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Functional expression of the capsule polymerase of Neisseria meningitidis serogroup X: a new perspective for vaccine development.
- Source :
-
Glycobiology [Glycobiology] 2014 Feb; Vol. 24 (2), pp. 150-8. Date of Electronic Publication: 2013 Nov 20. - Publication Year :
- 2014
-
Abstract
- Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis and sepsis. A key feature in pathogenicity is the capsular polysaccharide (CPS) that prevents complement activation and thus supports bacterial survival in the host. Twelve serogroups characterized by immunologically and structurally different CPSs have been identified. Meningococcal CPSs elicit bactericidal antibodies and consequently are used for the development of vaccines. Vaccination against the epidemiologically most relevant serogroups was initially carried out with purified CPS and later followed by conjugate vaccines which consist of CPS covalently linked to a carrier protein. Of increasing importance in the African meningitis belt is NmX for which no vaccine is currently available. Here, we describe the molecular cloning, recombinant expression and purification of the capsule polymerase (CP) of NmX called CsxA. The protein expressed with N- and/or C-terminal epitope tags was soluble and could be purified to near homogeneity. With short oligosaccharide primers derived from the NmX capsular polysaccharide (CPSX), recombinant CsxA produced long polymer chains in vitro that in immunoblots were detected with NmX-specific antibodies. Moreover, the chemical identity of in vitro produced NmX polysaccharides was confirmed by NMR. Besides the demonstration that the previously identified gene csxA encodes the NmX CP CsxA, the data presented in this study pave the way for the use of the recombinant CP as a safe and economic way to generate the CPSX in vaccine developmental programs.
- Subjects :
- Antigens, Bacterial genetics
Antigens, Bacterial immunology
Bacterial Capsules enzymology
Cloning, Molecular
DNA-Directed DNA Polymerase immunology
DNA-Directed DNA Polymerase metabolism
Drug Discovery
Escherichia coli genetics
Escherichia coli metabolism
Gene Expression Regulation, Bacterial
Gene Expression Regulation, Enzymologic
Humans
Meningitis, Meningococcal immunology
Meningitis, Meningococcal prevention & control
Neisseria meningitidis enzymology
Recombinant Proteins genetics
Recombinant Proteins metabolism
Bacterial Capsules genetics
DNA-Directed DNA Polymerase genetics
Meningococcal Vaccines biosynthesis
Neisseria meningitidis genetics
Neisseria meningitidis immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1460-2423
- Volume :
- 24
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Glycobiology
- Publication Type :
- Academic Journal
- Accession number :
- 24259400
- Full Text :
- https://doi.org/10.1093/glycob/cwt102