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1.
Infect Immun ; 68(5): 2566-72, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10768945

RESUMO

Complexes containing lipopolysaccharide (LPS) and three outer membrane proteins (OMPs) are released by gram-negative bacteria incubated in human serum and into the circulation in an experimental model of sepsis. The same OMPs are bound by immunoglobulin G (IgG) in the cross-protective antiserum raised to Escherichia coli J5 (anti-J5 IgG). This study was performed to identify the three OMPs. The 35-kDa OMP was identified as outer membrane protein A (OmpA) by immunoblotting studies using OmpA-deficient bacteria and recombinant OmpA protein. The 18-kDa OMP was identified as peptidoglycan-associated lipoprotein (PAL) based on peptide sequences from the purified protein and immunoblotting studies using PAL-deficient bacteria. The 5- to 9-kDa OMP was identified as murein lipoprotein (MLP) based on immunoblotting studies using MLP-deficient bacteria. The studies identify the OMPs released into human serum and into the circulation in an experimental model of sepsis as OmpA, PAL, and MLP.


Assuntos
Proteínas da Membrana Bacteriana Externa/análise , Escherichia coli/química , Lipoproteínas/análise , Peptidoglicano/análise , Proteoglicanas , Animais , Proteínas da Membrana Bacteriana Externa/imunologia , Escherichia coli/metabolismo , Proteínas de Escherichia coli , Humanos , Lipoproteínas/imunologia , Peptidoglicano/imunologia , Coelhos
2.
J Infect Dis ; 181(3): 1034-43, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10720528

RESUMO

Prior studies indicate that 3 bacterial outer-membrane proteins (OMPs) are released into serum associated with lipopolysaccharide (LPS) and are bound by IgG in antiserum to Escherichia coli J5 (anti-J5 IgG). The present studies analyzed the interaction of the OMPs with anti-J5 IgG and evaluated their release in an infected burn model of gram-negative sepsis. Affinity purification studies were performed on filtrates of bacteria incubated in human serum and plasma from rats with sepsis by use of O chain-specific anti-LPS IgG and anti-J5 IgG. All 3 OMPs were captured from septic rat blood by anti-LPS IgG. Release of OMPs into serum was highest for immature bacterial cultures and was increased by antibiotics in vitro and in vivo. Anti-J5 IgG selectively captured an 18-kDa OMP released into serum and into plasma from septic rats. The results raise the possibility that anti-J5 IgG may, in part, protect via anti-OMP antibodies.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Escherichia coli/imunologia , Infecções por Bactérias Gram-Negativas/metabolismo , Soros Imunes/metabolismo , Imunoglobulina G/metabolismo , Sepse/metabolismo , Animais , Antibacterianos/farmacologia , Anticorpos Monoclonais/imunologia , Proteínas da Membrana Bacteriana Externa/sangue , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Humanos , Camundongos , Coelhos , Ratos
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