title: Long-term anti-endotoxin/E. coli efficacy in mice transfected with AAV2/1-muBPI25 -muFcγ1.
abstract: Bactericidal/permeability increasing (BPI) is an antibiotic protein which kills Gram-negative bacteria and neutralizes endotoxin. We have previously developed a recombinant adeno-associated virus which contains human BPI amino acid residues 1-199 and Fc fragment of human IgG1 gene (AAV-hBPI-Fc) and shown that the recombinant virus can protect mice from lethal endotoxemia. However, whether AAV-hBPI-Fc can be used in vivo for the long term remains unclear. To address this, we established an adeno-associated virus-containing mouse BPI and Fc fragment genes (muBPI-Fc) and compared antigenicity of these recombinant proteins in murine models. Immunohistochemistry showed the expression of both fusion proteins at injected sites. ELISA and Western blotting showed that the muBPI-Fc protein was detected in serum up to 8 weeks after injection, without generation of autoantibodies against muBPI-Fc. In contrast, expressed hBPI-Fc protein was only detected on the 2nd week, whereas the autoantibody against hBPI-Fc protein occurred in serum from the 4th week to the end of study. muBPI-Fc also reduced production of proinflammatory cytokines and protected mice from endotoxemia and bacteremia. Our data showed that AAV-muBPI-Fc has potential long-term efficacy as an anti-endotoxin and has anti-bacterial activity in mice, suggesting the potential clinical application of AAV-hBPI-Fc, such as in endotoxin shock.
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Zhe
authors: Z
authors: Lv
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Yiqiang
authors: Y
authors: Fan
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Qing
authors: Q
authors: Xu
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Qingli
authors: Q
authors: Kong
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Chen
authors: C
authors: Li
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Zhenlong
authors: Z
authors: Liu
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China. wy_robin@ccmu.edu.cn.
authors: Wei
authors: W
authors: Wang
authors: The Department of Immunology and The Research Centre of Microbiome, Capital Medical University, Beijing, China.
authors: Yunqing
authors: Y
authors: An
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism; D001691:Biological Therapy / Q000379:methods*; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D004195:Disease Models, Animal; D004337:Drug Carriers; D019446:Endotoxemia / Q000517:prevention & control* / Q000628:therapy*; D004797:Enzyme-Linked Immunosorbent Assay; D004927:Escherichia coli Infections / Q000517:prevention & control* / Q000628:therapy*; D005260:Female; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D044967:Serum / Q000737:chemistry; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D004337:Drug Carriers; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C021623:bactericidal permeability increasing protein
keywords: Bactericidal/permeability increasing; anti-infection; bacteremia; endotoxemia; gram-negative bacteria