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Structural and Immunochemical Aspects of Brucella Abortus Endotoxins

  • Albert M. Wu
  • Neil E. MacKenzie
  • L. Garry Adams
  • Roberta Pugh
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 228)

Abstract

The smooth lipopolysaccharide (sLPS)-protein complex of Brucella abortus has been identified as the immuno-dominant component, which reacts with antibody in the serological tests generally used for diagnosis of brucellosis (1). B. abortus sLPS carries the major A (abortus) antigen, the species-specific epitope located within, and the minor M (melitensis) determinant, which cross reacts with B. melitensis antigen. O-chain prepared fromB. abortus LPS was reported to have protective antigen properties in mice (2). Unlike the LPS of other enterobacteriaceae, sLPS separates primarily into the phenol phase when B. abortus cells are extracted with hot phenol-water by the Westphal method (3–7). The crude fraction of sLPS, which was used to study the properties of smooth B. abortus in the early eighties, is associated with various proteins ranging from 10–30% of the weight composition, most of which are non-covalently bound (3). These non-covalently linked proteins or nucleic acids can be removed by treatment with chaotropic agents and by gel filtration in the presence of chaotropic agents and detergents.

Keywords

Phenol Sulfuric Acid Brucella Abortus Brucella Melitensis Chaotropic Agent Phenol Phase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations Used

LPS

lipopolysaccharides

sLPS

smooth lipopoly-saccharides

cLPS

crude lipopolysaccharides

AH

acid hapten

KDO

3-deoxyoctulosonic acid

ELISA

enzyme-linked immunosorbent assay

LAL

limulus amoebocyte lysate test

HexN

Hexosamine

PS

phenol sulfuric acid method

O.

orcinol method

Glc

DGlucopyranose

Gal

DGalactopyranose

Man

DMannopyranose

MeaMan

MethylDMannopyranoside

MWCO

Molecular weight cut off

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References

  1. 1.
    Berman, D.T. (1981) The importance of being earnest with your antigens. In: The Ruminant Immune System (Edited by Butler, J.E.), pp. 217–224. Plenum Press, New York and London.Google Scholar
  2. 2.
    Montaraz, J.A., Winter, A.J., Hunter, D.M., Sowa, B.A., Wu, A.M. and Adams, L.G.(1986) Protection against Brucella abortus in mice and O-polysaccharide specific monoclonal antibodies. Infect. Immun., 51:961–963.PubMedGoogle Scholar
  3. 3.
    Moreno, E., Pitt, M.W., Jones, L.M., Schurig, G.G. and Berman, D.T. (1979). Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus. J. Bacteriol., 138:361–369.PubMedGoogle Scholar
  4. 4.
    Baker, P. and Wilson, J.B. (1965). Hypoferremia in mice and its application of the bioassay of endotoxin. J. Bacteriol., 90:903–910.PubMedGoogle Scholar
  5. 5.
    Marx, A., Jonescu, J. and Pop, A. (1983). Immunochemical studies on Brucella abortus lipopolysaccharide. Zbl. Bakt. Hyg. I Abt. Orig. A 253:544–553.Google Scholar
  6. 6.
    Caroff, M., Bundle, D.R., Perry, M.B., Cherwonogrodzky, J.W. and Duncan J.R. (1984).Antigenic S-type lipopolysaccharide of Brucella abortus S1119–3. Infect, and Immun., 46:384–388.Google Scholar
  7. 7.
    Wu, A.M., Heck, F.C., Adams, L.G. and Jones, K. (1984) Immunochemical studies on the binding properties of Brucella abortus lipopolysaccharides to bovine precipitating antibodies. Mol. Immun., 21:1123–1129.CrossRefGoogle Scholar
  8. 8.
    Perera, V.Y., Winter, A.J., and Ganem, B. (1984) Evidence for covalent bondings of native hapten protein complexes to smooth lipopolysaccharide of Brucella abortus. FEMS Lett., 211:263–266.CrossRefGoogle Scholar
  9. 9.
    Wu, A.M., Adams L.G. and Pugh, R. (1987) Immunochemical and partial chemical characterization of fractions of membrane-bound smooth lipopolysaccharide-protein complexes from Brucella abortus. Mol. Cell. Biochem., 75:93–102.PubMedGoogle Scholar
  10. 10.
    Wu, A.M. and Maenzie, N.E. (1987) Structural and immunochemical characterization of the O-haptens of Brucella abortus lipopolysaccharides from strains 19 and 2308. Mol. Cell. Biochem., 75:103–111.PubMedGoogle Scholar
  11. 11.
    Caroff, M., Bundle, D.R. and Perry, M.B. (1984) Structure of the O-chain of the phenol-phase soluble cellular lipopolysaccharide of Yersinia enterocolitica serotype 0:9. Eur. J. Biochem. 139:195–200.PubMedCrossRefGoogle Scholar
  12. 12.
    Redmond, J.W. (1978) The 4-amino sugars present in the lipopolysaccharides of Vibrio cholerae and related Vibrios. Biochim. Biophys. Acta., 542:378–384.PubMedGoogle Scholar
  13. 13.
    Moreno, E., Berman, D.T., and Boettcher, L. A. (1981) Biological activities of Brucella abortus lipopolysaccharides. Infec. Immun., 31:362–370.Google Scholar
  14. 14.
    Moreno, E., Speth, S.L., Jones, L.J., and Berman, D.T. (1981) Immunochemical characterization of Brucella lipopolysaccharides and polysaccharides. Infec. Immun., 31: 214–222.Google Scholar
  15. 15.
    Tsai, C.M. and Frasch, C.E. (1982) A sensitive silver stain for detecting lipopolysaccharides in Polyacrylamide gels. Analyt. Biochem. 119:115–119.PubMedCrossRefGoogle Scholar
  16. 16.
    Jann, B., Reske, K. and Jann, K., (1975) Heterogeneity of Lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Eur. J. Biochem. 60:239–246.PubMedCrossRefGoogle Scholar
  17. 17.
    Goldman, R.C., and Leive, L. (1980) Heterogeneity of antigenic side chain length in lipopolysaccharide from Escherichia 0111 and Salmonella typhimurium LT2. Eur. J. Biochem. 107:145–153.PubMedCrossRefGoogle Scholar
  18. 18.
    Palva, E.T. and Mäkelä (1980) Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate Polyacrylamide gel electrophoresis. Eur. J. Biochem. 107:137–143.PubMedCrossRefGoogle Scholar
  19. 19.
    Bundle, D.R., Perry, M.B., and Cherwonogrodzky, J.W. (1987) Structural elucidation of Brucella melitensis M antigen by high-resolution NMR at 500MHz, Biochemistry 26:8717–8726.PubMedCrossRefGoogle Scholar
  20. 19A.
    Moreno, E., Mayer, H., and Moriyon, 1.(1987) Characterization of a native polysaccharide hapten from Brucella melitensis. Infec. Immun. 55:2850–2853.Google Scholar
  21. 20.
    Bundle, D.R., Cherwonogrodzky, J.W., Caroff, M. and Perry, M.B. (1987) The Lipopolysaccharide of Brucella abortus and B. Melitensis. Ann. Inst. Pasteur./Microbiol. 138:92–98.CrossRefGoogle Scholar
  22. 21.
    Maclntyre, S., Lucken, R., and Own, P. (1986) Smooth lipopolysaccharide is the major protective antigen for mice in the surface extract from IATS serotype 6 contributing to the polyvalent Pseudomonas aeruginosa vaccine PEV. Infec. Immun., 52:76–84.Google Scholar
  23. 22.
    Cryz, S.J., Jr., Furer, E., and Germanier, R. (1984) Protection against fatal Pseudomonas aeruginosa burn wound sepsis by immunization with lipopolysaccharide and high-molecular weight polysaccharide. Infect. Immun., 43:795–799.PubMedGoogle Scholar
  24. 23.
    Pier, G.B. (1983) Immunochemistry of Pseudomonas aeruginosa lipopolysaccharides and high-molecular-weight polysaccharides. Rev. Infect. Dis. 5 (Suppl.):S950-S956.PubMedCrossRefGoogle Scholar
  25. 24.
    Pollach, M., Pier, GG., and Prescott, R.K. (1984) Immunization with Pseudomonas aeruginosa high-molecular-weight polysaccharides prevent death from Pseudomonas burn infections in mice. Infect. Immun., 43:759–760.Google Scholar
  26. Cryz, S.J., Furer, E., Sadoff, J.C., and Germanier, R. (1986) Pseudomonas aeruginosa immunotypes polysaccharide-Toxin A conjugate vaccine. Infect. Immun., 161–165.Google Scholar
  27. 26.
    Seid, R.C., and Sadoff, J.C. (1981) Preparation and characterization of detoxified lipopolysaccharide-protein conjugates. J. Biol. Chem., 256:7305–7310.PubMedGoogle Scholar
  28. 27.
    Enlanger, B.F., The preparation of antigenic hapten-carrier conjugates: A survey. (1980) Meth. Enzymol., 70:85–104.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Albert M. Wu
    • 1
  • Neil E. MacKenzie
    • 2
    • 3
  • L. Garry Adams
    • 1
  • Roberta Pugh
    • 2
  1. 1.Department of Veterinary PathologyCollege of Veterinary MedicineCollege StationUSA
  2. 2.Department of Veterinary Microbiology and ParasitologyTexas A&M UniversityCollege StationUSA
  3. 3.Department of Pharmaceutic Science, College of PharmacyUniversity of ArizonaTucsonUSA

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