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.
Keywords: beta and gamma chain(hFcεRIαβγ); food allergy assessment; human high affinity receptor containing alpha; lentivirus; rat basophil leukemia(RBL)-2H3.