ningxia Medical Journal

基　　金：宁夏自然科学基金资助项目(NZ17250)。

摘　　要：目的探究IgG1类抗IgE Fc单克隆抗体对人肥大细胞脱颗粒作用的影响。方法采用骨髓杂交瘤技术制备IgG1类大鼠抗人IgE Fc单克隆抗体,ELISA法鉴定杂交瘤细胞分泌的特异性抗体亚类;进一步体外培养人LAD2肥大细胞,分别以Compound 48/80、Compound 48/80+hum-IgE-Fc、Compound 48/80+anti-IgE-Fc mAb、Compound 48/80+IgE-Fc+anti-IgE-Fc mAb致敏LAD2肥大细胞48 h,Compound 48/80刺激45 min后甲苯胺蓝染色,收各组上清液,用ELISA检测HIS和TPS的释放浓度,Western blot检测各组TPS和CD32b分子表达水平。结果成功制备2株分泌IgG1类Anti-IgE-Fc mAb的杂交瘤细胞株,与对照组相比,Compound 48/80促进LAD2细胞脱颗粒效应,HIS和TPS的释放浓度明显增加(P<0.05),且肥大细胞TPS蛋白表达上调。Compound 48/80+hum-IgE-Fc及Compound 48/80+anti-IgE-Fc mAb组较Compound48/80组LAD2细胞颗粒释放浓度HIS和TPS及CD32b蛋白差异无统计学意义(P>0.05);Compound48/80联合hum-IgE-Fc和anti-IgE-Fc mAb时显著抑制肥大细胞脱颗粒反应,同时肥大细胞TPS蛋白表达下调,表面抑制性分子CD32b表达上调(P<0.05)。结论IgG1类大鼠抗人IgE Fc单克隆抗体在游离IgE-Fc存在时可抑制人肥大细胞脱颗粒,这种作用可能主要由CD32b介导。Objective To study the effects of the Anti-IgE Fc monoclonal antibodies of IgG1 class on the degranulation of human mast cells.Methods Anti-IgE Fc monoclonal antibodies were prepared by hybridoma technique,and the specific antibody subclasses secreted by hybridoma cells was identified by ELISA.Further,human mast cells LAD2 were cultured in vitro.LAD2 were sensitized with Compound48/80,Compound48/80+hum-IgE-Fc,Compound48/80+anti-IgE-Fc mAb,Compound48/80+IgE-Fc+anti-IgE-Fc mAb for 48h and after compound 48/80 were sensitized for 45min,the release concentration of His and TPS and toluidine blue staining of LAD2 cells were detected by ELISA in supernatant of each group.The protein expression of TPS and CD32b in each group was detected by Western blot.Rusults Two strains of Anti-IgE Fc monoclonal antibodies of IgG1 class were successfully prepared.Compared with the control group,compound 48/80 significantly promoted the degranulation effect of LAD2 mast cells,the release concentrations of HIS and TPS in supernatant were significantly increased(P<0.05),and the expression of TPS protein in mast cells was upregulated,but CD32b had no significant change.Compared with compound48/80 group,there was no significant difference in the release concentrations of HIS and TPS and the expression of CD32b protein(P>0.05)in compound 48/80+hum-IgE FC and compound 48/80+anti IgE FC mAb groups.Compound 48/80 combined with hum-IgE-Fc and anti-IgE Fc mAb significantly inhibited the degranulation of mast cells,while the expression of TPS protein in mast cells was down-regulated and the expression of surface inhibitory molecule CD32b was up-regulated(P<0.05).Conclusion Anti-IgE Fc monoclonal antibodies of IgG1 class may inhibit the degranulation of human mast cells in the presence of free IgE-Fc,and the effect may be mainly mediated by CD32b.

关 键 词：人肥大细胞脱颗粒 IgE Fc抗原 抗IgE Fc单克隆抗体 CD32b 

分 类 号：R392.8[医药卫生—免疫学]