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Published on: November 1, 2014
[Construction and identification of stable hFcεRIαβγ/RBL-2H3 cells for food allergy assessment]
Chen Chen1, Qin Zhuo1, Yongning Li2
1Key Laboratory of Trace Element Nutrition of National Health Commission(NHC) , National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Objective:
To establish an rat basophil leukemia(RBL)-2H3 cell line stably expressing human high affinity receptor containing alpha, beta and gamma chain(hFcεRIαβγ), in order to provide experimental materials for evaluating allergenicity of food.
Methods:
The lentivirus was transfected into RBL-2H3 cells, and the mRNA expression of hFcεRIαβγ in cells was detected by real-time PCR and the protein expression of hFcεRIα was detected by flow cytometry.
Results:
Sequencing result showed that recombinant lentiviral vector GV367-hFcεRIαβγ was successfully constructed. According to the result of experiments, lentivirus could effectively infect RBL-2H3 cells. The mRNA of hFcεRIαβγ and protein levels of hFcεRIα in RBL-2H3 cells were successfully overexpressed.
Conclusion:
The hFcεRIαβγ/RBL-2H3 cells were preliminarily constructed, which could be binded with human IgE and further used in the evaluation system of food allergy, compared to RBL-2H3 cells.
