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. 1995 Sep;63(9):3683–3692. doi: 10.1128/iai.63.9.3683-3692.1995

The tetrasaccharide L-alpha-D-heptose1-->2-L-alpha-D-heptose1--> 3-L-alpha-D-heptose1-->(3-deoxy-D-manno-octulosonic acid) and phosphate in lipid A define the conserved epitope in Haemophilus lipopolysaccharides recognized by a monoclonal antibody.

S Borrelli 1, O Hegedus 1, D H Shaw 1, P E Jansson 1, A A Lindberg 1
PMCID: PMC173510  PMID: 7543887

Abstract

A murine monoclonal antibody, MAHI 3 (immunoglobulin G2b), that is broadly reactive with Haemophilus influenzae lipopolysaccharides (LPSs) but nonreactive with all enterobacterial LPSs tested was generated by fusing mouse myeloma cells with spleen cells of BALB/c mice immunized with azide-killed H. influenzae RM.7004. MAHI 3 bound to all H. influenzae, all other human Haemophilus spp., all Bordetella pertussis and Bordetella parapertussis, and all Aeromonas spp. tested but not to any Neisseria or Moraxella catarrhalis strains, as determined by enzyme immunoassay, colony dot immunoblotting, and immunoblotting. In an inhibition enzyme immunoassay, MAHI 3 reacted with all 45 H. influenzae LPSs tested but not with the LPS from the rough mutant I69 Rd-/b+, which has only 3-deoxy-D-manno-octulosonic acid (P) [Kdo(P)] and lipid A. The antibody was not inhibited by H. influenzae lipid A or lipid-free polysaccharide isolated after mild acid hydrolysis. Only native LPSs show positive inhibitory activity, indicating that part of lipid A is involved in the binding of MAHI 3. From the results, it is indicated that the structural element recognized by MAHI 3 is Hep alpha 1-->2Hep alpha 1-->3Hep alpha 1-->Kdo together with part of lipid A, including the phosphate.

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Selected References

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  1. Banoub J. H., Choy Y. M., Michon F., Shaw D. H. Structural investigations on the core oligosaccharide of Aeromonas hydrophila (Chemotype II) lipopolysaccharide. Carbohydr Res. 1983 Apr 1;114(2):267–276. doi: 10.1016/0008-6215(83)88193-2. [DOI] [PubMed] [Google Scholar]
  2. Brade H., Galanos C. Common lipopolysaccharide specificity: new type of antigen residing in the inner core region of S- and R-form lipopolysaccharides from different families of gram-negative bacteria. Infect Immun. 1983 Oct;42(1):250–256. doi: 10.1128/iai.42.1.250-256.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carof M., Lebbar S., Szabó L. Detection of 3-deoxy-2-octulosonic acid in thiobarbiturate-negative endotoxins. Carbohydr Res. 1987 Mar 15;161(1):c4–c7. doi: 10.1016/0008-6215(87)84019-3. [DOI] [PubMed] [Google Scholar]
  4. Caroff M., Deprun C., Richards J. C., Karibian D. Structural characterization of the lipid A of Bordetella pertussis 1414 endotoxin. J Bacteriol. 1994 Aug;176(16):5156–5159. doi: 10.1128/jb.176.16.5156-5159.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Di Padova F. E., Brade H., Barclay G. R., Poxton I. R., Liehl E., Schuetze E., Kocher H. P., Ramsay G., Schreier M. H., McClelland D. B. A broadly cross-protective monoclonal antibody binding to Escherichia coli and Salmonella lipopolysaccharides. Infect Immun. 1993 Sep;61(9):3863–3872. doi: 10.1128/iai.61.9.3863-3872.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edebrink P., Jansson P. E., Rahman M. M., Widmalm G., Holme T., Rahman M., Weintraub A. Structural studies of the O-polysaccharide from the lipopolysaccharide of Moraxella (Branhamella) catarrhalis serotype A (strain ATCC 25238). Carbohydr Res. 1994 May 5;257(2):269–284. doi: 10.1016/0008-6215(94)80040-5. [DOI] [PubMed] [Google Scholar]
  7. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  8. Gibson B. W., Melaugh W., Phillips N. J., Apicella M. A., Campagnari A. A., Griffiss J. M. Investigation of the structural heterogeneity of lipooligosaccharides from pathogenic Haemophilus and Neisseria species and of R-type lipopolysaccharides from Salmonella typhimurium by electrospray mass spectrometry. J Bacteriol. 1993 May;175(9):2702–2712. doi: 10.1128/jb.175.9.2702-2712.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Helander I. M., Lindner B., Brade H., Altmann K., Lindberg A. A., Rietschel E. T., Zähringer U. Chemical structure of the lipopolysaccharide of Haemophilus influenzae strain I-69 Rd-/b+. Description of a novel deep-rough chemotype. Eur J Biochem. 1988 Nov 15;177(3):483–492. doi: 10.1111/j.1432-1033.1988.tb14398.x. [DOI] [PubMed] [Google Scholar]
  10. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hitchcock P. J., Leive L., Mäkelä P. H., Rietschel E. T., Strittmatter W., Morrison D. C. Lipopolysaccharide nomenclature--past, present, and future. J Bacteriol. 1986 Jun;166(3):699–705. doi: 10.1128/jb.166.3.699-705.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoffman W. D., Natanson C. Endotoxin in septic shock. Anesth Analg. 1993 Sep;77(3):613–624. doi: 10.1213/00000539-199309000-00032. [DOI] [PubMed] [Google Scholar]
  13. Kimura A., Hansen E. J. Antigenic and phenotypic variations of Haemophilus influenzae type b lipopolysaccharide and their relationship to virulence. Infect Immun. 1986 Jan;51(1):69–79. doi: 10.1128/iai.51.1.69-79.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuhn H. M. Cross-reactivity of monoclonal antibodies and sera directed against lipid A and lipopolysaccharides. Infection. 1993 May-Jun;21(3):179–186. doi: 10.1007/BF01710544. [DOI] [PubMed] [Google Scholar]
  15. Lebbar S., Caroff M., Szabó L., Mérienne C., Szilógyi L. Structure of a hexasaccharide proximal to the hydrophobic region of lipopolysaccharides present in Bordetella pertussis endotoxin preparations. Carbohydr Res. 1994 Jun 17;259(2):257–275. doi: 10.1016/0008-6215(94)84061-x. [DOI] [PubMed] [Google Scholar]
  16. Maskell D. J., Szabo M. J., Butler P. D., Williams A. E., Moxon E. R. Molecular analysis of a complex locus from Haemophilus influenzae involved in phase-variable lipopolysaccharide biosynthesis. Mol Microbiol. 1991 May;5(5):1013–1022. doi: 10.1111/j.1365-2958.1991.tb01874.x. [DOI] [PubMed] [Google Scholar]
  17. Maskell D. J., Szabo M. J., Butler P. D., Williams A. E., Moxon E. R. Phase variation of lipopolysaccharide in Haemophilus influenzae. Res Microbiol. 1991 Jul-Aug;142(6):719–724. doi: 10.1016/0923-2508(91)90086-p. [DOI] [PubMed] [Google Scholar]
  18. Maskell D. J., Szabo M. J., Deadman M. E., Moxon E. R. The gal locus from Haemophilus influenzae: cloning, sequencing and the use of gal mutants to study lipopolysaccharide. Mol Microbiol. 1992 Oct;6(20):3051–3063. doi: 10.1111/j.1365-2958.1992.tb01763.x. [DOI] [PubMed] [Google Scholar]
  19. Melaugh W., Phillips N. J., Campagnari A. A., Karalus R., Gibson B. W. Partial characterization of the major lipooligosaccharide from a strain of Haemophilus ducreyi, the causative agent of chancroid, a genital ulcer disease. J Biol Chem. 1992 Jul 5;267(19):13434–13439. [PubMed] [Google Scholar]
  20. Michon F., Shaw D. H., Banoub J. H. Structure of the lipopolysaccharide core isolated from a human strain of Aeromonas hydrophila. Eur J Biochem. 1984 Nov 15;145(1):107–114. doi: 10.1111/j.1432-1033.1984.tb08528.x. [DOI] [PubMed] [Google Scholar]
  21. Nnalue N. A., Lind S. M., Lindberg A. A. The disaccharide L-alpha-D-heptose1-->7-L-alpha-D-heptose1-->of the inner core domain of Salmonella lipopolysaccharide is accessible to antibody and is the epitope of a broadly reactive monoclonal antibody. J Immunol. 1992 Oct 15;149(8):2722–2728. [PubMed] [Google Scholar]
  22. Phillips N. J., Apicella M. A., Griffiss J. M., Gibson B. W. Structural characterization of the cell surface lipooligosaccharides from a nontypable strain of Haemophilus influenzae. Biochemistry. 1992 May 12;31(18):4515–4526. doi: 10.1021/bi00133a019. [DOI] [PubMed] [Google Scholar]
  23. Phillips N. J., Apicella M. A., Griffiss J. M., Gibson B. W. Structural studies of the lipooligosaccharides from Haemophilus influenzae type b strain A2. Biochemistry. 1993 Mar 2;32(8):2003–2012. doi: 10.1021/bi00059a017. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Qureshi N., Takayama K., Ribi E. Purification and structural determination of nontoxic lipid A obtained from the lipopolysaccharide of Salmonella typhimurium. J Biol Chem. 1982 Oct 10;257(19):11808–11815. [PubMed] [Google Scholar]
  26. Schweda E. K., Hegedus O. E., Borrelli S., Lindberg A. A., Weiser J. N., Maskell D. J., Moxon E. R. Structural studies of the saccharide part of the cell envelope lipopolysaccharide from Haemophilus influenzae strain AH1-3 (lic3+). Carbohydr Res. 1993 Aug 17;246:319–330. doi: 10.1016/0008-6215(93)84043-6. [DOI] [PubMed] [Google Scholar]
  27. Schweda E. K., Sundström A. C., Eriksson L. M., Jonasson J. A., Lindberg A. A. Structural studies of the cell envelope lipopolysaccharides from Haemophilus ducreyi strains ITM 2665 and ITM 4747. J Biol Chem. 1994 Apr 22;269(16):12040–12048. [PubMed] [Google Scholar]
  28. Shaw D. H., Hart M. J., Lüderitz O. Structure of the core oligosaccharide in the lipopolysaccharide isolated from Aeromonas salmonicida ssp. salmonicida. Carbohydr Res. 1992 Jul 2;231:83–91. doi: 10.1016/0008-6215(92)84010-p. [DOI] [PubMed] [Google Scholar]
  29. Shaw D. H., Squires M. J., Ishiguro E. E., Trust T. J. The structure of the heptose-3-deoxy-D-mannooctulosonic-acid region in a mutant form of Aeromonas salmonicida lipopolysaccharide. Eur J Biochem. 1986 Dec 1;161(2):309–313. doi: 10.1111/j.1432-1033.1986.tb10448.x. [DOI] [PubMed] [Google Scholar]
  30. Spinola S. M., Kwaik Y. A., Lesse A. J., Campagnari A. A., Apicella M. A. Cloning and expression in Escherichia coli of a Haemophilus influenzae type b lipooligosaccharide synthesis gene(s) that encodes a 2-keto-3-deoxyoctulosonic acid epitope. Infect Immun. 1990 Jun;58(6):1558–1564. doi: 10.1128/iai.58.6.1558-1564.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Szabó P., Szabó L. Synthesis of 2-deoxy-4-O-alpha- and -beta-D-mannopyranosyl-L-erythro-pentonic acids. Carbohydr Res. 1991 Sep 2;216:227–235. doi: 10.1016/0008-6215(92)84164-n. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Verheul A. F., Snippe H., Poolman J. T. Meningococcal lipopolysaccharides: virulence factor and potential vaccine component. Microbiol Rev. 1993 Mar;57(1):34–49. doi: 10.1128/mr.57.1.34-49.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Virji M., Weiser J. N., Lindberg A. A., Moxon E. R. Antigenic similarities in lipopolysaccharides of Haemophilus and Neisseria and expression of a digalactoside structure also present on human cells. Microb Pathog. 1990 Dec;9(6):441–450. doi: 10.1016/0882-4010(90)90062-u. [DOI] [PubMed] [Google Scholar]
  35. Weiser J. N., Lindberg A. A., Manning E. J., Hansen E. J., Moxon E. R. Identification of a chromosomal locus for expression of lipopolysaccharide epitopes in Haemophilus influenzae. Infect Immun. 1989 Oct;57(10):3045–3052. doi: 10.1128/iai.57.10.3045-3052.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weiser J. N., Maskell D. J., Butler P. D., Lindberg A. A., Moxon E. R. Characterization of repetitive sequences controlling phase variation of Haemophilus influenzae lipopolysaccharide. J Bacteriol. 1990 Jun;172(6):3304–3309. doi: 10.1128/jb.172.6.3304-3309.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiser J. N., Williams A., Moxon E. R. Phase-variable lipopolysaccharide structures enhance the invasive capacity of Haemophilus influenzae. Infect Immun. 1990 Oct;58(10):3455–3457. doi: 10.1128/iai.58.10.3455-3457.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wenger J. D. Impact of Haemophilus influenzae type b vaccines on the epidemiology of bacterial meningitis. Infect Agents Dis. 1993 Oct;2(5):324–332. [PubMed] [Google Scholar]
  39. 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]
  40. Zwahlen A., Rubin L. G., Connelly C. J., Inzana T. J., Moxon E. R. Alteration of the cell wall of Haemophilus influenzae type b by transformation with cloned DNA: association with attenuated virulence. J Infect Dis. 1985 Sep;152(3):485–492. doi: 10.1093/infdis/152.3.485. [DOI] [PubMed] [Google Scholar]
  41. de StGroth S. F., Scheidegger D. Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 1980;35(1-2):1–21. doi: 10.1016/0022-1759(80)90146-5. [DOI] [PubMed] [Google Scholar]

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