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. 1992 Aug;30(8):2047–2053. doi: 10.1128/jcm.30.8.2047-2053.1992

Production and characterization of monoclonal antibodies to type 8 lipooligosaccharide of Neisseria meningitidis.

X X Gu 1, C M Tsai 1, A B Karpas 1
PMCID: PMC265440  PMID: 1380009

Abstract

Eight monoclonal antibodies (MAbs) to lipooligosaccharides (LOSs) of Neisseria meningitidis were produced by immunizing mice with purified LOS from group A meningococcal strain A1. The specificities of the MAbs were examined by enzyme-linked immunosorbent assay (ELISA), immunodot assay, and ELISA inhibition by using the homologous A1 LOS, 12 immunotype LOSs of N. meningitidis (L1 through L12), and LOSs or lipopolysaccharides from other gram-negative bacteria. Two of the MAbs, 4385G7 (immunoglobulin G2b [IgG2b]) and 4387A5 (IgG2a), had the strongest reactivities with the homologous A1 LOS, moderate reactivities with the M978 (L8) LOS, but no reactivity with other LOSs. The other six MAbs (4 IgM and 2 IgG3) reacted with the A1 LOS and with several or many of the 12 LOSs. ELISA inhibition at 50% showed that the inhibitory activities of the LOSs from strains A1 and BB431 (a group B strain) to the specific MAb 4387A5 were about 10 to 20 times greater than that of the M978 (L8) LOS. When compared with MAb 2-1-L8 (L8) by Western blot (immunoblot) analysis and ELISA inhibition, the two specific MAbs recognized a different epitope in the 3.6-kDa LOSs of strains A1 and BB431. We propose that the new epitope is L8a, since the MAbs also reacted with the M978 (L8) LOS. The expression of the L8a epitope in the A1 LOS requires a few monosaccharide residues in its oligosaccharide moiety, and the fatty acid residues in its lipid A moiety also play a role. In a whole-cell ELISA, the two specific MAbs bound specifically to the homologous strain A1 and the L8 prototype strain M978 but not to any other LOS prototype strains. These results suggest that the two specific MAbs can be used for LOS typing of N. meningitidis.

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Selected References

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  1. Achtman M., Kusecek B., Morelli G., Eickmann K., Wang J. F., Crowe B., Wall R. A., Hassan-King M., Moore P. S., Zollinger W. A comparison of the variable antigens expressed by clone IV-1 and subgroup III of Neisseria meningitidis serogroup A. J Infect Dis. 1992 Jan;165(1):53–68. doi: 10.1093/infdis/165.1.53. [DOI] [PubMed] [Google Scholar]
  2. Apicella M. A., Bennett K. M., Hermerath C. A., Roberts D. E. Monoclonal antibody analysis of lipopolysaccharide from Neisseria gonorrhoeae and Neisseria meningitidis. Infect Immun. 1981 Dec;34(3):751–756. doi: 10.1128/iai.34.3.751-756.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Apicella M. A., Shero M., Jarvis G. A., Griffiss J. M., Mandrell R. E., Schneider H. Phenotypic variation in epitope expression of the Neisseria gonorrhoeae lipooligosaccharide. Infect Immun. 1987 Aug;55(8):1755–1761. doi: 10.1128/iai.55.8.1755-1761.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crothers D. M., Metzger H. The influence of polyvalency on the binding properties of antibodies. Immunochemistry. 1972 Mar;9(3):341–357. doi: 10.1016/0019-2791(72)90097-3. [DOI] [PubMed] [Google Scholar]
  5. Crowe B. A., Wall R. A., Kusecek B., Neumann B., Olyhoek T., Abdillahi H., Hassan-King M., Greenwood B. M., Poolman J. T., Achtman M. Clonal and variable properties of Neisseria meningitidis isolated from cases and carriers during and after an epidemic in The Gambia, West Africa. J Infect Dis. 1989 Apr;159(4):686–700. doi: 10.1093/infdis/159.4.686. [DOI] [PubMed] [Google Scholar]
  6. Dudas K. C., Apicella M. A. Selection and immunochemical analysis of lipooligosaccharide mutants of Neisseria gonorrhoeae. Infect Immun. 1988 Feb;56(2):499–504. doi: 10.1128/iai.56.2.499-504.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Estabrook M. M., Mandrell R. E., Apicella M. A., Griffiss J. M. Measurement of the human immune response to meningococcal lipooligosaccharide antigens by using serum to inhibit monoclonal antibody binding to purified lipooligosaccharide. Infect Immun. 1990 Jul;58(7):2204–2213. doi: 10.1128/iai.58.7.2204-2213.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Gotschlich E. C., Liu T. Y., Artenstein M. S. Human immunity to the meningococcus. 3. Preparation and immunochemical properties of the group A, group B, and group C meningococcal polysaccharides. J Exp Med. 1969 Jun 1;129(6):1349–1365. doi: 10.1084/jem.129.6.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Griffiss J. M., Schneider H., Mandrell R. E., Yamasaki R., Jarvis G. A., Kim J. J., Gibson B. W., Hamadeh R., Apicella M. A. Lipooligosaccharides: the principal glycolipids of the neisserial outer membrane. Rev Infect Dis. 1988 Jul-Aug;10 (Suppl 2):S287–S295. doi: 10.1093/cid/10.supplement_2.s287. [DOI] [PubMed] [Google Scholar]
  11. Gu X. X., Tsai C. M. Purification of rough-type lipopolysaccharides of Neisseria meningitidis from cells and outer membrane vesicles in spent media. Anal Biochem. 1991 Aug 1;196(2):311–318. doi: 10.1016/0003-2697(91)90472-6. [DOI] [PubMed] [Google Scholar]
  12. Hardy M. R., Townsend R. R., Lee Y. C. Monosaccharide analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection. Anal Biochem. 1988 Apr;170(1):54–62. doi: 10.1016/0003-2697(88)90089-9. [DOI] [PubMed] [Google Scholar]
  13. Ito J. I., Jr, Wunderlich A. C., Lyons J., Davis C. E., Guiney D. G., Braude A. I. Role of magnesium in the enzyme-linked immunosorbent assay for lipopolysaccharides of rough Escherichia coli strain J5 and Neisseria gonorrhoeae. J Infect Dis. 1980 Oct;142(4):532–537. doi: 10.1093/infdis/142.4.532. [DOI] [PubMed] [Google Scholar]
  14. Jennings H. J., Beurret M., Gamian A., Michon F. Structure and immunochemistry of meningococcal lipopolysaccharides. Antonie Van Leeuwenhoek. 1987;53(6):519–522. doi: 10.1007/BF00415511. [DOI] [PubMed] [Google Scholar]
  15. Kim J. J., Mandrell R. E., Griffiss J. M. Neisseria lactamica and Neisseria meningitidis share lipooligosaccharide epitopes but lack common capsular and class 1, 2, and 3 protein epitopes. Infect Immun. 1989 Feb;57(2):602–608. doi: 10.1128/iai.57.2.602-608.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kim J. J., Mandrell R. E., Hu Z., Westerink M. A., Poolman J. T., Griffiss J. M. Electromorphic characterization and description of conserved epitopes of the lipooligosaccharides of group A Neisseria meningitidis. Infect Immun. 1988 Oct;56(10):2631–2638. doi: 10.1128/iai.56.10.2631-2638.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Mandrell R. E., Griffiss J. M., Macher B. A. Lipooligosaccharides (LOS) of Neisseria gonorrhoeae and Neisseria meningitidis have components that are immunochemically similar to precursors of human blood group antigens. Carbohydrate sequence specificity of the mouse monoclonal antibodies that recognize crossreacting antigens on LOS and human erythrocytes. J Exp Med. 1988 Jul 1;168(1):107–126. doi: 10.1084/jem.168.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mandrell R. E., Lesse A. J., Sugai J. V., Shero M., Griffiss J. M., Cole J. A., Parsons N. J., Smith H., Morse S. A., Apicella M. A. In vitro and in vivo modification of Neisseria gonorrhoeae lipooligosaccharide epitope structure by sialylation. J Exp Med. 1990 May 1;171(5):1649–1664. doi: 10.1084/jem.171.5.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mandrell R. E., Zollinger W. D. Lipopolysaccharide serotyping of Neisseria meningitidis by hemagglutination inhibition. Infect Immun. 1977 May;16(2):471–475. doi: 10.1128/iai.16.2.471-475.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Phillips N. J., John C. M., Reinders L. G., Gibson B. W., Apicella M. A., Griffiss J. M. Structural models for the cell surface lipooligosaccharides of Neisseria gonorrhoeae and Haemophilus influenzae. Biomed Environ Mass Spectrom. 1990 Nov;19(11):731–745. doi: 10.1002/bms.1200191112. [DOI] [PubMed] [Google Scholar]
  23. Poolman J. T., Hopman C. T., Zanen H. C. Immunochemical characterization of Neisseria meningitidis serotype antigens by immunodiffusion and SDS-polyacrylamide gel electrophoresis immunoperoxidase techniques and the distribution of serotypes among cases and carriers. J Gen Microbiol. 1980 Feb;116(2):465–473. doi: 10.1099/00221287-116-2-465. [DOI] [PubMed] [Google Scholar]
  24. Rouse D. A., Morris S. L., Karpas A. B., Probst P. G., Chaparas S. D. Production, characterization, and species specificity of monoclonal antibodies to Mycobacterium avium complex protein antigens. Infect Immun. 1990 May;58(5):1445–1449. doi: 10.1128/iai.58.5.1445-1449.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Salih M. A., Danielsson D., Bäckman A., Caugant D. A., Achtman M., Olcén P. Characterization of epidemic and nonepidemic Neisseria meningitidis serogroup A strains from Sudan and Sweden. J Clin Microbiol. 1990 Aug;28(8):1711–1719. doi: 10.1128/jcm.28.8.1711-1719.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schneider H., Griffiss J. M., Boslego J. W., Hitchcock P. J., Zahos K. M., Apicella M. A. Expression of paragloboside-like lipooligosaccharides may be a necessary component of gonococcal pathogenesis in men. J Exp Med. 1991 Dec 1;174(6):1601–1605. doi: 10.1084/jem.174.6.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schneider H., Griffiss J. M., Mandrell R. E., Jarvis G. A. Elaboration of a 3.6-kilodalton lipooligosaccharide, antibody against which is absent from human sera, is associated with serum resistance of Neisseria gonorrhoeae. Infect Immun. 1985 Dec;50(3):672–677. doi: 10.1128/iai.50.3.672-677.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schneider H., Hale T. L., Zollinger W. D., Seid R. C., Jr, Hammack C. A., Griffiss J. M. Heterogeneity of molecular size and antigenic expression within lipooligosaccharides of individual strains of Neisseria gonorrhoeae and Neisseria meningitidis. Infect Immun. 1984 Sep;45(3):544–549. doi: 10.1128/iai.45.3.544-549.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sugasawara R. J., Prato C., Sippel J. E. Monoclonal antibodies against Neisseria meningitidis lipopolysaccharide. Infect Immun. 1983 Dec;42(3):863–868. doi: 10.1128/iai.42.3.863-868.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsai C. M., Boykins R., Frasch C. E. Heterogeneity and variation among Neisseria meningitidis lipopolysaccharides. J Bacteriol. 1983 Aug;155(2):498–504. doi: 10.1128/jb.155.2.498-504.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tsai C. M., Civin C. I. Eight lipooligosaccharides of Neisseria meningitidis react with a monoclonal antibody which binds lacto-N-neotetraose (Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc). Infect Immun. 1991 Oct;59(10):3604–3609. doi: 10.1128/iai.59.10.3604-3609.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  34. Tsai C. M., Mocca L. F., Frasch C. E. Immunotype epitopes of Neisseria meningitidis lipooligosaccharide types 1 through 8. Infect Immun. 1987 Jul;55(7):1652–1656. doi: 10.1128/iai.55.7.1652-1656.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamasaki R., Schneider H., Griffiss J. M., Mandrell R. Epitope expression of gonococcal lipooligosaccharide (LOS). Importance of the lipoidal moiety for expression of an epitope that exists in the oligosaccharide moiety of LOS. Mol Immunol. 1988 Aug;25(8):799–809. doi: 10.1016/0161-5890(88)90116-2. [DOI] [PubMed] [Google Scholar]
  36. Zollinger W. D., Mandrell R. E. Importance of complement source in bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide. Infect Immun. 1983 Apr;40(1):257–264. doi: 10.1128/iai.40.1.257-264.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zollinger W. D., Mandrell R. E. Outer-membrane protein and lipopolysaccharide serotyping of Neisseria meningitidis by inhibition of a solid-phase radioimmunoassay. Infect Immun. 1977 Nov;18(2):424–433. doi: 10.1128/iai.18.2.424-433.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zollinger W. D., Mandrell R. E. Type-specific antigens of group A Neisseria meningitidis: lipopolysaccharide and heat-modifiable outer membrane proteins. Infect Immun. 1980 May;28(2):451–458. doi: 10.1128/iai.28.2.451-458.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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