摘要
目的:制备一种以sLAG-3为佐剂的过敏原疫苗,并对该蛋白进行生物学活性测定,为建立一种高效安全的过敏原特异性免疫治疗奠定基础。方法:将sLAG-3和Fc段基因片段克隆入pcDNA3.1真核表达载体,电穿孔转染细胞,Western blotting检测生物活性。结果:成功构建可高度表达sLAG-3-Ig的真核表达载体,该蛋白可以明显抑制B细胞合成IgE的能力。结论:制备了一种以sLAG-3为佐剂的过敏原疫苗,该蛋白具有淋巴细胞抑制活性,从而可能建立一种高效安全的过敏原特异性免疫治疗手段。
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 Fe 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 DH5α, 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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第5期893-895,共3页
Chinese Journal of Pathophysiology
基金
陕西省自然科学基金资助项目(No.2002C2-16)
