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Towards the immunoproteome of Neisseria meningitidis.

Mendum TA, Newcombe J, McNeilly CL, McFadden J - PLoS ONE (2009)

Bottom Line: This is in part due to the difficulties in developing a, cross-protective vaccine that is effective against all serogroups, including serogroup B meningococci.Although a number of outer membrane antigens were identified the majority of the antigens were cytoplasmic, with roles in cellular processes and metabolism.When recombinant proteins were expressed and used to raise sera in mice, none of the antigens elicited a positive SBA result, however flow cytometry did demonstrate that some, including the ribosomal protein, RplY were localised to the neisserial cell surface.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom.

ABSTRACT
Despite the introduction of conjugated polysaccharide vaccines for many of the Neisseria meningitidis serogroups, neisserial infections continue to cause septicaemia and meningitis across the world. This is in part due to the difficulties in developing a, cross-protective vaccine that is effective against all serogroups, including serogroup B meningococci. Although convalescent N. meningitidis patients develop a natural long-lasting cross-protective immunity, the antigens that mediate this response remain unknown. To help define the target of this protective immunity we identified the proteins recognized by IgG in sera from meningococcal patients by a combination of 2D protein gels, western blots and mass spectrometry. Although a number of outer membrane antigens were identified the majority of the antigens were cytoplasmic, with roles in cellular processes and metabolism. When recombinant proteins were expressed and used to raise sera in mice, none of the antigens elicited a positive SBA result, however flow cytometry did demonstrate that some, including the ribosomal protein, RplY were localised to the neisserial cell surface.

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2D gels of total N. meningitidis proteins.Total N. meningitidis proteins separated by 2D gel electrophoresis using (a) a non-linear pI 4–7 1st dimension and (b) a non-linear pI 6–9 1st dimension. Gels were silver stained and replica gels western blotted with patient sera. Spots that were recognised by one or more sera on western blots are circled. Spots whose identity was determined are numbered in black, those that remain unidentified in grey.
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pone-0005940-g001: 2D gels of total N. meningitidis proteins.Total N. meningitidis proteins separated by 2D gel electrophoresis using (a) a non-linear pI 4–7 1st dimension and (b) a non-linear pI 6–9 1st dimension. Gels were silver stained and replica gels western blotted with patient sera. Spots that were recognised by one or more sera on western blots are circled. Spots whose identity was determined are numbered in black, those that remain unidentified in grey.

Mentions: Proteins extracted from N. meningitidis L91543 were separated by 2D gel electrophoresis and western blotted with sera from acute and convalescent patients (Table 1). Up to 473 separate protein spots could be distinguished on the 2D gels (Fig 1). Eighty eight of these 473 spots bound sufficient IgG from one or more of the patient sera to be detected on western blots (Fig 2).


Towards the immunoproteome of Neisseria meningitidis.

Mendum TA, Newcombe J, McNeilly CL, McFadden J - PLoS ONE (2009)

2D gels of total N. meningitidis proteins.Total N. meningitidis proteins separated by 2D gel electrophoresis using (a) a non-linear pI 4–7 1st dimension and (b) a non-linear pI 6–9 1st dimension. Gels were silver stained and replica gels western blotted with patient sera. Spots that were recognised by one or more sera on western blots are circled. Spots whose identity was determined are numbered in black, those that remain unidentified in grey.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2691954&req=5

pone-0005940-g001: 2D gels of total N. meningitidis proteins.Total N. meningitidis proteins separated by 2D gel electrophoresis using (a) a non-linear pI 4–7 1st dimension and (b) a non-linear pI 6–9 1st dimension. Gels were silver stained and replica gels western blotted with patient sera. Spots that were recognised by one or more sera on western blots are circled. Spots whose identity was determined are numbered in black, those that remain unidentified in grey.
Mentions: Proteins extracted from N. meningitidis L91543 were separated by 2D gel electrophoresis and western blotted with sera from acute and convalescent patients (Table 1). Up to 473 separate protein spots could be distinguished on the 2D gels (Fig 1). Eighty eight of these 473 spots bound sufficient IgG from one or more of the patient sera to be detected on western blots (Fig 2).

Bottom Line: This is in part due to the difficulties in developing a, cross-protective vaccine that is effective against all serogroups, including serogroup B meningococci.Although a number of outer membrane antigens were identified the majority of the antigens were cytoplasmic, with roles in cellular processes and metabolism.When recombinant proteins were expressed and used to raise sera in mice, none of the antigens elicited a positive SBA result, however flow cytometry did demonstrate that some, including the ribosomal protein, RplY were localised to the neisserial cell surface.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Health and Medical Sciences, University of Surrey, Guildford, United Kingdom.

ABSTRACT
Despite the introduction of conjugated polysaccharide vaccines for many of the Neisseria meningitidis serogroups, neisserial infections continue to cause septicaemia and meningitis across the world. This is in part due to the difficulties in developing a, cross-protective vaccine that is effective against all serogroups, including serogroup B meningococci. Although convalescent N. meningitidis patients develop a natural long-lasting cross-protective immunity, the antigens that mediate this response remain unknown. To help define the target of this protective immunity we identified the proteins recognized by IgG in sera from meningococcal patients by a combination of 2D protein gels, western blots and mass spectrometry. Although a number of outer membrane antigens were identified the majority of the antigens were cytoplasmic, with roles in cellular processes and metabolism. When recombinant proteins were expressed and used to raise sera in mice, none of the antigens elicited a positive SBA result, however flow cytometry did demonstrate that some, including the ribosomal protein, RplY were localised to the neisserial cell surface.

Show MeSH
Related in: MedlinePlus