Construction Of Inducible Costimulator Fusion Protein And Its Effect On Inflammation In A Murine Model Of Allergic Asthma

In this research,we cloned the extracellular region of human ICOS by RT-PCR from RNA of human peripheral blood mononuclearcytes purified cultured with PMA and ionomycin and Constructed a fusion proteins of extracellular portion of ICOS and heavy chain of mouse immunoglobulin with the eukaryotic expression technique. We also screened the biological activity of ICOS-mlg in vitro and in vivo.Then we evaluated the effect of ICOS-mlg on the inflammation in a murine allergic asthma.A recombinant human ICOS mutation with high receptor affinity activity was designed by Insight II software with the aid of a three-dimensional model and subcloned by QuikChangeSite朌 irected Mutagenesis Kit.We constructed a mutation fusion protein of the mutated ICOS and mouse lg and evaluated its biological activity with receptor affinity screening and mixture lymphocytes reaction. At last, to get to satisfy the need of production on large scale, we investigated a optimal condition of expression and fermentation of ICOS-lg engineering CHO cells.