摘要
目的利用虾原肌球蛋白及其Ig E单克隆抗体建立I型过敏反应体内外模型。方法采用等电点沉淀法制备刀额新对虾原肌球蛋白,利用杂交瘤技术制备抗虾原肌球蛋白Ig E单克隆抗体。建立虾原肌球蛋白攻击Ig E单抗致敏的RBL-2H3细胞模型,测定β-氨基己糖苷酶释放量。利用虾原肌球蛋白及其Ig E单抗建立被动全身过敏反应小鼠模型,监测抗原攻击后30 min内肛温的变化。结果 Ig E单抗致敏的RBL-2H3细胞在虾原肌球蛋白攻击后发生脱颗粒,β-氨基己糖苷酶释放量明显增加。虾原肌球蛋白及其Ig E单抗诱导了小鼠被动全身过敏反应,抗原攻击后小鼠肛温平均下降约1.44℃。结论虾原肌球蛋白及其Ig E单克隆抗体成功建立了I型过敏反应体内外模型。
Aim To develop the in vitro and in vivo models induced by shrimp tropomyosin( ST) and mono-clonal tropomyosin-specific murine IgE antibody ( anti-ST-IgE mAb) . Methods ST was purified from Metap-enaeusensis by an isoelectric precipitation method. The anti-ST-IgE mAb was obtained from hybridomas. After RBL-2 H3 cells were sensitized with anti-ST-IgE mAb and challenged with ST,β-hexosaminidase release was determined. Passive systemic anaphylaxis ( PSA ) was induced in mice and the rectal temperature was recor-ded after ST challenge within 30 min by a thermal probe. Results A significant increase ofβ-hexosamin-idase was observed in sensitized cells after ST chal-lenge. The average temperature drop after ST challenge was 1. 44℃ in PSA mice within 30 min. Conclusion The in vitro and in vivo models induced by ST and anti-ST-IgE mAb are established as an improvement of pres-ent models of type Ⅰ allergy.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第10期1422-1427,共6页
Chinese Pharmacological Bulletin
基金
国家科技部“重大新药创制”科技重大专项(No 2014ZX09201022-006)
北京市自然科学基金项目(No 7142112)