Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1968 Feb;95(2):397–405. doi: 10.1128/jb.95.2.397-405.1968

Interactions of the Complement System with Native and Chemically Modified Endotoxins

H Gewurz 1,2, S E Mergenhagen 1,2, A Nowotny 1,2, J K Phillips 1,2
PMCID: PMC252032  PMID: 4170751

Abstract

Endotoxic lipopolysaccharides (LPS) isolated from Serratia marcescens, Veillonella alcalescens, and Salmonella typhosa were potent in their ability to induce fixation of complement (C′) in normal guinea pig, rabbit, mouse, and human serum. The C′-fixing ability of LPS was pronounced even when assays were performed in undiluted serum, and was lost after each of four chemical modifications which resulted in loss of biological toxicities. The detoxification procedures had in common the cleavage of ester-bound, long-chain carboxylic acids. The ability of biologically active LPS to fix C′ in normal guinea pig serum was reflected chiefly in dramatic uptake of classical C′3 (C′3t); fixation of C′1, C′4, and C′2 was virtually undetectable. Hence, it was the capacity for fixation of C′3t which was lost most overtly during detoxification. Addition of immune serum to the assay mixtures resulted in detectable fixation of C′1 and C′4. Biologically active LPS also fixed more of these components than did detoxified LPS. Immune serum restored the ability of detoxified LPS to fix C′3t, but whether this is by the original pathway is not yet clear. We concluded that the loss of certain biological activities and the loss of ability to fix C′3t in normal serum after LPS detoxification involved loss or rearrangement of substrates on LPS which either initiated or supported, or both, its interaction with the complement system. It was apparent that the ability to fix C′ can serve as a valuable in vitro indicator of the integrity of the toxic conformation of biologically active LPS membrane fragments. These experiments supported the hypothesis that certain of the biological activities induced by endotoxins are mediated via the complement system.

Full text

PDF
397

Selected References

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

  1. BLADEN H. A., MERGENHAGEN S. E. ULTRASTRUCTURE OF VEILLONELLA AND MORPHOLOGICAL CORRELATION OF AN OUTER MEMBRANE WITH PARTICLES ASSOCIATED WITH ENDOTOXIC ACTIVITY. J Bacteriol. 1964 Nov;88:1482–1492. doi: 10.1128/jb.88.5.1482-1492.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLOCH K. J., KOURILSKY F. M., OVARY Z., BENACERRAF B. Properties of guinea pig 7S antibodies. III. Identification of antibodies involved in complement fixation and hemolysis. J Exp Med. 1963 Jun 1;117:965–981. doi: 10.1084/jem.117.6.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bladen H. A., Gewurz H., Mergenhagen S. E. Interactions of the complement system with the surface and endotoxic lipopolysaccharide of Veillonella alcalescens. J Exp Med. 1967 May 1;125(5):767–786. doi: 10.1084/jem.125.5.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cremer N. E., Riggs J. L., Lennette E. H., Jensen F. W. Nonspecific Binding of Complement by Digestion Fragments from Antiviral Gamma Globulin. Science. 1965 Jul 2;149(3679):84–85. doi: 10.1126/science.149.3679.84. [DOI] [PubMed] [Google Scholar]
  5. De Petris S. Ultrastructure of the cell wall of Escherichia coli and chemical nature of its constituent layers. J Ultrastruct Res. 1967 Jul;19(1):45–83. doi: 10.1016/s0022-5320(67)80059-5. [DOI] [PubMed] [Google Scholar]
  6. Dias Da Silva W., Lepow I. H. Complement as a mediator of inflammation. II. Biological properties of anaphylatoxin prepared with purified components of human complement. J Exp Med. 1967 May 1;125(5):921–946. doi: 10.1084/jem.125.5.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Forsgren A., Sjöquist J. "Protein A" from Staphylococcus aureus. 3. Reaction with rabbit gamma-globulin. J Immunol. 1967 Jul;99(1):19–24. [PubMed] [Google Scholar]
  8. Freedman H. H., Sultzer B. M. Dissociation of antitoxic and immunogenic activities of endotoxin by acetylation. Ann N Y Acad Sci. 1966 Jun 30;133(2):580–585. doi: 10.1111/j.1749-6632.1966.tb52390.x. [DOI] [PubMed] [Google Scholar]
  9. GILBERT V. E., BRAUDE A. I. Reduction of serum complement in rabbits after injection of endotoxin. J Exp Med. 1962 Oct 1;116:477–490. doi: 10.1084/jem.116.4.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gewurz H., Page A. R., Pickering R. J., Good R. A. Complement activity and inflammatory neutrophil exudation in man. Studies in patients with glomerulonephritis, essential hypocomplementemia and agammaglobulinemia. Int Arch Allergy Appl Immunol. 1967;32(1):64–90. doi: 10.1159/000229917. [DOI] [PubMed] [Google Scholar]
  11. ISHIZAKA T., ISHIZAKA K. Biological activities of aggregated gamma globulin. I. Skin reactive and complement-fixing properties of heat denatured gamma globulin. Proc Soc Exp Biol Med. 1959 Aug-Sep;101:845–850. doi: 10.3181/00379727-101-25116. [DOI] [PubMed] [Google Scholar]
  12. Ishizaka T., Ishizaka K., Borsos T., Rapp H. C'-1 fixation by human isoagglutinins: fixation of C'-1 by gamma-G and gamma-M but not by gamma-A antibody. J Immunol. 1966 Nov;97(5):716–726. [PubMed] [Google Scholar]
  13. JOHNSON A. G., NOWOTNY A. RELATIONSHIP OF STRUCTURE TO FUNCTION IN BACTERIAL O ANTIGENS. III. BIOLOGICAL PROPERTIES OF ENDOTOXOIDS. J Bacteriol. 1964 Apr;87:809–814. doi: 10.1128/jb.87.4.809-814.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jensen J. Anaphylatoxin in its relation to the complement system. Science. 1967 Mar 3;155(3766):1122–1123. doi: 10.1126/science.155.3766.1122. [DOI] [PubMed] [Google Scholar]
  15. KOSTKA J., STERZL J. The action of endotoxin on complement. Folia Microbiol (Praha) 1962 May;7:191–192. doi: 10.1007/BF02928241. [DOI] [PubMed] [Google Scholar]
  16. LEPOW I. H., LEON M. A. Interaction of a serum inhibitor of C'I-esterase with intermediate complexes of the immune haemolytic system. I. Specificity of inhibition of C'I activity associated with intermediate complexes. Immunology. 1962 Mar;5:222–234. [PMC free article] [PubMed] [Google Scholar]
  17. MAYER M. M., LEVINE L. Kinetic studies on immune hemolysis. IV. Rate determination of the Mg++ and terminal reaction steps. J Immunol. 1954 Jun;72(6):516–530. [PubMed] [Google Scholar]
  18. MUELLER-EBERHARD H. J., LEPOW I. H. C'1 ESTERASE EFFECT ON ACTIVITY AND PHYSICOCHEMICAL PROPERTIES OF THE FOURTH COMPONENT OF COMPLEMENT. J Exp Med. 1965 May 1;121:819–833. doi: 10.1084/jem.121.5.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. MUSCHEL L. H., SCHMOKER K., WEBB P. M. ANTICOMPLEMENTARY ACTION OF ENDOTOXIN. Proc Soc Exp Biol Med. 1964 Dec;117:639–643. doi: 10.3181/00379727-117-29657. [DOI] [PubMed] [Google Scholar]
  20. Mergenhagen S. E., Bladen H. A., Hsu K. C. Electron microscopic localization of endotoxic lipopolysaccharide in gram-negative organisms. Ann N Y Acad Sci. 1966 Jun 30;133(2):279–291. doi: 10.1111/j.1749-6632.1966.tb52371.x. [DOI] [PubMed] [Google Scholar]
  21. Muller-Eberhard H. J., Nilsson U. R., Dalmasso A. P., Polley M. J., Calcott M. A. A molecular concept of immune cytolysis. Arch Pathol. 1966 Sep;82(3):205–217. [PubMed] [Google Scholar]
  22. Müllerèberhard H. J., Dalmasso A. P., Calcott M. A. The reaction mechanism of beta-1C-globulin (C'3) in immune hemolysis. J Exp Med. 1966 Jan 1;123(1):33–54. doi: 10.1084/jem.123.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. NETER E., WESTPHAL O., LUDERITZ O., GORZYNSKI E. A., EICHENBERGER E. Studies of enterobacterial lipopolysaccharides; effects of heat and chemicals on erythrocyte-modifying, antigenic, toxic and pyrogenic properties. J Immunol. 1956 May;76(5):377–385. [PubMed] [Google Scholar]
  24. NOLL H., BRAUDE A. I. Preparation and biological properties of a chemically modified Escherichia coli endotoxin of high immunogenic potency and low toxicity. J Clin Invest. 1961 Nov;40:1935–1951. doi: 10.1172/JCI104419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. NOWOTNY A. M., THOMAS S., DURON O. S., NOWOTNY A. Relation of structure to function in bacterial O antigens. I. Isolation methods. J Bacteriol. 1963 Feb;85:418–426. doi: 10.1128/jb.85.2.418-426.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nelson R. A., Jr, Jensen J., Gigli I., Tamura N. Methods for the separation, purification and measurement of nine components of hemolytic complement in guinea-pig serum. Immunochemistry. 1966 Mar;3(2):111–135. doi: 10.1016/0019-2791(66)90292-8. [DOI] [PubMed] [Google Scholar]
  27. OSLER A. G., RANDALL H. G., HILL B. M., OVARY Z. Studies on the mechanism of hypersensitivity phenomena. III. The participation of complement in the formation of anaphylatoxin. J Exp Med. 1959 Aug 1;110(2):311–339. doi: 10.1084/jem.110.2.311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. OSLER A. G., STRAUSS J. H., MAYER M. M. Diagnostic complement fixation. I. A method. Am J Syph Gonorrhea Vener Dis. 1952 Mar;36(2):140–153. [PubMed] [Google Scholar]
  29. PEARLMAN D. S., SAUERS J. B., TALMAGE D. W. THE EFFECT OF ADJUVANT AMOUNTS OF ENDOTOXINS ON THE SERUM HEMOLYTIC COMPLEMENT ACTIVITY IN RABBITS. J Immunol. 1963 Dec;91:748–756. [PubMed] [Google Scholar]
  30. PILLEMER L., SCHOENBERG M. D., BLUM L., WURZ L. Properdin system and immunity. II. Interaction of the properdin system with polysaccharides. Science. 1955 Sep 23;122(3169):545–549. doi: 10.1126/science.122.3169.545. [DOI] [PubMed] [Google Scholar]
  31. Rothfield L., Horne R. W. Reassociation of purified lipopolysaccharide and phospholipid of the bacterial cell envelope: electron microscopic and monolayer studies. J Bacteriol. 1967 May;93(5):1705–1721. doi: 10.1128/jb.93.5.1705-1721.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. SPINK W. W., DAVIS R. B., POTTER R., CHARTRAND S. THE INITIAL STAGE OF CANINE ENDOTOXIN SHOCK AS AN EXPRESSION OF ANAPHYLACTIC SHOCK: STUDIES ON COMPLEMENT TITERS AND PLASMA HISTAMINE CONCENTRATIONS. J Clin Invest. 1964 Apr;43:696–704. doi: 10.1172/JCI104954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. SPINK W. W. Endotoxin shock. Ann Intern Med. 1962 Oct;57:538–552. doi: 10.7326/0003-4819-57-4-538. [DOI] [PubMed] [Google Scholar]
  34. Shands J. W., Jr, Graham J. A., Nath K. The morphologic structure of isolated bacterial lipopolysaccharide. J Mol Biol. 1967 Apr 14;25(1):15–21. doi: 10.1016/0022-2836(67)90275-6. [DOI] [PubMed] [Google Scholar]
  35. Skarnes R. C. Non specific hemolysis of erythrocytes modified with bacterial endotoxins. Ann Inst Pasteur (Paris) 1965 Jul;109(1):66–79. [PubMed] [Google Scholar]
  36. TERRY W. D., BORSOS T., RAPP H. J. DIFFERENCES IN SERUM COMPLEMENT ACTIVITY AMONG INBRED STRAINS OF MICE. J Immunol. 1964 Apr;92:576–578. [PubMed] [Google Scholar]
  37. Treffers H. P. The Detoxification by Acetylation of Soluble Antigens From Shigella dysenteriae (Shiga) and E. typhosa. Science. 1946 Mar 29;103(2674):387–389. doi: 10.1126/science.103.2674.387. [DOI] [PubMed] [Google Scholar]
  38. Tripodi D., Nowotny A. Relation of structure to function in bacterial O-antigens. V. Nature of active sites in endotoxic lipopolysaccharides of Serratia marcescens. Ann N Y Acad Sci. 1966 Jun 30;133(2):604–621. doi: 10.1111/j.1749-6632.1966.tb52392.x. [DOI] [PubMed] [Google Scholar]
  39. WARD P. A., COCHRANE C. G., MUELLER-EBERHARD H. J. THE ROLE OF SERUM COMPLEMENT IN CHEMOTAXIS OF LEUKOCYTES IN VITRO. J Exp Med. 1965 Aug 1;122:327–346. doi: 10.1084/jem.122.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ward P. A., Cochrane C. G., Muller-Eberhard H. J. Further studies on the chemotactic factor of complement and its formation in vivo. Immunology. 1966 Aug;11(2):141–153. [PMC free article] [PubMed] [Google Scholar]
  41. ZWEIFACH B. W., JANOFF A. BACTERIAL ENDOTOXEMIA. Annu Rev Med. 1965;16:201–220. doi: 10.1146/annurev.me.16.020165.001221. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES