Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 Sep;60(9):3620–3629. doi: 10.1128/iai.60.9.3620-3629.1992

Molecular analysis of the serotyping antigens of Neisseria meningitidis.

I M Feavers 1, J Suker 1, A J McKenna 1, A B Heath 1, M C Maiden 1
PMCID: PMC257369  PMID: 1500170

Abstract

Molecular approaches to the rapid analysis of the serotyping antigens of Neisseria meningitidis, the class 2 and 3 outer membrane proteins (OMPs), were developed, evaluated, and used to study 12 antigenic variants of these proteins. A primer set for the polymerase chain reaction (PCR) amplification of the genes encoding these antigens was devised. Low-stringency amplification of meningococcal chromosomal DNA with this primer set resulted in the amplification of two products from each strain, whereas at higher stringencies only one product was amplified in most strains. Southern hybridization techniques and restriction analyses were used to differentiate the PCR products amplified at high stringencies from strains expressing class 2 or class 3 OMPs; these PCR products were further characterized by the determination of their nucleotide sequences, confirming that they represented the amplified class 2 and class 3 OMP genes. Analyses of these and other nucleotide sequences enabled the construction of a phenogram illustrating the interrelationships between Neisseria OMP genes. The comparative analysis of deduced amino acid sequences revealed conserved and variable regions of the proteins; the latter probably correspond to surface loops on the protein and hence are potentially exposed to the immune system. Further analyses of the primary structures of these related porins from Neisseria species enabled construction of models of the secondary structure of these antigens and comparison of these models with those previously published. The methods reported in the present work are rapid reproducible procedures for the analysis of antigenic variants of these proteins.

Full text

PDF
3624

Images in this article

3622Image
on p.3622
3623Image
on p.3623

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barlow A. K., Heckels J. E., Clarke I. N. The class 1 outer membrane protein of Neisseria meningitidis: gene sequence and structural and immunological similarities to gonococcal porins. Mol Microbiol. 1989 Feb;3(2):131–139. doi: 10.1111/j.1365-2958.1989.tb01802.x. [DOI] [PubMed] [Google Scholar]
  2. Butcher S., Sarvas M., Runeberg-Nyman K. Class-3 porin protein of Neisseria meningitidis: cloning and structure of the gene. Gene. 1991 Aug 30;105(1):125–128. doi: 10.1016/0378-1119(91)90523-e. [DOI] [PubMed] [Google Scholar]
  3. Carbonetti N. H., Simnad V. I., Seifert H. S., So M., Sparling P. F. Genetics of protein I of Neisseria gonorrhoeae: construction of hybrid porins. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6841–6845. doi: 10.1073/pnas.85.18.6841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carbonetti N. H., Sparling P. F. Molecular cloning and characterization of the structural gene for protein I, the major outer membrane protein of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9084–9088. doi: 10.1073/pnas.84.24.9084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cartwright K. A., Stuart J. M., Noah N. D. An outbreak of meningococcal disease in Gloucestershire. Lancet. 1986 Sep 6;2(8506):558–561. doi: 10.1016/s0140-6736(86)90124-8. [DOI] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Embley T. M. The linear PCR reaction: a simple and robust method for sequencing amplified rRNA genes. Lett Appl Microbiol. 1991 Sep;13(3):171–174. doi: 10.1111/j.1472-765x.1991.tb00600.x. [DOI] [PubMed] [Google Scholar]
  8. Feavers I. M., Heath A. B., Bygraves J. A., Maiden M. C. Role of horizontal genetic exchange in the antigenic variation of the class 1 outer membrane protein of Neisseria meningitidis. Mol Microbiol. 1992 Feb;6(4):489–495. doi: 10.1111/j.1365-2958.1992.tb01493.x. [DOI] [PubMed] [Google Scholar]
  9. Frasch C. E., Chapman S. S. Classification of Neisseria meningitidis group B into distinct serotypes. I. Serological typing by a microbactericidal method. Infect Immun. 1972 Jan;5(1):98–102. doi: 10.1128/iai.5.1.98-102.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frasch C. E. Vaccines for prevention of meningococcal disease. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S134–S138. doi: 10.1128/cmr.2.suppl.s134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frasch C. E., Zollinger W. D., Poolman J. T. Serotype antigens of Neisseria meningitidis and a proposed scheme for designation of serotypes. Rev Infect Dis. 1985 Jul-Aug;7(4):504–510. doi: 10.1093/clinids/7.4.504. [DOI] [PubMed] [Google Scholar]
  12. Gotschlich E. C., Seiff M. E., Blake M. S., Koomey M. Porin protein of Neisseria gonorrhoeae: cloning and gene structure. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8135–8139. doi: 10.1073/pnas.84.22.8135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hitchcock P. J. Unified nomenclature for pathogenic Neisseria species. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S64–S65. doi: 10.1128/cmr.2.suppl.s64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Klugman K. P., Gotschlich E. C., Blake M. S. Sequence of the structural gene (rmpM) for the class 4 outer membrane protein of Neisseria meningitidis, homology of the protein to gonococcal protein III and Escherichia coli OmpA, and construction of meningococcal strains that lack class 4 protein. Infect Immun. 1989 Jul;57(7):2066–2071. doi: 10.1128/iai.57.7.2066-2071.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maiden M. C., Suker J., McKenna A. J., Bygraves J. A., Feavers I. M. Comparison of the class 1 outer membrane proteins of eight serological reference strains of Neisseria meningitidis. Mol Microbiol. 1991 Mar;5(3):727–736. doi: 10.1111/j.1365-2958.1991.tb00743.x. [DOI] [PubMed] [Google Scholar]
  16. Markwell M. A., Haas S. M., Tolbert N. E., Bieber L. L. Protein determination in membrane and lipoprotein samples: manual and automated procedures. Methods Enzymol. 1981;72:296–303. doi: 10.1016/s0076-6879(81)72018-4. [DOI] [PubMed] [Google Scholar]
  17. McGuinness B., Barlow A. K., Clarke I. N., Farley J. E., Anilionis A., Poolman J. T., Heckels J. E. Deduced amino acid sequences of class 1 protein (PorA) from three strains of Neisseria meningitidis. Synthetic peptides define the epitopes responsible for serosubtype specificity. J Exp Med. 1990 Jun 1;171(6):1871–1882. doi: 10.1084/jem.171.6.1871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mocca L. F., Frasch C. E. Sodium dodecyl sulfate-polyacrylamide gel typing system for characterization of Neisseria meningitidis isolates. J Clin Microbiol. 1982 Aug;16(2):240–244. doi: 10.1128/jcm.16.2.240-244.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murakami K., Gotschlich E. C., Seiff M. E. Cloning and characterization of the structural gene for the class 2 protein of Neisseria meningitidis. Infect Immun. 1989 Aug;57(8):2318–2323. doi: 10.1128/iai.57.8.2318-2323.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Olyhoek T., Crowe B. A., Achtman M. Clonal population structure of Neisseria meningitidis serogroup A isolated from epidemics and pandemics between 1915 and 1983. Rev Infect Dis. 1987 Jul-Aug;9(4):665–692. doi: 10.1093/clinids/9.4.665. [DOI] [PubMed] [Google Scholar]
  21. Peltola H. Meningococcal disease: still with us. Rev Infect Dis. 1983 Jan-Feb;5(1):71–91. doi: 10.1093/clinids/5.1.71. [DOI] [PubMed] [Google Scholar]
  22. Poolman J. T., Hopman C. T., Zanen H. C. Immunogenicity of meningococcal antigens as detected in patient sera. Infect Immun. 1983 Apr;40(1):398–406. doi: 10.1128/iai.40.1.398-406.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Poolman J. T., Timmermans H. A., Teerlink T., Seid R. C., Jr Purification, cyanogen bromide cleavage, and amino terminus sequencing of class 1 and class 3 outer membrane proteins of meningococci. Infect Immun. 1989 Mar;57(3):1005–1007. doi: 10.1128/iai.57.3.1005-1007.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reingold A. L., Broome C. V., Hightower A. W., Ajello G. W., Bolan G. A., Adamsbaum C., Jones E. E., Phillips C., Tiendrebeogo H., Yada A. Age-specific differences in duration of clinical protection after vaccination with meningococcal polysaccharide A vaccine. Lancet. 1985 Jul 20;2(8447):114–118. doi: 10.1016/s0140-6736(85)90224-7. [DOI] [PubMed] [Google Scholar]
  25. Saukkonen K., Abdillahi H., Poolman J. T., Leinonen M. Protective efficacy of monoclonal antibodies to class 1 and class 3 outer membrane proteins of Neisseria meningitidis B:15:P1.16 in infant rat infection model: new prospects for vaccine development. Microb Pathog. 1987 Oct;3(4):261–267. doi: 10.1016/0882-4010(87)90059-3. [DOI] [PubMed] [Google Scholar]
  26. Schwartz B., Moore P. S., Broome C. V. Global epidemiology of meningococcal disease. Clin Microbiol Rev. 1989 Apr;2 (Suppl):S118–S124. doi: 10.1128/cmr.2.suppl.s118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Smith J. M., Dowson C. G., Spratt B. G. Localized sex in bacteria. Nature. 1991 Jan 3;349(6304):29–31. doi: 10.1038/349029a0. [DOI] [PubMed] [Google Scholar]
  28. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tinsley C. R., Heckels J. E. Variation in the expression of pili and outer membrane protein by Neisseria meningitidis during the course of meningococcal infection. J Gen Microbiol. 1986 Sep;132(9):2483–2490. doi: 10.1099/00221287-132-9-2483. [DOI] [PubMed] [Google Scholar]
  30. Tsai C. M., Frasch C. E., Mocca L. F. Five structural classes of major outer membrane proteins in Neisseria meningitidis. J Bacteriol. 1981 Apr;146(1):69–78. doi: 10.1128/jb.146.1.69-78.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Witholt B., Boekhout M., Brock M., Kingma J., Heerikhuizen H. V., Leij L. D. An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976 Jul;74(1):160–170. doi: 10.1016/0003-2697(76)90320-1. [DOI] [PubMed] [Google Scholar]
  32. Wolff K., Stern A. The class 3 outer membrane protein (PorB) of Neisseria meningitidis: gene sequence and homology to the gonococcal porin PIA. FEMS Microbiol Lett. 1991 Oct 1;67(2):179–185. doi: 10.1016/0378-1097(91)90351-a. [DOI] [PubMed] [Google Scholar]
  33. van der Ley P., Heckels J. E., Virji M., Hoogerhout P., Poolman J. T. Topology of outer membrane porins in pathogenic Neisseria spp. Infect Immun. 1991 Sep;59(9):2963–2971. doi: 10.1128/iai.59.9.2963-2971.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES