食物致敏性评价RBL-2H3细胞模型的研究

A RBL-2H3 cell model for assessing the allergenicity of food proteins

目的研究大鼠嗜碱性白血病粒细胞(ra basophil leukemia cells,RBL-2H3细胞)用于食物致敏性评价的可行性。方法实验组和对照组动物血清按照不同倍数稀释,包括1:4、1:8、1:16、1:32、1:64、1:128、1:256和1:512,将稀释后的血清分别与RBL-2H3细胞孵育4 h,然后用MEM完全培养基重新培养24 h。最后,向细胞分别加入含有10μg/ml和100μg/ml OVA的MEM溶液,孵育45 min,测定上清液中β-氨基己糖苷酶含量和细胞内总的β-氨基己糖苷酶含量,计算β-氨基己糖苷酶释放率。结果对比不同的蛋白刺激浓度发现:100μg/ml OVA刺激后,稀释比例为1:4、1:8、1:16和1:32时,实验组和对照组差异有统计学意义(P<0.05,P<0.01);10μg/ml OVA刺激后,稀释比例仅1:4、1:8和1:16时,实验组和对照组差异有统计学意义(P<0.05,P<0.01)。对比不同的血清稀释倍数发现,1:4倍稀释时实验组β-氨基己糖苷酶释放率显著高于稀释比例1:256实验组,差异有统计学意义(P<0.05)。结论初步建立了RBL-2H3细胞致敏模型,以期用于食物致敏性评价中。

Objective To study the possibility of using the RBL-2H3 cell as a cell model for assessing the potential allergenicity of food proteins. Methods Sera of mice in the experimental group and control group were diluted as follows： 1：4, 1：8, 1：16, 1：32, 1：64, 1：128, 1：256 and 1：512. The RBL-2H3 cells were incubated with diluted sera for 4 hours, and then the cells were re-incubated with MEM complete medium for 24 hours. Lastly, the cells were respectively placed in the incubators with 10 μg/ml and 100 μg/ml ovalbumin （OVA） for 45 minutes. The beta-hexosaminidase in the supernatant and total intracellular beta-hexosaminidase were measured, and the release rate of beta-hexosaminidase was calculated. Results Comparison of different protein stimulation concentrations revealed that after stimulation with 100 μg/mL OVA, a statistically significant difference in the release rate of beta-hexosaminidase was observed between the experimental group and the control group when the serum diluton ratios were 1：4, 1：8, 1：16 and 1：32 respectively（P〈0.05 or P〈0.01）. After stimulation with 10μg/mL OVA, a statistically significant difference was found between the experimental group and the control group when the serum diluton ratios were 1：4, 1：8 and 1：16 respectively （P〈0.05 or P〈0.01）. Comparison of different serum dilution ratios in the experimental group indicated that the release rate of beta-hexosaminidase when the dilution ratio was 1：4 was higher than that when the dilution ratio was 1：256, showing a statistically significant difference （P〈0.05）. Conclusions A RBL-2H3 cell model for assessing the allergenicity of food proteins is initially established. It might be a valuable model for food allergy.