The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and...The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and selection by methotrexate,the cell lines expressing the fusion protein were obtained.The recombinant protein was purified using cation\|exchange chromatography and its bioactivity was proved with bactericidal assays.展开更多
To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 (AAV2)-mediated BPI-Fcγ1 gene on protecti...To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 (AAV2)-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus (1 to 199 amino acidic residues) of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 (Fcγ1). Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular & Molecular Immunology. 2006;3(3):221-225.展开更多
文摘The fusion gene of BPI 23 and human Fcγ1 was obtained by PCR method,and the expression plasmid was constructed to express recombinant BPI 23 \|Fcγ1 fusion protein in CHO cells.After transfection with the plasmid and selection by methotrexate,the cell lines expressing the fusion protein were obtained.The recombinant protein was purified using cation\|exchange chromatography and its bioactivity was proved with bactericidal assays.
文摘To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 (AAV2)-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus (1 to 199 amino acidic residues) of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 (Fcγ1). Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular & Molecular Immunology. 2006;3(3):221-225.