Advertisement

Human complement activation by smooth and rough Proteus mirabilis lipopolysaccharides

  • Wiesław Kaca
  • Michał ArabskiEmail author
  • Rafał Fudała
  • Eva Holmström
  • Anders Sjöholm
  • Andrej Weintraub
  • Bożena Futoma-Kołoch
  • Gabriela Bugla-Płoskońska
  • Włodzimierz Doroszkiewicz
Open Access
Original Article

Abstract

Introduction

Proteus mirabilis bacilli play an important role in human urinary tract infections, bacteremia, and rheumatoid arthritis. The authors previously studied human complement C3 conversion by smooth-form P. mirabilis O10, O23, O30, and O43 lipopolysaccharides (LPSs) and showed that smooth Proteus LPSs fragmented C3 in a dose- and time-dependent manner. In the present study, one smooth P. mirabilis S1959 and its two polysaccharide-truncated LPSs isolated from an R mutant strain were used to study the C3 conversion.

Materials and Methods

The conversion of C3 to C3c by smooth and rough P. mirabilis LPSs was studied by capture ELISA and crossed immunoelectrophoresis. Proteins isolated from the outer membrane were analyzed by discontinuous sodium dodecyl sulfate gel electrophoresis.

Results

The smooth P. mirabilis S1959 (O3) strain was resistant to the bactericidal activity of human serum, in contrast to the Ra and Re mutant strains. The presence of an exposed core oligosaccharide in R110 LPS was not sufficient to protect the strain from serum-dependent killing. In addition to LPS structure, the outer-membrane proteins may also play roles in protecting the smooth P. mirabilis S1959 (O3) strain from the bactericidal action of serum. It was shown that the Ra P. mirabilis R110 and the Re P. mirabilis R45 mutants possess very different OMP compositions from that of the P. mirabilis S 1959 strain.

Conclusion

Regardless of the complement resistance of the P. mirabilis strains, the S1959, R110, and R45 LPSs fragmented C3 and induced C3c neo-antigen exposure. The use of complement-deficient human serum allows the conclusion that the Re-type P. mirabilis R45 LPS fragmented C3 by the antibody-independent classical pathway.

Keywords

Proteus mirabilis complement lipopolysaccharide outer-membrane protein 

References

  1. Alberti S, Marques G, Camprubi S et al (1993) C1q binding and activation of the complement classical pathway by Klebsiella pneumoniae outer membrane proteins. Infect Immun 61: 852–860PubMedGoogle Scholar
  2. Amano K, Cedzyński M, Swierzko AS et al (1996) Comparison of serological reactions of Rickettsiae-infected patients and rabbit anti-Proteus OX antibodies with Proteus OX2, OX19 and OXK lipopolysaccharides. Arch Immunol Ther Exp 44: 235–240Google Scholar
  3. Andersson J, Ekdahl KN, Lambris JD et al (2005) Binding of C3 fragments on top of adsorbed plasma proteins during complement activation on a model biomaterial surface. Biomaterial 26: 1477–1485CrossRefGoogle Scholar
  4. Becherer JD, Alsenz J, Lambris JD (1990) Molecular aspects of C3 interactions and structural functional analysis of C3 from different species. Curr Top Microbiol Immunol 153: 45–72PubMedGoogle Scholar
  5. Besucher HU, Brade V (1986) Lipopolysaccharides as complement inhibitors by complex formation with purified third complement component (C3). Immunobiology 173: 41–55Google Scholar
  6. Biedzka-Sarek M, Venho R, Skurnik M (2005) Role of YadA, Ail, and lipopolysaccharide in serum resistance of Yersinia enterocolitica serotype O:3. Infect Immun 73: 2232–2244PubMedCrossRefGoogle Scholar
  7. Blatteis CM, Li S, Li Z et al (2004) Complement is required for induction of endotoxic fever in guinea pigs and mice. J Therm Biol 29: 369–381CrossRefGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254PubMedCrossRefGoogle Scholar
  9. Burns SM, Hull SI (1998) Comparison of loss serum resistance by defined lipopolysaccharide mutants and an acapsular mutant of uropathogenic Escherichia coli O75:K5. Infect Immun 66: 4244–4253PubMedGoogle Scholar
  10. Chonn A, Cullis PR, Devine DV (1991) The role of surface changes in the activation of the classical and alternative pathways of complement by liposomes. J Immunol 146: 4234–4241PubMedGoogle Scholar
  11. Cisowska AG, Bugla-Ploskonska A, Gamian W et al (2005) Relationship between susceptibility to bactericidal action of serum and outer membrane protein patterns in E. coli K1 strains. Pol J Environ Stud 14: 476–482Google Scholar
  12. Doroszkiewicz W, Lachowicz TM (1989) Mechanism of antigenic variation in Shigella flexnerii bacilli. I. Selective lethal effect of normal sera on mixed population of S. flexnerii 1b serotype and its antigenic 3b mutant. Arch Immunol Ther Exp 37: 693–701Google Scholar
  13. Fischer MB, Prodeus AP, Nicholson-Weller A et al (1997) Increased susceptibility to endotoxin shock in complement C3-C4-deficient mice is corrected by C1 inhibitor replacement. J Immunol 159: 976–982PubMedGoogle Scholar
  14. Nordin Fredrikson G, Truedsson L, Sjöholm AG et al (1999) DNA analysis in MHC heterozygous patient with complete C4 deficiency – homozygosity for C4 gene deletion and C4 pseudogene. Exp Clin Immunogenet 8: 29–37Google Scholar
  15. Futoma-Kołoch B, Bugla-Płoskońska G, Doroszkiewicz W et al (2006) Survival of Proteus mirabilis O3 (S1959), O9 and O18 strains in normal human serum (NHS) correlates with the diversity of their outer membrane proteins (OMPs). Pol J Microbiol 55: 153–156PubMedGoogle Scholar
  16. Galanos C, Luderitz O, Westphal O (1969) A new method for the extraction of R-lipopolysaccharides. Eur J Biochem 9: 245–249PubMedCrossRefGoogle Scholar
  17. Gardiner JS, Keil LB, DeBari VA (1991) In vitro formation of complement activation products by lipopolysacharide chemotypes of Salmonella minnesota. Int Arch Allergy Appl Immunol 96: 51–54PubMedCrossRefGoogle Scholar
  18. Garred P, Mollnes TE, Lea T (1988) Quantification in enzyme-linked immunosorbent assay of a C3 neoepitope expressed on activated human complement factor C3. Scand J Immunol 27: 329–335PubMedCrossRefGoogle Scholar
  19. Holmskov-Nielsen JC, Jensenius B, Teisner B et al (1986) Measurements of C3 conversion by ELISA estimation of neo-determinants on the C3d moiety. J Immunol Methods 94: 1–6PubMedCrossRefGoogle Scholar
  20. Hostetter MK (1993) The third component of complement: new functions for an old friend. J Lab Clin Med 122: 491–496PubMedGoogle Scholar
  21. Hostetter MK, Gordon DL (1987) Biochemistry of C3 and related thioester proteins in infection and inflammation. Rev Infect Dis 9: 97–109PubMedGoogle Scholar
  22. Johnson U, Holmström E (1982) C3 fragmentation in human serum: formation of a mixed disulfide between C3d and albumin. Acta Pathol Microbiol Immunol Scand C 90: 321–326PubMedGoogle Scholar
  23. Joiner KA, Grossman N, Schmetz M et al (1986) C3 binds preferentially to long-chain lipopolysaccharide during alternative pathway activation by Salmonella monevideo. J Immunol 136: 710–715PubMedGoogle Scholar
  24. Kaca W, Literacka E, Sjoholm AG et al (2000) Complement activation by Proteus mirabilis negatively charged lipopolysaccharides. J Endotoxin Res 6: 223–234PubMedGoogle Scholar
  25. Kaca W, Roth R (1995) Activation of complement by human hemoglobin and by mixture of hemoglobin and bacterial endotoxin. Biochim Biophys Acta 1245: 49–56PubMedGoogle Scholar
  26. Klink M, Brzychcy M, Ziółkowski A et al (1998) The comparison of some biological activities of lipopolysaccharides obtained from smooth and rough Proteus mirabilis strains. Acta Microbiol Pol 47: 141–151PubMedGoogle Scholar
  27. Lachowicz TM, Doroszkiewicz W (1996) Sensitivity to complement as a factor of antigenic variation I Shigella flexnerii. Bull Pol Acad Biol Sci 44: 255–260Google Scholar
  28. Lachowicz TM, Doroszkiewicz W, Niedbach J (1999) Environment implication of lipopolysaccharide dependent normal serum sensitivity of Shigella flexnerii serotypes. Nova Acta Leopoldina 312: 235–243Google Scholar
  29. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685PubMedCrossRefGoogle Scholar
  30. Merino S, Nogueras MM, Aquilar A et al (1998) Activation the complement classical pathway (C1q binding) by mesophilic Aeromonas hydrophila outer membrane protein. Infect Immun 66: 3825–3831PubMedGoogle Scholar
  31. Mielnik G, Gamian A, Doroszkiewicz W (2001) Bactericidal activity of normal cord serum (NCS) against Gram-negative rods with sialic acid-containing lipopolysaccharides (LPS). FEMS Immunol Med Microbiol 31: 169–173PubMedCrossRefGoogle Scholar
  32. Munstermann M, Wiese A, Brabdenburg K et al (1999) Complement activation by bacterial surface glycolipids: a study with planar bilayer membranes. J Membr Biol 167: 223–232PubMedCrossRefGoogle Scholar
  33. Murphy TF, Bartos LC (1989) Surface-exposed and antigenically conserved determinants of outer membrane proteins of Branhamella catarrhalis. Infect Immun 57: 2938–2941PubMedGoogle Scholar
  34. Nawrot U, Mokracka-Latajka G, Grzybek-Hryncewicz J et al (1995) Bactericidal activity of normal human serum against Morganella, Proteus and Providencia strains. Acta Microbiol Pol 44: 55–61PubMedGoogle Scholar
  35. Ohta M, Okada M, Yamashina I et al (1990) The mechanism of carbohydrate-mediated complement activation by the serum mannan-binding protein. J Biol Chem 265: 1980–1984PubMedGoogle Scholar
  36. Pilz D, Vocke T, Heesemann J et al (1992) Mechanism of YadA-mediated serum resistance of Yersinia enterocolitica serotype O3. Infect Immun 60: 189–195PubMedGoogle Scholar
  37. Prasadarao NV, Blom AM, Villoutreix BO et al (2002) A novel interaction of outer membrane protein A with C4b binding protein mediates serum resistance of Escherichia coli K1. J Immunol 169: 6352–6360PubMedGoogle Scholar
  38. Ram S, Sharma AK, Simpson SD et al (1998) A novel sialyc acid binding site on factor H mediates serum resistance of silalylated Neisseria gonorrhorea. J Exp Med 187: 743–752PubMedCrossRefGoogle Scholar
  39. Reeves P (1995) Role of O-antigen variation on the immune response. Trends Microbiol 3: 381–386PubMedCrossRefGoogle Scholar
  40. Różalski A, Sidorczyk Z, Kotelko K (1997) Potential virulence factors of Proteus bacilli. Microbiol Mol Biol Rev 61: 65–89PubMedGoogle Scholar
  41. Sjöholm AG, Braconier JH, Söderstrström C (1982) Properdin deficiency in a family with fulminant meningococcal infections. Clin Exp Immunol 50: 291–297PubMedGoogle Scholar
  42. Taylor PW (1992) Complement-mediated killing of susceptible Gram-negative bacteria: an elusive mechanism. Exp Clin Immunogenet 9: 48–56PubMedGoogle Scholar
  43. Vinogradov E, Radziejewska-Lebrecht J, Kaca W (2000) The structure of the carbohydrate backbone of core-lipid A region of the lipopolysaccharide from Proteus mirabilis wild-type strain S1959 (serotype O3) and its Ra mutant R110/1959. Eur J Biochem 267: 262–269PubMedCrossRefGoogle Scholar
  44. Vinogradov EV, Thomas-Oates JE, Brade H et al (1994) Structural investigation of the lipopolysaccharide from Proteus mirabilis R45 (Re-chemotype). J Endotoxin Res 1: 199–206Google Scholar
  45. Vukajlovich SW, Hoffman J, Morrison DC (1987) Activation of human serum complement by bacterial lipopolysaccharides: structural requirements for antibody independent activation of classical and alternative pathways. Mol Immunol 24: 319–331PubMedCrossRefGoogle Scholar
  46. Westphal O, Jann K (1965) Bacterial lipopolysaccharides: extraction with phenol-water and further application of the procedure. Methods Carbohydr Chem 5: 83–89Google Scholar
  47. Ziółkowski A, Shashkov AS, Swierzko AS et al (1997) Structures of the O-antigen of Proteus bacilli belonging to OX group (serogroup O1-O3) used in Weil-Felix test. FEBS Lett 411: 221–224PubMedCrossRefGoogle Scholar

Copyright information

© L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2009

Authors and Affiliations

  • Wiesław Kaca
    • 1
    • 2
  • Michał Arabski
    • 2
    Email author
  • Rafał Fudała
    • 2
  • Eva Holmström
    • 3
  • Anders Sjöholm
    • 3
  • Andrej Weintraub
    • 1
  • Bożena Futoma-Kołoch
    • 4
  • Gabriela Bugla-Płoskońska
    • 4
  • Włodzimierz Doroszkiewicz
    • 4
  1. 1.Division of Clinical BacteriologyHuddinge University Hospital, Karolinska InstituteHuddingeSweden
  2. 2.Department of Microbiology, Institute of BiologyJan Kochanowski UniversityKielcePoland
  3. 3.Division of Microbiology, Immunology, and GlycobiologyLund UniversityLundSweden
  4. 4.Institute of Genetics and MicrobiologyWrocław UniversityWrocławPoland

Personalised recommendations