Abstract
Murine monoclonal and rabbit polyclonal antibodies raised against the lipopolysaccharides (LPS) of Re mutants of Salmonella minnesota, Proteus mirabilis, and Escherichia coli were serologically characterized. Using natural Re LPS and natural and synthetic partial structures thereof, representing the 3-deoxy-D-manno-2-octulosonic acid (KDO) or lipid A region or both, the epitope specificities of four monoclonal antibodies were defined. Clones 20 (immunoglobulin M [IgM]) and 25 (IgG3) recognize a terminal alpha-pyranosidically linked KDO monosaccharide residue and the alpha-2,4-linked KDO disaccharide, respectively, as the immunodominant group. Therefore, these two antibodies are core antibodies which do not require the presence of lipid A constituents for binding. The minimal structure enabling binding of clone 17 (IgG2b) is a pseudotetrasaccharide of the sequence alpha-KDO-(2----4)-alpha-KDO-(2----6)-beta-glucosamine-(1----6)- glucosaminitol with two amide-linked 3-hydroxytetradecanoic acid residues. The smallest structure with which clone 22 (IgG3) reacted was de-O-acylated Re LPS. Therefore, clones 17 and 22 are LPS antibodies requiring both the lipid A and the KDO region for binding. Phosphoryl residues of the lipid A moiety in Re LPS are dispensable for the reaction with clone 17, whereas they are necessary for that with clone 22. These four different antibody types were also detected in polyclonal rabbit antisera and could be distinguished from each other by absorption experiments. It was found that type 20 and 25 antibodies either were not present or were present only in small amounts and that the majority of the antibodies were of types 17 and 22. From these data, we conclude that the immunodominant structures of Re LPS comprise both the KDO and lipid A domains.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brade H., Brade L., Rietschel E. T. Structure-activity relationships of bacterial lipopolysaccharides (endotoxins). Current and future aspects. Zentralbl Bakteriol Mikrobiol Hyg A. 1988 Apr;268(2):151–179. doi: 10.1016/s0176-6724(88)80001-4. [DOI] [PubMed] [Google Scholar]
- Brade H., Rietschel E. T. Alpha-2----4-interlinked 3-deoxy-D-manno-octulosonic acid disaccharide. A common constituent of enterobacterial lipopolysaccharides. Eur J Biochem. 1984 Dec 3;145(2):231–236. doi: 10.1111/j.1432-1033.1984.tb08543.x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Chernyak AYa, Levinsky A. B., Dmitriev B. A., Kochetkov N. K. A new type of carbohydrate-containing synthetic antigen: synthesis of carbohydrate-containing polyacrylamide copolymers having the specificity of O:3 and O:4 factors of Salmonella. Carbohydr Res. 1984 Jun 1;128(2):269–282. doi: 10.1016/0008-6215(84)85334-3. [DOI] [PubMed] [Google Scholar]
- Galanos C., Lüderitz O. Electrodialysis of lipopolysaccharides and their conversion to uniform salt forms. Eur J Biochem. 1975 Jun;54(2):603–610. doi: 10.1111/j.1432-1033.1975.tb04172.x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Kosma P., Gass J., Schulz G., Christian R., Unger F. M. Artificial antigens. Synthesis of polyacrylamide copolymers containing 3-deoxy-D-manno-2-octulopyranosylonic acid (KDO) residues. Carbohydr Res. 1987 Sep 15;167:39–54. doi: 10.1016/0008-6215(87)80266-5. [DOI] [PubMed] [Google Scholar]
- Lüderitz O., Galanos C., Risse H. J., Ruschmann E., Schlecht S., Schmidt G., Schulte-Holthausen H., Wheat R., Westphal O., Schlosshardt J. Structural relationship of Salmonella O and R antigens. Ann N Y Acad Sci. 1966 Jun 30;133(2):349–374. doi: 10.1111/j.1749-6632.1966.tb52376.x. [DOI] [PubMed] [Google Scholar]
- Lüderitz O., Staub A. M., Westphal O. Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae. Bacteriol Rev. 1966 Mar;30(1):192–255. doi: 10.1128/br.30.1.192-255.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sidorczyk Z., Kaca W., Brade H., Rietschel E. T., Sinnwell V., Zähringer U. Isolation and structural characterization of an 8-O-(4-amino-4-deoxy-beta-L-arabinopyranosyl)-3-deoxy-D-manno- octulosonic acid disaccharide in the lipopolysaccharide of a Proteus mirabilis deep rough mutant. Eur J Biochem. 1987 Oct 15;168(2):269–273. doi: 10.1111/j.1432-1033.1987.tb13416.x. [DOI] [PubMed] [Google Scholar]