RESUMO
Incubation of Yersinia pseudotuberculosis cells grown on a solid medium with pseudotuberculous diagnostic bacteriophage for 20 min at 37oC led to a significant decrease in the concentration of both components of the system. This effect was absent when the bacteria were grown in a fluid medium. At the same time, this incubation regimen promoted vesicle formation and typical morphological changes in bacteria grown in both surface and suspension cultures. Co-incubation of the bacteriophage with suspension of vesicles isolated from the suspension culture of Y. pseudotuberculosis grown at 10oC (but not 37oC) led to a decrease in plaque-forming activity of the bacteriophage.
Assuntos
Bacteriófagos/fisiologia , Vesículas Extracelulares/virologia , Yersinia pseudotuberculosis/virologia , Bacteriófagos/ultraestrutura , Meios de Cultura/química , Vesículas Extracelulares/ultraestrutura , Interações Microbianas , Microscopia Eletrônica , Yersinia pseudotuberculosis/química , Yersinia pseudotuberculosis/ultraestruturaRESUMO
The effect of treatment of Yersinia pseudotuberculosis cells with antibodies of various specificities on adhesiveness of pseudotuberculosis bacteriophage was analyzed by competitive inhibition technique. Bacteriophage adsorption to bacteria was sterically inhibited by monoclonal antibodies to protein epitopes of Y. pseudotuberculosis outer membrane. These results suggest that receptors of pseudotuberculosis diagnostic bacteriophage are localized on the LPS core of microbial cell.
Assuntos
Anticorpos Monoclonais/imunologia , Bacteriófagos/metabolismo , Lipopolissacarídeos/metabolismo , Receptores Virais/imunologia , Ligação Viral , Yersinia pseudotuberculosis/metabolismo , Infecções por Yersinia pseudotuberculosis/virologiaRESUMO
A hybridoma panel producing monoclonal antibodies to immunochemically non-identical antigenic epitopes of the protein nature located in outer membrane of Yersinia pseudotuberculosis was obtained. It was revealed that the previously identified B-antigen protecting laboratory animals from experimental plague was detected using both monoclonal antibodies against mentioned protein determinants and the determinants of lipopolysaccharide O-side chains. The B-antigen is a component of the Y. pseudotuberculosis outer membrane, egested in the form of the vesicles (OMVs) by bacteria cultivated in fluid nutrient medium.
Assuntos
Antígenos de Bactérias/química , Antígenos de Bactérias/imunologia , Epitopos/imunologia , Yersinia pseudotuberculosis/imunologia , Anticorpos Monoclonais/imunologia , Coloides , Epitopos/química , Ouro , Hibridomas/imunologia , Lipopolissacarídeos/imunologia , Microscopia Eletrônica de Transmissão , Peptídeo Hidrolases/química , Peptídeo Hidrolases/metabolismo , Temperatura , Yersinia pseudotuberculosis/crescimento & desenvolvimentoRESUMO
A study of the influence of exogenous factors on the immunochemical activity of the bacterium Yersinia pseudotuberculosis and lipopolysaccharide preparations isolated from bacteria was performed using monoclonal antibodies. It was shown that the hybridomas that were obtained in this work produce antibodies against different and, most likely, species-specific epitopes associated with lipopolysaccharide O side chains. The antibody concentrations produced increased with a decrease in the temperature, at which the bacteria were cultivated. An inhibitory effect of proteinase K, pepsin, and trypsin on the immunochemical activity of bacterial cells, determined using a solid-phase enzyme immunoassay, was demonstrated. Treatment with sodium periodate showed no uniform effect on the reactions between monoclonal antibodies and antigens (lipopolysaccharides and microbial cells), as adjudged by an immunoassay, which is most likely a consequence of the different localization of lipopolysaccharide epitopes recognized by the antibodies from four hybridomas.
Assuntos
Anticorpos Monoclonais/química , Antígenos de Bactérias/química , Antígenos de Superfície/química , Epitopos/química , Yersinia pseudotuberculosis/química , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/isolamento & purificação , Especificidade de Anticorpos , Antígenos de Bactérias/imunologia , Antígenos de Superfície/imunologia , Western Blotting , Reações Cruzadas , Endopeptidase K/química , Epitopos/imunologia , Hibridomas/imunologia , Técnicas Imunoenzimáticas , Lipopolissacarídeos/química , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Pepsina A/química , Ácido Periódico/química , Tripsina/química , Yersinia pseudotuberculosis/imunologiaRESUMO
The water-soluble polysaccharide fractions SAcI and SAclI were isolated from rowan tree stem calli. The SAcI fraction was shown to contain compounds belonging to the arabinogalactan II group. The SAcII fraction, termed sorban, was found to contain pectic polysaccharides that make up the protopectin complex of the cell wall callusand that had a high content of galacturonic acid residues together with neutral sugars characteristic of rhamnogalacturonan I (RG-I) polysaccharides. Using methylation assays, some structural features of the ramified region of sorban were elucidated. The backbone of sorban was found to consist of 1,4-α-D-galacto-pyranosyluronic acid residues. The neutral sugars are represented by 1,6-linked galactopyranose and 1,5-linked arabinofuranose residues as the primary constituents, as well as 1,6-linked mannopyranose and 1,4-linked glucopyranose and xylopyranose residues. Callus growth was shown to be accompanied by nearly constant quantities of galacturonic acid and neutral sugar residues in sorban (fraction SAcII) from the rowan stem callus.
