Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 Mar;60(3):1069–1075. doi: 10.1128/iai.60.3.1069-1075.1992

Modulation of effects of lipopolysaccharide on macrophages by a major outer membrane protein of Proteus mirabilis as measured in a chemiluminescence assay.

G Weber 1, D Heck 1, R R Bartlett 1, K Nixdorff 1
PMCID: PMC257595  PMID: 1541521

Abstract

Our previous studies have shown that a major protein isolated from purified cell walls of Proteus mirabilis (39-kDa protein) is a strong modulator of the specific immune responses to lipopolysaccharide (LPS) from this bacterium. When the protein is mixed with LPS before immunization of mice, the responses of antibody-producing cells specific for LPS are greatly enhanced and converted predominantly to the immunoglobulin G isotype. In the present study, the immunomodulating effects of the 39-kDa protein were tested at the level of interaction of LPS with macrophages. Activation of macrophages was determined by measuring the production of oxygen radicals in a chemiluminescence assay with lucigenin as the amplifier. LPS from P. mirabilis induced strong oxidative metabolism in both peritoneal and bone marrow-derived murine macrophages. These responses were inhibited in a dose-dependent manner by mixing LPS with increasing amounts of the protein. In contrast, bovine serum albumin and methylated bovine serum albumin enhanced the response of macrophages dramatically when complexed with LPS. The inhibiting activity of the 39-kDa protein was also observed with LPS from Escherichia coli K-12.

Full text

PDF
1069

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aderem A. A., Cohen D. S., Wright S. D., Cohn Z. A. Bacterial lipopolysaccharides prime macrophages for enhanced release of arachidonic acid metabolites. J Exp Med. 1986 Jul 1;164(1):165–179. doi: 10.1084/jem.164.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aderem A. A., Cohn Z. A. Calcium ionophore synergizes with bacterial lipopolysaccharides in activating macrophage arachidonic acid metabolism. J Exp Med. 1988 Feb 1;167(2):623–631. doi: 10.1084/jem.167.2.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Flesch I., Ecker B., Ferber E. Acyltransferase-catalyzed cleavage of arachidonic acid from phospholipids and transfer to lysophosphatides in macrophages derived from bone marrow. Comparison of different donor- and acceptor substrate combinations. Eur J Biochem. 1984 Mar 15;139(3):431–437. doi: 10.1111/j.1432-1033.1984.tb08023.x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Galanos C., Rietschel E. T., Lüderitz O., Westphal O., Kim Y. B., Watson D. W. Biological activities of lipid A complexed with bovine-serum albumin. Eur J Biochem. 1972 Dec 4;31(2):230–233. doi: 10.1111/j.1432-1033.1972.tb02524.x. [DOI] [PubMed] [Google Scholar]
  6. Gmeiner J. The isolation of two different lipopolysaccharide fractions from various Proteus mirabilis strains. Eur J Biochem. 1975 Oct 15;58(2):621–626. doi: 10.1111/j.1432-1033.1975.tb02413.x. [DOI] [PubMed] [Google Scholar]
  7. Kapp A., Freudenberg M., Galanos C. Induction of human granulocyte chemiluminescence by bacterial lipopolysaccharides. Infect Immun. 1987 Mar;55(3):758–761. doi: 10.1128/iai.55.3.758-761.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Karch H., Gmeiner J., Nixdorff K. Alteration of the immunoglobulin G subclass responses in mice to lipopolysaccharide: effects of nonbacterial proteins and bacterial membrane phospholipids or outer membrane proteins of Proteus mirabilis. Infect Immun. 1983 Apr;40(1):157–165. doi: 10.1128/iai.40.1.157-165.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Karch H., Nixdorff K. Antibody-producing cell responses to an isolated outer membrane protein and to complexes of this antigen with lipopolysaccharide or with vesicles of phospholipids from Proteus mirabilis. Infect Immun. 1981 Mar;31(3):862–867. doi: 10.1128/iai.31.3.862-867.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Karch H., Nixdorff K. Modulation of the IgG subclass responses to lipopolysaccharide by bacterial membrane components: differential adjuvant effects produced by primary and secondary stimulation. J Immunol. 1983 Jul;131(1):6–8. [PubMed] [Google Scholar]
  11. Müller-Peddinghaus R. In vitro determination of phagocyte activity by luminol- and lucigenin-amplified chemiluminescence. Int J Immunopharmacol. 1984;6(5):455–466. doi: 10.1016/0192-0561(84)90084-5. [DOI] [PubMed] [Google Scholar]
  12. Nixdorff K., Fitzer H., Gmeiner J., Martin H. H. Reconstitution of model membranes from phospholipid and outer membrane proteins of Proteus mirabilis. Role of proteins in the formation of hydrophilic pores and protection of membranes against detergents. Eur J Biochem. 1977 Nov 15;81(1):63–69. doi: 10.1111/j.1432-1033.1977.tb11927.x. [DOI] [PubMed] [Google Scholar]
  13. Schumann R. R., Leong S. R., Flaggs G. W., Gray P. W., Wright S. D., Mathison J. C., Tobias P. S., Ulevitch R. J. Structure and function of lipopolysaccharide binding protein. Science. 1990 Sep 21;249(4975):1429–1431. doi: 10.1126/science.2402637. [DOI] [PubMed] [Google Scholar]
  14. Tobias P. S., Soldau K., Ulevitch R. J. Isolation of a lipopolysaccharide-binding acute phase reactant from rabbit serum. J Exp Med. 1986 Sep 1;164(3):777–793. doi: 10.1084/jem.164.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wright S. D. Multiple receptors for endotoxin. Curr Opin Immunol. 1991 Feb;3(1):83–90. doi: 10.1016/0952-7915(91)90082-c. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES