Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Jun;40(3):967–976. doi: 10.1128/iai.40.3.967-976.1983

Inhibition of neutrophil function by fluid phase C3b of complement.

J D Ogle, C K Ogle, J W Alexander
PMCID: PMC348146  PMID: 6221999

Abstract

A high-molecular-weight fragment of C3 was isolated from normal human serum by column chromatography, was generated by incubation of serum at 37 degrees C with inulin, and was produced from highly purified C3 by limited digestion with trypsin. This product was shown to inhibit the antibacterial function of neutrophils by using Escherichia coli O75 as the main test organism. The inhibitor reacted with anti-C3b and anti-C3c, but not with anti-C3B (anti-native C3) or anti-C3a. The manner of preparation of the inhibitor, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern, and the amino acid composition of the inhibitor indicated that it was fluid phase C3b. The inhibitor of neutrophil function (fluid phase C3b) was shown to bind to C3b receptors or acceptors on sheep erythrocytes in a model system.

Full text

PDF
967

Images in this article

969Image
on p.969

Selected References

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

  1. Alexander J. W., Meakins J. L. A physiological basis for the development of opportunistic infections in man. Ann Surg. 1972 Sep;176(3):273–287. doi: 10.1097/00000658-197209000-00003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alexander J. W., Windhorst D. B., Good R. A. Improved tests for the evaluation of neutrophil function in human disease. J Lab Clin Med. 1968 Jul;72(1):136–148. [PubMed] [Google Scholar]
  3. Arnaout M. A., Melamed J., Tack B. F., Colten H. R. Characterization of the human complement (c3b) receptor with a fluid phase C3b dimer. J Immunol. 1981 Oct;127(4):1348–1354. [PubMed] [Google Scholar]
  4. Berger M., Gaither T. A., Hammer C. H., Frank M. M. Lack of binding of human C3, in its native state, to C3b receptors. J Immunol. 1981 Oct;127(4):1329–1334. [PubMed] [Google Scholar]
  5. Budzko D. B., Bokisch V. A., Müller-Eberhard H. J. A fragment of the third component of human complement with anaphylatoxin activity. Biochemistry. 1971 Mar 30;10(7):1166–1172. doi: 10.1021/bi00783a011. [DOI] [PubMed] [Google Scholar]
  6. Chaplin H., Jr, Hoffmann N. L., Monroe M. C. A simple method for preparation of C3c fragment from trypsinized serum-reacted zymosan. Transfusion. 1980 May-Jun;20(3):341–347. doi: 10.1046/j.1537-2995.1980.20380214904.x. [DOI] [PubMed] [Google Scholar]
  7. Czop J. K., Austen K. F. Functional discrimination by human monocytes between their C3b receptors and their recognition units for particulate activators of the alternative complement pathway. J Immunol. 1980 Jul;125(1):124–128. [PubMed] [Google Scholar]
  8. Dierich M. P., Bokisch V. A. Receptor-binding sites on C3 and C3b. J Immunol. 1977 Jun;118(6):2145–2150. [PubMed] [Google Scholar]
  9. Dierich M. P., Landen B. Complement bridges between cells analysis of a possible cell-cell interaction mechanism. J Exp Med. 1977 Dec 1;146(6):1484–1499. doi: 10.1084/jem.146.6.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. ERLANGER B. F., KOKOWSKY N., COHEN W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys. 1961 Nov;95:271–278. doi: 10.1016/0003-9861(61)90145-x. [DOI] [PubMed] [Google Scholar]
  11. Ehlenberger A. G., Nussenzweig V. The role of membrane receptors for C3b and C3d in phagocytosis. J Exp Med. 1977 Feb 1;145(2):357–371. doi: 10.1084/jem.145.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  13. Fearon D. T. Identification of the membrane glycoprotein that is the C3b receptor of the human erythrocyte, polymorphonuclear leukocyte, B lymphocyte, and monocyte. J Exp Med. 1980 Jul 1;152(1):20–30. doi: 10.1084/jem.152.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fontaine M., Joisel F., Dumouchel L. Preparation of an R3 reagent (serum depleted of the third component of human complement (C3)) by immunoadsorption. Application to the hemolytic assay of human C3. J Immunol Methods. 1980;33(2):145–158. doi: 10.1016/s0022-1759(80)80005-6. [DOI] [PubMed] [Google Scholar]
  15. Ghebrehiwet B., Müller-Eberhard H. J. C3e: an acidic fragment of human C3 with leukocytosis-inducing activity. J Immunol. 1979 Aug;123(2):616–621. [PubMed] [Google Scholar]
  16. Gitlin J. D., Rosen F. S., Lachmann P. J. The mechanism of action of the C3b inactivator (conglutinogen-activating factor) on its naturally occurring substrate, the major fragment of the third component of complement (C3b). J Exp Med. 1975 May 1;141(5):1221–1226. doi: 10.1084/jem.141.5.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goldstein I. M., Brai M., Osler A. G., Weissmann G. Lysosomal enzyme release from human leukocytes: mediation by the alternate pathway of complement activation. J Immunol. 1973 Jul;111(1):33–37. [PubMed] [Google Scholar]
  18. Hammer C. H., Wirtz G. H., Renfer L., Gresham H. D., Tack B. F. Large scale isolation of functionally active components of the human complement system. J Biol Chem. 1981 Apr 25;256(8):3995–4006. [PubMed] [Google Scholar]
  19. Hugli T. E. Human anaphylatoxin (C3a) from the third component of complement. Primary structure. J Biol Chem. 1975 Nov 10;250(21):8293–8301. [PubMed] [Google Scholar]
  20. Isenman D. E., Kells D. I., Cooper N. R., Müller-Eberhard H. J., Pangburn M. K. Nucleophilic modification of human complement protein C3: correlation of conformational changes with acquisition of C3b-like functional properties. Biochemistry. 1981 Jul 21;20(15):4458–4467. doi: 10.1021/bi00518a034. [DOI] [PubMed] [Google Scholar]
  21. Janatova J., Lorenz P. E., Schechter A. N., Prahl J. W., Tack B. F. Third component of human complement: appearance of a sulfhydryl group following chemical or enzymatic inactivation. Biochemistry. 1980 Sep 16;19(19):4471–4478. [PubMed] [Google Scholar]
  22. Kunkel S. L., Kreutzer D. L., Goralnick S., Ward P. A. Purification of the third and fifth components of human complement: application of hydrophobic chromatography. J Immunol Methods. 1980;35(3-4):337–351. doi: 10.1016/0022-1759(80)90259-8. [DOI] [PubMed] [Google Scholar]
  23. Lambris J. D., Ross G. D. Assay of membrane complement receptors (CR1 and CR2) with C3b- and C3d-coated fluorescent microspheres. J Immunol. 1982 Jan;128(1):186–189. [PubMed] [Google Scholar]
  24. Law S. K., Levine R. P. Interaction between the third complement protein and cell surface macromolecules. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2701–2705. doi: 10.1073/pnas.74.7.2701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Leijh P. C., van den Barselaar M. T., Daha M. R., van Furth R. Participation of immunoglobulins and complement components in the intracellular killing of Staphylococcus aureus and Escherichia coli by human granulocytes. Infect Immun. 1981 Sep;33(3):714–724. doi: 10.1128/iai.33.3.714-724.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  27. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  28. Melamed J., Arnaout M. A., Colten H. R. Complement (C3b) interaction with the human granulocyte receptor: correlation of binding of fluid-phase radiolabeled ligand with histaminase release. J Immunol. 1982 May;128(5):2313–2318. [PubMed] [Google Scholar]
  29. Molenaar J. L., ten Velde A., Pondman K. W. Anti-C3a, anti-C3B (anti-B) and their reaction with the anaphylatoxic fragment C3a. J Immunol. 1973 Mar;110(3):702–708. [PubMed] [Google Scholar]
  30. Müller-Eberhard H. J. Complement. Annu Rev Biochem. 1975;44:697–724. doi: 10.1146/annurev.bi.44.070175.003405. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Newman S. L., Johnston R. B., Jr Role of binding through C3b and IgG in polymorphonuclear neutrophil function: studies with trypsin-generated C3b. J Immunol. 1979 Oct;123(4):1839–1846. [PubMed] [Google Scholar]
  33. Nicholson A., Brade V., Lee G. D., Shin H. S., Mayer M. M. Kinetic studies of the formation of the properdin system enzymes on zymosan: evidence that nascent C3b controls the rate of assembly. J Immunol. 1974 Mar;112(3):1115–1123. [PubMed] [Google Scholar]
  34. Pangburn M. K., Müller-Eberhard H. J. Relation of putative thioester bond in C3 to activation of the alternative pathway and the binding of C3b to biological targets of complement. J Exp Med. 1980 Oct 1;152(4):1102–1114. doi: 10.1084/jem.152.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pangburn M. K., Schreiber R. D., Müller-Eberhard H. J. Human complement C3b inactivator: isolation, characterization, and demonstration of an absolute requirement for the serum protein beta1H for cleavage of C3b and C4b in solution. J Exp Med. 1977 Jul 1;146(1):257–270. doi: 10.1084/jem.146.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ross G. D. Analysis of the different types of leukocyte membrane complement receptors and their interaction with the complement system. J Immunol Methods. 1980;37(3-4):197–211. doi: 10.1016/0022-1759(80)90307-5. [DOI] [PubMed] [Google Scholar]
  37. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  38. Schmitt M., Mussel H. H., Dierich M. P. Qualitative and quantitative assessment of C3-receptor reactivities on lymphoid and phagocytic cells. J Immunol. 1981 May;126(5):2042–2047. [PubMed] [Google Scholar]
  39. Tack B. F., Harrison R. A., Janatova J., Thomas M. L., Prahl J. W. Evidence for presence of an internal thiolester bond in third component of human complement. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5764–5768. doi: 10.1073/pnas.77.10.5764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tack B. F., Morris S. C., Prahl J. W. Third component of human complement: structural analysis of the polypeptide chains of C3 and C3b. Biochemistry. 1979 Apr 17;18(8):1497–1503. doi: 10.1021/bi00575a017. [DOI] [PubMed] [Google Scholar]
  41. Verbrugh H. A., van Dijk W. C., van Erne M. E., Peters R., Peterson P. K., Verhoef J. Quantitation of the third component of human complement attached to the surface of opsonized bacteria: opsonin-deficient sera and phagocytosis-resistant strains. Infect Immun. 1979 Dec;26(3):808–814. doi: 10.1128/iai.26.3.808-814.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  43. Wong L., Wilson J. D. The identification of Fc and C3 receptors on human neutrophils. J Immunol Methods. 1975 Apr;7(1):69–76. doi: 10.1016/0022-1759(75)90131-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES