RESUMEN
Two cyclic forms of the Enterobacterial Common Antigen were isolated from Escherichia coli O157:H(-). These antigenic determinants were purified from the biomass through extensive chemical, enzymatic and chromatographic procedures whereas MALDI MS spectrometry indicated their cyclic nature with a polymerization degree of 4 or 5. The two species, denoted as ECA(CYC-4) and ECA(CYC-5), were assigned by NMR and showed no further substitution with other appendages such as acetyl groups as usually described for similar cyclic antigens from other Enterobacteriaceae.
Asunto(s)
Antígenos Bacterianos/química , Antígenos Bacterianos/metabolismo , Escherichia coli O157/química , Polímeros/químicaRESUMEN
The broadly neutralizing antibody 2G12 binds a fairly conserved cluster of oligomannose sugars on the HIV surface glycoprotein gp120, which has led to the hypothesis that these sugars pose potential vaccine targets. Here, we present the chemical analysis, antigenicity, and immunogenicity of a bacterial lipooligosaccharide (LOS) comprised of a manno-oligosaccharide sequence analogous to the 2G12 epitope. Antigenic similarity of the LOS to oligomannose was evidenced by 2G12 binding to the LOS and the inability of sera elicited against synthetic oligomannosides, but incapable of binding natural oligomannose, to bind the LOS. Immunization with heat-killed bacteria yielded epitope-specific serum antibodies with the capacity to bind soluble gp120. Although these sera did not exhibit specific anti-HIV activity, our data suggest that this LOS may find utility as a template for the design of glycoconjugates to target HIV.
Asunto(s)
Vacunas contra el SIDA/metabolismo , Anticuerpos Neutralizantes/metabolismo , Infecciones por VIH/prevención & control , Lipopolisacáridos/metabolismo , Rhizobium/metabolismo , Vacunas contra el SIDA/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/metabolismo , Anticuerpos Neutralizantes/inmunología , Secuencia de Carbohidratos , Epítopos/inmunología , Proteína gp120 de Envoltorio del VIH/inmunología , Proteína gp120 de Envoltorio del VIH/metabolismo , Infecciones por VIH/inmunología , Humanos , Lipopolisacáridos/inmunología , Ratones , Datos de Secuencia Molecular , Unión ProteicaRESUMEN
The structures of the capsular polysaccharides (CPSs) of the two clinical isolates Acinetobacter baumannii SMAL and MG1 were elucidated. Hot phenol/water extractions of the dry biomasses, followed by enzymatic digestions and repeated ultracentrifugations led to the isolation of polysaccharides that were negative in Western blot analysis utilizing an anti-lipid A antibody, thus proving that they were not the LPS O-antigens but CPSs. Their structures were established on the basis of NMR spectroscopy and GC-MS analyses. The A. baumannii MG1 CPS consisted of a linear aminopolysaccharide with acyl substitution heterogeneity at the N-4 amino group of QuipN4N: 4)-α-d-GlcpNAc-(1â4)-α-l-GalpNAcA-(1â3)-ß-d-QuipNAc4NR-(1â R=3-hydroxybutyrryl or acetyl. The repeating unit of the CPS produced by strain SMAL is a pentasaccharide, already reported for the O-antigen moiety from A. baumannii strain ATCC 17961:
Asunto(s)
Acinetobacter baumannii/química , Polisacáridos Bacterianos/aislamiento & purificación , Infecciones por Acinetobacter/microbiología , Acinetobacter baumannii/inmunología , Secuencia de Carbohidratos , Cromatografía de Gases y Espectrometría de Masas , Datos de Secuencia Molecular , Antígenos O/química , Polisacáridos Bacterianos/químicaRESUMEN
Rhizobium rubi, strain DSM 30149, is a Gram negative phytopathogenic bacterium which produces a linear polysaccharide with the following repeating unit: This new structure was determined by spectroscopical and chemical methods. It presents similar lipophilic features reported for another strain of R. rubi. These contrast with features already known for capsular polysaccharide species from symbiontic members of the Rhizobiaceae family, namely highly anionic polymers.