RESUMO
Bacterial adherence is a complex phenomenon involving specific interactions between receptors, including matricial fibronectin, and bacterial ligands. We show here that fibronectin and outer membrane proteins of Pseudomonas fluorescens were able to inhibit adherence of P. fluorescens to fibronectin-coated wells. We identified at least six fibronectin-binding proteins with molecular masses of 70, 55, 44, 37, 32 and 28 kDa. The presence of native (32 kDa) and heat-modified forms (37 kDa) of OprF was revealed by immuno-analysis and the 44-kDa band was composed of three proteins, their N-terminal sequences showing homologies with Pseudomonas aeruginosa porins (OprD, OprE1 and OprE3).
Assuntos
Proteínas da Membrana Bacteriana Externa , Proteínas de Bactérias , Fibronectinas/metabolismo , Porinas/metabolismo , Pseudomonas fluorescens/metabolismo , Aderência Bacteriana , Western Blotting , Porinas/genética , Homologia de Sequência de AminoácidosAssuntos
Iridoviridae , Lepidópteros/microbiologia , Animais , Antígenos Virais/análise , Linhagem Celular , Efeito Citopatogênico Viral , DNA Viral/análise , Hidrolases/metabolismo , Insetos/microbiologia , Iridoviridae/análise , Iridoviridae/enzimologia , Iridoviridae/fisiologia , Iridoviridae/ultraestrutura , Lipídeos/análise , Proteínas Virais/análise , Replicação ViralRESUMO
Pseudomonas fluorescens is a highly heterogeneous species and includes both avirulent strains and clinical strains involved in nosocomial infections. We previously demonstrated that clinical strain MFN1032 has hemolytic activity involving phospholipase C (PlcC) and biosurfactants (BSs), similar to that of the opportunistic pathogen Pseudomonas aeruginosa. When incubated under specific conditions, MFN1032 forms translucent phenotypic variant colonies defective in hemolysis, but not necessarily in PlcC. We analyzed eight variants of the original strain MFN1032 and found that they clustered into two groups. Mutations of genes encoding the two-component regulatory system GacS/GacA are responsible for phenotypic variation in the first group of variants. These group 1 variants did not produce secondary metabolites and had impaired biofilm formation. The second group was composed of hyperflagellated cells with enhanced biofilm capacity: they did not produce BSs and were thus unable to swarm. Artificial reduction of the intracellular level of c-di-GMP restored the ability to form biofilm to levels shown by the wild type, but production of BSs was still repressed. Phenotypic variation might increase the virulence potential of this strain.
Assuntos
Mutação , Infecções por Pseudomonas/microbiologia , Pseudomonas fluorescens/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes , Humanos , Fenótipo , Pseudomonas fluorescens/classificação , Pseudomonas fluorescens/genética , Tensoativos/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
There is some debate about the potential survival of Pseudomonas fluorescens at temperatures above 37 degrees C and its consequences for infectious potential, owing to the heterogeneity of clinical strains. Seven clinical strains growing at 37 degrees C or more were submitted for polyphasic identification; 2 were identified as Pseudomonas mosselii and 4 were precisely characterized as P. fluorescens bv. I or II. The binding indexes on glial cells of the strains identified as P. fluorescens bv. I and P. mosselii were compared with that of a reference psychrotrophic strain, P. fluorescens MF37 (bv. V). Clinical P. fluorescens had a similar adherence potential range than strain MF37. Conversely, the binding indexes for P. mosselii strains were 3 times greater than that for strain MF37. These data, and those obtained by comparing the cytotoxic activities of P. fluorescens clinical strains, suggest the existence of different virulence mechanisms, leading either to a low infectious form or to a microorganism with cytotoxic activity in the same range as that of P. mosselii or even Pseudomonas aeruginosa.
Assuntos
Infecções por Pseudomonas/microbiologia , Pseudomonas fluorescens/classificação , Pseudomonas fluorescens/fisiologia , Apoptose , Aderência Bacteriana , Técnicas de Cultura de Células , DNA Bacteriano/genética , DNA Ribossômico/genética , Humanos , L-Lactato Desidrogenase/metabolismo , Neuroglia/microbiologia , Nitritos/metabolismo , Pseudomonas fluorescens/genética , Pseudomonas fluorescens/isolamento & purificação , RNA Ribossômico 16S/genética , Sorotipagem , TemperaturaRESUMO
Monoclonal antibodies (McAbs) obtained against Iridovirus type 6 (CIV) were characterized by Western blotting and/or immunoprecipitation. Seven McAbs were found to be strongly reactive with viral polypeptides of molecular weights 16 K and 18 K by Western blotting. Two McAbs were directed against a complex composed of 30 K, 50 K, and 100 K polypeptides, but failed to react with either of these free polypeptides. This finding could explain the faint reactivity of these McAbs in Western blotting and immunoprecipitation. The reactivity of the other McAbs with their antigenic determinants is also discussed.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Iridoviridae/imunologia , Proteínas Virais/imunologia , Anticorpos Antivirais/imunologia , Precipitação Química , Epitopos , Técnicas de Imunoadsorção , Substâncias Macromoleculares , Relação Estrutura-AtividadeRESUMO
The surface antigens of Iridovirus type 6 (CIV) were characterized by crossed immunoelectrophoresis. Using different solubilization techniques, up to seven antigenic determinants corresponding to four viral structural polypeptides were identified.
Assuntos
Antígenos de Superfície/análise , Antígenos Virais/análise , Iridoviridae/imunologia , Proteínas Virais/imunologia , Epitopos , Imunoeletroforese Bidimensional , Iridoviridae/isolamento & purificação , Iridoviridae/ultraestrutura , Proteínas Virais/análise , Proteínas Estruturais ViraisRESUMO
The behaviour of different Invertebrate cell lines iridovirus type 6 (CIV) infection was compared. The results allow us to distinguish at least four types of cellular systems: non-permissive systems (a. albopictus), semi-permissive systems (L. dispar) and two types of permissive systems in which the viral replication cycle is complete A. aegypti (slow replication cycle) and C. fumiferana (rapid replication cycle)