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J Med Microbiol 58 (2009), 196-208; DOI: 10.1099/jmm.0.004820-0
© 2009 Society for General Microbiology
ISSN 0022-2615

Neisseria meningitidis antigen NMB0088: sequence variability, protein topology and vaccine potential

Gretel Sardiñas1,{dagger}, Daniel Yero1,2,{dagger}, Yanet Climent1,2, Evelin Caballero1, Karem Cobas1 and Olivia Niebla1

1 Meningococcal Research Department, Division of Vaccines, Center for Genetic Engineering and Biotechnology, Avenue 31, Cubanacan, Habana 10600, Cuba

2 Department of Molecular Biology, Division of Biotechnology, Finlay Institute, Avenue 27, La Lisa, Habana 11600, Cuba

Correspondence
Gretel Sardiñas
gretel.sardinas{at}cigb.edu.cu
Daniel Yero
dyero{at}finlay.edu.cu

Received July 7, 2008
Accepted October 4, 2008

The significance of Neisseria meningitidis serogroup B membrane proteins as vaccine candidates is continually growing. Here, we studied different aspects of antigen NMB0088, a protein that is abundant in outer-membrane vesicle preparations and is thought to be a surface protein. The gene encoding protein NMB0088 was sequenced in a panel of 34 different meningococcal strains with clinical and epidemiological relevance. After this analysis, four variants of NMB0088 were identified; the variability was confined to three specific segments, designated VR1, VR2 and VR3. Secondary structure predictions, refined with alignment analysis and homology modelling using FadL of Escherichia coli, revealed that almost all the variable regions were located in extracellular loop domains. In addition, the NMB0088 antigen was expressed in E. coli and a procedure for obtaining purified recombinant NMB0088 is described. The humoral immune response elicited in BALB/c mice was measured by ELISA and Western blotting, while the functional activity of these antibodies was determined in a serum bactericidal assay and an animal protection model. After immunization in mice, the recombinant protein was capable of inducing a protective response when it was administered inserted into liposomes. According to our results, the recombinant NMB0088 protein may represent a novel antigen for a vaccine against meningococcal disease. However, results from the variability study should be considered for designing a cross-protective formulation in future studies.

Abbreviations: CREE, Correia repeat-enclosed element; DRVs, dried–reconstituted vesicles; i.n., intranasal; i.p., intraperitoneal; OMVs, outer-membrane vesicles; TM, transmembrane; VCN, vancomycin/colistin/nystatin; VRs, variable regions.

{dagger}These authors contributed equally to this work.

The GenBank/EMBL/DDBJ accession numbers for the new nmb0088 gene sequences are EU645484–EU645512.






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