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. 1994 Mar;62(3):1052–1057. doi: 10.1128/iai.62.3.1052-1057.1994

Binding studies of a monoclonal antibody specific for 3-deoxy-D-manno-octulosonic acid with a panel of Klebsiella pneumoniae lipopolysaccharides representing all of the O serotypes.

N M van der Meer 1, B J Appelmelk 1, A M Verweij-van Vught 1, W Nimmich 1, P Kosma 1, L G Thijs 1, J de Graaff 1, D M MacLaren 1
PMCID: PMC186223  PMID: 8112839

Abstract

A monoclonal antibody (MAb) raised against Salmonella minnesota R595 and specific for alpha-3-deoxy-D-manno-octulosonic acid (alpha-Kdo) of the inner core was tested for binding to lipopolysaccharides (LPS) of Klebsiella pneumoniae. The MAb was tested in several assay systems (enzyme-linked immunosorbent assay, passive hemolysis, and inhibition of passive hemolysis) with a large panel (n = 23) of K. pneumoniae LPS representing all nine currently known O serotypes. MAb 20 showed reactivity with almost all O serotypes of K. pneumoniae LPS, and this reactivity could be inhibited by synthetic Kdo. This suggests an epitope in the cores of these Klebsiella LPS much like that in the inner core of LPS of S. minnesota. Large differences in reactivity between LPS of different strains belonging to the same O serotype were observed. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis of LPS followed by immunoblotting, reactivity of MAb 20 was observed only with the fast-moving fraction possibly representing the nonsubstituted core. No binding was seen with the high-molecular-weight fraction that contained core material substituted with several units of O-antigen building blocks. The chemical basis for these differences in reactivity remains to be established. As far as we know, this is the first report containing comprehensive immunochemical data on the LPS core of K. pneumoniae.

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Selected References

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  1. Appelmelk B. J., Verweij-van Vught A. M., Brade H., Maaskant J. J., Schouten W. F., Thijs L. G., MacLaren D. M. Prevention of lethal endotoxemia in actinomycin D-sensitized mice by incubation of Salmonella minnesota R595 lipopolysaccharide with monoclonal antibodies to R595. Microb Pathog. 1988 Oct;5(4):251–257. doi: 10.1016/0882-4010(88)90097-6. [DOI] [PubMed] [Google Scholar]
  2. Baumgartner J. D. Immunotherapy with antibodies to core lipopolysaccharide: a critical appraisal. Infect Dis Clin North Am. 1991 Dec;5(4):915–927. [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Brade L., Kosma P., Appelmelk B. J., Paulsen H., Brade H. Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide. Infect Immun. 1987 Feb;55(2):462–466. doi: 10.1128/iai.55.2.462-466.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. DuPont H. L., Spink W. W. Infections due to gram-negative organisms: an analysis of 860 patients with bacteremia at the University of Minnesota Medical Center, 1958-1966. Medicine (Baltimore) 1969 Jul;48(4):307–332. doi: 10.1097/00005792-196907000-00003. [DOI] [PubMed] [Google Scholar]
  8. Erich T., Schellekens J., Bouter A., Van Kranen J., Brouwer E., Verhoef J. Binding characteristics and cross-reactivity of three different antilipid A monoclonal antibodies. J Immunol. 1989 Dec 15;143(12):4053–4060. [PubMed] [Google Scholar]
  9. Hasegawa T., Ohta M., Nakashima I., Kato N., Morikawa K., Harada T., Okuyama T. Structure of the polysaccharide moiety of the Klebsiella O3 lipopolysaccharide isolated from culture supernatant of decapsulated mutant (Klebsiella O3:K1-). Chem Pharm Bull (Tokyo) 1985 Jan;33(1):333–339. doi: 10.1248/cpb.33.333. [DOI] [PubMed] [Google Scholar]
  10. Kreger B. E., Craven D. E., Carling P. C., McCabe W. R. Gram-negative bacteremia. III. Reassessment of etiology, epidemiology and ecology in 612 patients. Am J Med. 1980 Mar;68(3):332–343. doi: 10.1016/0002-9343(80)90101-1. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Lesse A. J., Campagnari A. A., Bittner W. E., Apicella M. A. Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods. 1990 Jan 24;126(1):109–117. doi: 10.1016/0022-1759(90)90018-q. [DOI] [PubMed] [Google Scholar]
  13. Mandine E., Salles M. F., Zalisz R., Guenounou M., Smets P. Murine monoclonal antibodies to Klebsiella pneumoniae protect against lethal endotoxemia and experimental infection with capsulated K. pneumoniae. Infect Immun. 1990 Sep;58(9):2828–2833. doi: 10.1128/iai.58.9.2828-2833.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McCallus D. E., Norcross N. L. Antibody specific for Escherichia coli J5 cross-reacts to various degrees with an Escherichia coli clinical isolate grown for different lengths of time. Infect Immun. 1987 May;55(5):1042–1046. doi: 10.1128/iai.55.5.1042-1046.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nelson J. W., Barclay G. R., Micklem L. R., Poxton I. R., Govan J. R. Production and characterisation of mouse monoclonal antibodies reactive with the lipopolysaccharide core of Pseudomonas aeruginosa. J Med Microbiol. 1992 May;36(5):358–365. doi: 10.1099/00222615-36-5-358. [DOI] [PubMed] [Google Scholar]
  16. Nimmich W. Isolierung und chemische Zusammensetzung des Klebsiella-Antigens O1. Zentralbl Bakteriol Orig. 1969;210(4):494–501. [PubMed] [Google Scholar]
  17. Nimmich W., Korten G. Die chemische Zusammensetzung der Klebsiella-Lipopolysaccharide (O-Antigene) Pathol Microbiol (Basel) 1970;36(3):179–190. [PubMed] [Google Scholar]
  18. Nimmich W. Zur Isolierung und qualitativen Bausteinanalyse der K-Antigene von Klebsiellen. Z Med Mikrobiol Immunol. 1968;154(2):117–131. [PubMed] [Google Scholar]
  19. Ohta M., Kido N., Hasegawa T., Ito H., Fujii Y., Arakawa Y., Komatsu T., Kato N. Contribution of the mannan O side-chains to the adjuvant action of lipopolysaccharides. Immunology. 1987 Apr;60(4):503–507. [PMC free article] [PubMed] [Google Scholar]
  20. Pollack M., Chia J. K., Koles N. L., Miller M., Guelde G. Specificity and cross-reactivity of monoclonal antibodies reactive with the core and lipid A regions of bacterial lipopolysaccharide. J Infect Dis. 1989 Feb;159(2):168–188. doi: 10.1093/infdis/159.2.168. [DOI] [PubMed] [Google Scholar]
  21. Rozalski A., Brade L., Kosma P., Appelmelk B. J., Krogmann C., Brade H. Epitope specificities of murine monoclonal and rabbit polyclonal antibodies against enterobacterial lipopolysaccharides of the Re chemotype. Infect Immun. 1989 Sep;57(9):2645–2652. doi: 10.1128/iai.57.9.2645-2652.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Ziegler E. J., Fisher C. J., Jr, Sprung C. L., Straube R. C., Sadoff J. C., Foulke G. E., Wortel C. H., Fink M. P., Dellinger R. P., Teng N. N. Treatment of gram-negative bacteremia and septic shock with HA-1A human monoclonal antibody against endotoxin. A randomized, double-blind, placebo-controlled trial. The HA-1A Sepsis Study Group. N Engl J Med. 1991 Feb 14;324(7):429–436. doi: 10.1056/NEJM199102143240701. [DOI] [PubMed] [Google Scholar]

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