Cloning,expression and characterization of sLAG-3-Ig / 中国病理生理杂志

ABSTRACT

AIM: To obtain a vaccine with sLAG-3 as immunoadjuvant and investigate its biologic activity in order to establish the safe and effective way for asthma as one of the specific immunotherapy.METHODS: The coding sequence of LAG-3 was amplified by polymerase chain reaction,the expression vector pcDNA-sLAG-3-Ig was constructed by inserting the PCR products of sLAG-3 and Fc sequence of IgG.With electroporation transfection,pcDNA-sLAG-3-Ig was transfected into COS-7 cells and its biologic activity was investigated by Western blotting analysis.RESULTS: By temperature induction,the LAG-3-Ig was highly expressed in E.coli DH5?.LAG-3-Ig fusion protein was observed by SDS-PAGE and Western blotting,the results showed that the LAG-3-Ig protein was an antagonist of the IL-4-induced synthesis of IgE in B cells.CONCLUSION: A new vaccine with sLAG-3 as immunoadjuvant was obtained.It could inhibit synthesis of IgE in B cells.Thus,LAG-3-Ig would be hopeful to establish the safe and effective way for asthma as one of the specific immunotherapy.