RESUMEN
AIM: To determine sialic acids and O-acetyl groups content in Yersinia pestis and Vibrio cholerae antigens in order to establish their association with biological activity. MATERIALS AND METHODS: The following antigens of Y. pestis EV NIIEG strain--capsular antigen (F1), major somatic antigen (MSA), lipopolysaccharide (LPS), Pla-protease, allergen pestin PP--as well as O-antigens (O-AG) of V. cholerae serogroups O1 and O139 were used in the study. Sialic acids were identified by the thiobarbituric method, and O-acetyl groups--according to Alicino. Specific polysaccharides in the MSA and O-antigens were detected by the immunodiffusion assay. RESULTS: Sialic acids were found in LPS, Pla-protease, allergen pestin PP, and all cholera O-AG; their absence was demonstrated in MSA and F1. O-acetyl groups were identified in cholera O-AG of both studied serogroups as well as in LPS, Pla-protease, MSA and pestin PP of Y. pestis. Tendency to correlation between O-acetyl groups content in MSA and serological activity titer was observed. CONCLUSION: Sialic acids and O-acetyl groups identified in carbohydrate-containing antigens of Y. pestis and V. cholerae could be characterized as reaction-active markers of pathogenetic mechanisms of cholera and plague infections as well as immunochemical activity of microbial polysaccharides.
Asunto(s)
Ácidos Siálicos/análisis , Vibrio cholerae , Yersinia pestis , Alérgenos/análisis , Alérgenos/aislamiento & purificación , Animales , Proteínas Bacterianas/análisis , Proteínas Bacterianas/aislamiento & purificación , Biomarcadores/análisis , Técnicas de Cultivo de Célula , Cólera/inmunología , Cólera/patología , Humanos , Inmunodifusión , Antígenos O/análisis , Antígenos O/aislamiento & purificación , Peste/inmunología , Peste/patología , Activadores Plasminogénicos/análisis , Activadores Plasminogénicos/aislamiento & purificación , Roedores , Pruebas Serológicas , Ácidos Siálicos/inmunología , Relación Estructura-Actividad , Vibrio cholerae/química , Vibrio cholerae/inmunología , Yersinia pestis/química , Yersinia pestis/inmunologíaRESUMEN
Some properties of the structure of Y. pestis capsular antigen macromolecules have been studied. The aminoacid composition of F1 protein, the aminoacid sequence of the N-terminal fragment of antigen polipeptide chain were determined. Some peculiarities in the dissociation of capsular antigen macromolecules have been studied. The formation of the product resulting from unterminated thermodissociation of F1 protein oligomeric form, consisting of four subunits, has been registered. The aspects of F1 protein association are discussed.
Asunto(s)
Antígenos Bacterianos/análisis , Proteínas Bacterianas/análisis , Yersinia pestis/análisis , Secuencia de Aminoácidos , Fenómenos Químicos , Química Física , Peso Molecular , Conformación Proteica , Yersinia pestis/inmunologíaRESUMEN
In this work the influence of Y. pestis antigens (capsular antigen, mouse toxin, lipopolysaccharide, pesticin), as well as the products of gene expression localized on three Y. pestis resident plasmids, have been studied. Capsular antigen and some antigens coded for by plasmid pCad have been shown to possess essential activity in the initiation of the synthesis of interleukin-1. Mouse toxin and the products of plasmid pPst produce the opposite effect.
Asunto(s)
Antígenos Bacterianos/inmunología , Inmunización , Interleucina-1/biosíntesis , Yersinia pestis/inmunología , Animales , Cápsulas Bacterianas/inmunología , Toxinas Bacterianas/inmunología , Bacteriocinas/inmunología , Células Cultivadas , Lipopolisacáridos/inmunología , Ratones , Ratones Endogámicos CBA , Plásmidos/inmunología , Bazo/inmunología , Timo/inmunologíaRESUMEN
The influence of Y. pestis phospholipase D on the physiological state of leukocytes in the blood of guinea pigs was studied in vivo by flow impulse fluorometry with the use of fluorochrome acridine orange. During the first hours of observation the intensity of leukocyte fluorescence increased due to a rise in the number of polymorphonuclear leukocytes and changes in the permeability of cell membranes. Further changes in the intensity of the fluorescence of the material under study after 24 hours of observation occurred due to the appearance of activated lymphocytes in the blood stream. The processes normalized by day 21. The reaction of blood leukocytes to phospholipase D was specific in comparison with the reaction to capsular antigen, "mouse" toxin, lipopolysaccharide and the main somatic antigen.
Asunto(s)
Antígenos Bacterianos/toxicidad , Leucocitos/efectos de los fármacos , Leucocitos/inmunología , Fosfolipasa D/toxicidad , Yersinia pestis/enzimología , Yersinia pestis/inmunología , Animales , Citometría de Flujo/instrumentación , Citometría de Flujo/métodos , Cobayas , Leucocitos/química , Fosfolipasa D/aislamiento & purificación , Peste/sangre , Peste/etiología , Peste/inmunología , Factores de Tiempo , Yersinia pestis/patogenicidadRESUMEN
Antiplague immunization of mice causes an increase in the number of T-suppressors in their thymus and spleen; this increase is especially pronounced after injection of a low-immunogenic dose of the vaccine strain. T-suppressors specific to Yersinia pestis antigens were detected in the thymus on day 3 after a single injection and on day 14 after two injections of the vaccine strain.
Asunto(s)
Peste/inmunología , Linfocitos T Reguladores/inmunología , Animales , Especificidad de Anticuerpos/inmunología , Antígenos Bacterianos/inmunología , Relación Dosis-Respuesta Inmunológica , Femenino , Tolerancia Inmunológica/inmunología , Inmunidad Celular/inmunología , Masculino , Ratones , Ratones Endogámicos , Vacuna contra la Peste/inmunología , Bazo/inmunología , Timo/inmunología , Yersinia pestis/inmunologíaRESUMEN
Immunization with live plague vaccine has been shown to give no protection to thymectomized mice from subcutaneous challenge with Y. pestis virulent strain. Under the action of the vaccine or individual Y. pestis antigens (fraction I) the functional and morphological activation of thymocytes and macrophages is observed, more pronounced in C57BL/6 mice and less pronounced in CBA mice. Y. pestis antigenic preparations (fractions I and II, pesticin) act as T-cell mitogens and are thus capable of inducing the in vitro proliferation of thymocytes. At the same time the in vivo action of fraction II induces a decrease in the level of lymphocytes in the peripheral blood of mice and the destruction of lymphocytes in their thymus and spleen.
Asunto(s)
Antígenos Bacterianos/inmunología , Peste/inmunología , Yersinia pestis/inmunología , Animales , Antígenos Bacterianos/toxicidad , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/toxicidad , Haplotipos/inmunología , Inmunidad Celular/inmunología , Inmunización , Ratones , Ratones Endogámicos , Fagocitos/inmunología , Linfocitos T/inmunología , Timo/inmunologíaRESUMEN
Two to 8 years after vaccination, specific antibodies to capsular antigen (F1) and lipopolysaccharide (LPS) are found in the blood sera of but 10 to 20% of subjects 1-4 times epicutaneously immunized with live antipest vaccine. Only regular continuous (for at least 7-15 years) antipest vaccination leads to the production of antibodies to the main antigens of Yersinia pestis, detectable in remote periods in 75.9 +/- 4.5% of vaccinees. It is noteworthy that the number of positive responses and titer of antibodies to LPS is appreciably higher than to F1. Detection of antibodies to Y. pestis is recommended for the retrospective diagnosis of pestis in both humans and animals in natural foci of this infection. Competitive enzyme immunoassay on nitrocellulose membranes with monoclonal antibodies is to be preferred for the purpose.
Asunto(s)
Anticuerpos Antibacterianos/análisis , Antígenos Bacterianos/inmunología , Cápsulas Bacterianas/inmunología , Lipopolisacáridos/inmunología , Vacuna contra la Peste/administración & dosificación , Yersinia pestis/inmunología , Animales , Humanos , Ratones , Ratones Endogámicos BALB CRESUMEN
Lymphocytic blast-cell transformation (LBCT) is shown to be related both to the genotype of a macroorganism and to the genetically determined expression of individual antigens by the plague microbe. The correlation of LBCT with the level of antiinfectious resistance suggests that this index is of high informative value and that it can be effectively used for testing the cellular immunity and the characteristically biological properties of the plague causative agent and its antigens.
Asunto(s)
Antígenos Bacterianos/inmunología , Activación de Linfocitos/inmunología , Yersinia pestis/inmunología , Yersinia pestis/patogenicidad , Animales , Relación Dosis-Respuesta Inmunológica , Genotipo , Inmunización , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Vacuna contra la Peste/inmunología , Bazo/inmunología , Factores de Tiempo , Virulencia , Yersinia pestis/genéticaAsunto(s)
Alérgenos/inmunología , Peste/diagnóstico , Yersinia pestis/inmunología , Fenómenos Químicos , Química , Humanos , Pronasa , Pruebas CutáneasAsunto(s)
5'-Nucleotidasa/inmunología , Antígenos Bacterianos/inmunología , Cápsulas Bacterianas/inmunología , Macrófagos Peritoneales/inmunología , Ratones Endogámicos C57BL/inmunología , Ratones Endogámicos CBA/inmunología , Yersinia pestis/inmunología , Animales , Cruzamientos Genéticos , Macrófagos Peritoneales/enzimología , Masculino , Ratones , Factores de TiempoAsunto(s)
Antígenos Bacterianos/inmunología , Epítopos/inmunología , Regulación Bacteriana de la Expresión Génica/inmunología , Activación de Linfocitos/inmunología , Yersinia pestis/inmunología , Animales , Inmunización , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Virulencia/inmunología , Yersinia pestis/patogenicidadRESUMEN
It is shown that immunomodulator of bacterial origin salmozan causes alternations of sensitivity to mouse toxin in mice CBA. A correlation exists between the sensitivity to mouse toxin and the level of 5-nucleotidase of peritoneal exudate macrophages.
Asunto(s)
5'-Nucleotidasa/análisis , Toxinas Bacterianas/inmunología , Macrófagos/enzimología , Yersinia pestis , Adyuvantes Inmunológicos/administración & dosificación , Animales , Endotoxinas/administración & dosificación , Masculino , Ratones , Ratones Endogámicos CBA , Modelos Estructurales , Polisacáridos Bacterianos/administración & dosificaciónRESUMEN
Immunochemical analysis of 2 polysaccharide-containing structures of the lypopolysaccharide of the plague causative agent (main somatic antigen and lipopolysaccharide) isolated from K-1 strain and a number of its antibiotic resistant mutants was carried out. It was shown that development of resistance to streptomycin alone or its combination with monomycin did not cause detectable changes in the monosaccharide composition and serological properties of the cultures tested. More significant changes associated with development of complex resistance, i.e. K-1 (Strr leads to Penr leads to Tetr) were accompanied by a decrease in the content of hexozamine and serological activity of the main somatic antigen determining the O-specificity of the lipopolysaccharide. Defective changes in the monosaccharide composition and serological properties of both the main somatic antigen and the polysaccharide were observed in the yellow variant of the streptomycin resistant mutant K-1.
Asunto(s)
Antibacterianos/farmacología , Antígenos Bacterianos/aislamiento & purificación , Mutación/efectos de los fármacos , Polisacáridos Bacterianos/aislamiento & purificación , Yersinia pestis/inmunología , Farmacorresistencia Microbiana , Monosacáridos/análisis , Polisacáridos Bacterianos/inmunología , Yersinia pestis/efectos de los fármacosRESUMEN
The investigation reveals different influence of the plague microbe's fraction 1 polysaccharide-protein complex and of it's purified protein on the 5'-nucleotidase activity and on the chemiluminescence response of peritoneal macrophages. Both of these metabolic indexes were found to be dependent on the dose of fraction 1 and duration of time till examination.