RESUMEN
AIM: Production of water soluble protein-containing antigens from various strains of S. pneumoniae during cultivation in complete and semi-synthetic culture media as well as selection of strains with cross antigenic activity. MATERIALS AND METHODS: S. pneumoniae 3, 6A, 6B, 14, 10A, 18A, 19A, 19F, 23F serotype strains were cultivated in brain-heart broth and semi-synthetic medium with addition of aminopeptide for 24 hours at 37 degrees C for the production of water soluble antigens. The antigens were obtained by a method of triple water extraction from acetone dried microbial cells. Chemical composition of preparations, electrophoresis mobility of protein-containing components of preparations and cross antigenic activity in gel immune diffusion reaction by using rabbit hyperimmune sera were studied. RESULTS: In studies of 10 pneumococcus strains from various serotypes a method of microbial cell inactivation by acetone was selected that allows to produce preparations with high protein content (25.5 - 53.1%). Electrophoretic separation of the preparations revealed difference in the preparations obtained from various pneumococcus strains in the layout of major protein lines in the 8 - 95 kDa range. The most virulent and immunogenic S. pneumoniae strain that during cultivation in semi-synthetic medium was characterized by intraspecies cross antigenic activity and in gel immune diffusion reacted with all the studied sera against 3, 14, 18C, 23F serotype strains was selected. CONCLUSION: The study resulted in the selection of a technologically simple method of production of pneumococcus antigens with high protein content and showed that only 1 of the studied preparations produced from a virulent strain with poorly expressed S. pneumoniae capsule during cultivation in semi-synthetic medium has the highest cross antigenic activity.
Asunto(s)
Antígenos Bacterianos , Proteínas Bacterianas , Streptococcus pneumoniae , Animales , Antígenos Bacterianos/química , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/aislamiento & purificación , Proteínas Bacterianas/química , Proteínas Bacterianas/inmunología , Proteínas Bacterianas/aislamiento & purificación , Medios de Cultivo/química , Conejos , Solubilidad , Streptococcus pneumoniae/química , Streptococcus pneumoniae/crecimiento & desarrollo , Streptococcus pneumoniae/inmunología , Agua/químicaRESUMEN
The study of cross-reactions between healthy pulmonary tissue antigens and Moraxella catarrhalis with the use of SDS-electrophoresis and immunoblotting revealed that in the component of healthy pulmonary tissue with a mol. wt. of 40 kD epitopes existed to which antibodies were produced, capable of cross reaction with the components of M. catarrhalis with a mol. wt. of 35 kD and 70 kD. In addition, the presence of cross-reactions between cytokeratin-8, protein contained in healthy pulmonary tissue, and M. catarrhalis antigens was established.
Asunto(s)
Antígenos/inmunología , Pulmón/inmunología , Moraxella catarrhalis/inmunología , Antígenos Bacterianos/inmunología , Reacciones Cruzadas , Epítopos/química , Epítopos/inmunología , Humanos , Immunoblotting , Queratinas/inmunología , Peso Molecular , Moraxella catarrhalis/químicaRESUMEN
Data on specific antibodies to the bacterial antigens of Yersinia enterocolitica (serovars 03, 09) and Proteus mirabilis in rheumatoid arthritis (RA) patients are presented. According to the data of some researchers, these organisms play some role in the pathogenesis of RA. The highest level of IgG antibodies to Y. enterocolitica antigens of both serovars was noted. The activity of specific IgG antibodies to serovar 03 was higher than that to serovar 09. In the sera of RA patients with P. mirabilis antgens the activity of IgM antibodies was higher.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Artritis Reumatoide/sangre , Artritis Reumatoide/etiología , Proteus mirabilis/inmunología , Yersinia enterocolitica/inmunología , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Infecciones por Proteus/complicaciones , Serotipificación , Yersiniosis/complicaciones , Yersinia enterocolitica/clasificaciónRESUMEN
Cytotoxicity of hybrid plasmide pGalI was associated with the genome site measuring for 1.1.mD and flanked by the recognizing sites of Eco RI restrictase. The cytotoxic effect of pGalI might be limited not only by plasmide pMB9 but also by other plasmides constructed on the base of pMBI replicone. Limitation of the cytotoxicity occurred due to a decrease in amount of the pGalI copies. Capacity of the plasmides studied to limit the pGalI cytotoxicity enabled to clone them as cryptic plasmides under conditions of both simultaneous with pGalI transformation of E. coli and the subsequent selection of clones with the Gal+ phenotype.