Preparation and evaluation of vibrio cholerae O1 EL Tor Ogawa lipopolysaccharide-tetanus toxoid conjugates.

The lipopolysaccharide (LPS) of Vibrio cholerae is considered one of the most important antigens from the point of view of immunogenicity in these bacteria. We have undertaken detoxification of this LPS by basic hydrolysis and the resultant amine groups were used for their conjugation to tetanus toxoid as carrier protein using carbodiimide-mediated coupling. The resulting conjugates were inoculated in Balb/c mice for immunogenicity studies. The anti-LPS IgG and vibriocidal antibodies were measured in serum. The antigenicity of this conjugated was evaluated by ELISA, with serums of humans vaccinated with a strain genetically modified. The conjugated elicited: high titers of IgG anti-LPS, high titers of vibriocidal antibodies and there was recognition of LPS by antibodies from cholerae immunised human serum. These results show that the conjugated LPS obtained by us, could be evaluated like a potential vaccine for human use.

New method for obtaining conjugated vaccines.

We describe a new method to obtain conjugates against Neisseria meningitidis serogroups A, B, C, Vibrio cholera, and Salmonella typhi and their immunogenicity in Balb/c mice. The saccharides were activated by basic hydrolysis with the generation of amine groups in the saccharidic chain, and these groups were linked to carboxyl groups of tetanus toxoid by via carbodiimida-mediated reaction. The resultant conjugates were administered to mice for the immunogenicity studies. The pirogenicity of LPS was measured by LAL assay. The anti-saccharide IgG, IgG1, and IgG2a antibodies were evaluated. A significant decrease in the pirogenicity of LPS after basic hydrolysis treatment was observed. The conjugates elicited higher titers of anti-polysaccharides or anti-LPS IgG, IgG1, and IgG2a in conjugates than in unconjugated saccharides. The results indicate that we have a new method for obtaining conjugated vaccines and we have demonstrated that after conjugation there was a change in the responses for all saccharides, from thymus-independent to thymus-dependent responses.

Validación de un Elisa para la cuantificación de IgG antipolisacárido capsular de neisseria meningitidis serogrupos C en sueros de ratones / Validation of the ELISA for cuantifying capsular anti-polysaccharide IgG of Neisseria meningitidis of the serogroup C in mice sera

Se desarrolló un ensayo inmunoenzimático de fase sólida (ELISA) indirecto para cuantificar anticuerpos IgG específicos antipolisacárido C en ratón, utilizando un prerrecubrimiento con Poli-L-lisina y luego el polisacárido capsular de Neisseria Meningitidis serogrupo C (Instituto Finlay, La Habana, Cuba), para evaluar la respuesta inmune contra este componente en candidatos vacunales en estudios preclínicos. Como conjugado se utilizó anti-IgG ratón conjugado a fosfatasa alcalina, el cual se une a los anticuerpos antipolisacárido C produccidos en ratones...(AU)

Fragmentación Polisacárido de Neisseria Meningitidis Serogrupo C para uso en Vacunas Conjugadas / Size fragmentation of Neisseria meningitidis grupo C polysaccharide for their use in conjugated vaccine

Las infecciones meningocócicas son una causa importante de morbilidad y mortalidad en el mundo. Los serogrupos B y C son responsables de la mayoría de los casos reportados en muchos países e incluso en países desarrollados. Las vacunas meningocócicas que contienen al polisacárido capsular purificado del meningococo C inducen en adultos protección por la presencia de anticuerpos bactericidas en sueros pero son pobremente inmunogénicos en niños pequeños y pueden inducir tolerancia. La inmunogenicidad de los polisacáridos o sus fragmentos puede ser mejorada por la conjugación a proteínas transportadoras...(AU)