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. 1997 Sep;65(9):3961–3965. doi: 10.1128/iai.65.9.3961-3965.1997

A nonsubstituted primary hydroxyl group in position 6' of free lipid A is required for binding of lipid A monoclonal antibodies.

L Brade 1, R Engel 1, W J Christ 1, E T Rietschel 1
PMCID: PMC175568  PMID: 9284181

Abstract

Lipid A monoclonal antibodies, which require for binding the presence of the bisphosphorylated D-glucosamine disaccharide lipid A backbone, were tested against synthetic lipid A precursor Ia and compound B 1047 by enzyme immunoassay. The last-named compound is a precursor Ia analog with a methoxy instead of a hydroxy group at C6' and was chosen to determine why these antibodies failed to recognize the bound lipid A present in lipopolysaccharide (LPS). Whereas all antibodies tested bound to precursor Ia, none of them bound to compound B 1047 or Escherichia coli Re LPS to a significant extent. Compared to the natural substituent at C6', i.e., 3-deoxy-D-manno-octulosonic acid (Kdo), the methoxy group is neither bulky nor charged. Thus, the data suggest that it is not hindrance by Kdo but rather the generation of a neoantigen that endows lipid A with immunoreactivity upon liberation from LPS by acid hydrolysis.

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Selected References

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  1. Bogard W. C., Jr, Dunn D. L., Abernethy K., Kilgarriff C., Kung P. C. Isolation and characterization of murine monoclonal antibodies specific for gram-negative bacterial lipopolysaccharide: association of cross-genus reactivity with lipid A specificity. Infect Immun. 1987 Apr;55(4):899–908. doi: 10.1128/iai.55.4.899-908.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brade L., Brade H. Characterization of two different antibody specificities recognizing distinct antigenic determinants in free lipid A of Escherichia coli. Infect Immun. 1985 Jun;48(3):776–781. doi: 10.1128/iai.48.3.776-781.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brade L., Brandenburg K., Kuhn H. M., Kusumoto S., Macher I., Rietschel E. T., Brade H. The immunogenicity and antigenicity of lipid A are influenced by its physicochemical state and environment. Infect Immun. 1987 Nov;55(11):2636–2644. doi: 10.1128/iai.55.11.2636-2644.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brade L., Holst O., Brade H. An artificial glycoconjugate containing the bisphosphorylated glucosamine disaccharide backbone of lipid A binds lipid A monoclonal antibodies. Infect Immun. 1993 Oct;61(10):4514–4517. doi: 10.1128/iai.61.10.4514-4517.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brade L., Rietschel E. T., Kusumoto S., Shiba T., Brade H. Immunogenicity and antigenicity of synthetic Escherichia coli lipid A. Infect Immun. 1986 Jan;51(1):110–114. doi: 10.1128/iai.51.1.110-114.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christ W. J., Asano O., Robidoux A. L., Perez M., Wang Y., Dubuc G. R., Gavin W. E., Hawkins L. D., McGuinness P. D., Mullarkey M. A. E5531, a pure endotoxin antagonist of high potency. Science. 1995 Apr 7;268(5207):80–83. doi: 10.1126/science.7701344. [DOI] [PubMed] [Google Scholar]
  7. Dunn D. L., Ewald D. C., Chandan N., Cerra F. B. Immunotherapy of gram-negative bacterial sepsis. A single murine monoclonal antibody provides cross-genera protection. Arch Surg. 1986 Jan;121(1):58–62. doi: 10.1001/archsurg.1986.01400010064008. [DOI] [PubMed] [Google Scholar]
  8. Flad H. D., Loppnow H., Rietschel E. T., Ulmer A. J. Agonists and antagonists for lipopolysaccharide-induced cytokines. Immunobiology. 1993 Apr;187(3-5):303–316. doi: 10.1016/S0171-2985(11)80346-3. [DOI] [PubMed] [Google Scholar]
  9. Fujihara Y., Lei M. G., Morrison D. C. Characterization of specific binding of a human immunoglobulin M monoclonal antibody to lipopolysaccharide and its lipid A domain. Infect Immun. 1993 Mar;61(3):910–918. doi: 10.1128/iai.61.3.910-918.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Galanos C., Freudenberg M. A., Jay F., Nerkar D., Veleva K., Brade H., Strittmatter W. Immunogenic properties of lipid A. Rev Infect Dis. 1984 Jul-Aug;6(4):546–552. doi: 10.1093/clinids/6.4.546. [DOI] [PubMed] [Google Scholar]
  11. Galanos C., Lehmann V., Lüderitz O., Rietschel E. T., Westphal O., Brade H., Brade L., Freudenberg M. A., Hansen-Hagge T., Lüderitz T. Endotoxic properties of chemically synthesized lipid A part structures. Comparison of synthetic lipid A precursor and synthetic analogues with biosynthetic lipid A precursor and free lipid A. Eur J Biochem. 1984 Apr 16;140(2):221–227. doi: 10.1111/j.1432-1033.1984.tb08090.x. [DOI] [PubMed] [Google Scholar]
  12. Galanos C., Lüderitz O., Rietschel E. T., Westphal O., Brade H., Brade L., Freudenberg M., Schade U., Imoto M., Yoshimura H. Synthetic and natural Escherichia coli free lipid A express identical endotoxic activities. Eur J Biochem. 1985 Apr 1;148(1):1–5. doi: 10.1111/j.1432-1033.1985.tb08798.x. [DOI] [PubMed] [Google Scholar]
  13. Kuhn H. M., Brade L., Appelmelk B. J., Kusumoto S., Rietschel E. T., Brade H. Characterization of the epitope specificity of murine monoclonal antibodies directed against lipid A. Infect Immun. 1992 Jun;60(6):2201–2210. doi: 10.1128/iai.60.6.2201-2210.1992. [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. Lynn W. A., Golenbock D. T. Lipopolysaccharide antagonists. Immunol Today. 1992 Jul;13(7):271–276. doi: 10.1016/0167-5699(92)90009-V. [DOI] [PubMed] [Google Scholar]
  16. Pollack M., Ohl C. A. Endotoxin-based molecular strategies for the prevention and treatment of gram-negative sepsis and septic shock. Curr Top Microbiol Immunol. 1996;216:275–297. doi: 10.1007/978-3-642-80186-0_13. [DOI] [PubMed] [Google Scholar]
  17. Pollack M., Raubitschek A. A., Larrick J. W. Human monoclonal antibodies that recognize conserved epitopes in the core-lipid A region of lipopolysaccharides. J Clin Invest. 1987 May;79(5):1421–1430. doi: 10.1172/JCI112970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rietschel E. T., Brade H., Holst O., Brade L., Müller-Loennies S., Mamat U., Zähringer U., Beckmann F., Seydel U., Brandenburg K. Bacterial endotoxin: Chemical constitution, biological recognition, host response, and immunological detoxification. Curr Top Microbiol Immunol. 1996;216:39–81. doi: 10.1007/978-3-642-80186-0_3. [DOI] [PubMed] [Google Scholar]
  19. Rietschel E. T., Galanos C. Lipid A antiserum-mediated protection against lipopolysaccharide- and lipid A-induced fever and skin necrosis. Infect Immun. 1977 Jan;15(1):34–49. doi: 10.1128/iai.15.1.34-49.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rietschel E. T., Kirikae T., Schade F. U., Mamat U., Schmidt G., Loppnow H., Ulmer A. J., Zähringer U., Seydel U., Di Padova F. Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB J. 1994 Feb;8(2):217–225. doi: 10.1096/fasebj.8.2.8119492. [DOI] [PubMed] [Google Scholar]
  21. Rozalski A., Brade L., Kuhn H. M., Brade H., Kosma P., Appelmelk B. J., Kusumoto S., Paulsen H. Determination of the epitope specificity of monoclonal antibodies against the inner core region of bacterial lipopolysaccharides by use of 3-deoxy-D-manno-octulosonate-containing synthetic antigens. Carbohydr Res. 1989 Oct 31;193:257–270. doi: 10.1016/0008-6215(89)85124-9. [DOI] [PubMed] [Google Scholar]
  22. Zähringer U., Lindner B., Rietschel E. T. Molecular structure of lipid A, the endotoxic center of bacterial lipopolysaccharides. Adv Carbohydr Chem Biochem. 1994;50:211–276. [PubMed] [Google Scholar]

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