Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1979 Dec;26(3):808–814. doi: 10.1128/iai.26.3.808-814.1979

Quantitation of the Third Component of Human Complement Attached to the Surface of Opsonized Bacteria: Opsonin-Deficient Sera and Phagocytosis-Resistant Strains

Henri A Verbrugh 1, Willemien C van Dijk 1, Marijke E van Erne 1, Roel Peters 1, Phillip K Peterson 2, Jan Verhoef 1
PMCID: PMC414690  PMID: 393633

Abstract

The role of the third component of human complement (C3) in the opsonization of bacteria in nonimmune human sera was evaluated. The amount of C3 that becomes attached to the surface of bacteria upon incubation in serum was measured in a quantitative fluorescent immunoassay using fluorescein-conjugated monospecific antiserum to human C3. The intensity of the fluorescence from opsonized bacteria was found to be directly proportional to the absolute amount of C3 fixed, and this enabled the detection of as few as 300 molecules of bound C3 per bacterium. In normal serum the rate of C3 fixation was closely correlated with an increase in opsonization of the bacteria for human PMNs. Both C3 fixation and opsonization were maximal after 15 min of incubation. C3 fixation was also observed, albeit at a significantly slower rate, in human serum with a nonfunctional classical pathway but an intact alternative complement pathway and in serum deficient in immunoglobulins. Again, the kinetics of C3 fixation correlated with bacterial opsonization. Using a total of 21 strains of several bacterial species, including Staphylococcus aureus and Escherichia coli, encapsulation of bacteria was found to interfere with the process of C3 fixation in normal human serum, rendering these organisms resistant to subsequent phagocytosis by human polymorphonuclear leukocytes.

Full text

PDF
808

Selected References

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

  1. Alexander J. W., McClellan M. A., Ogle C. K., Ogle J. D. Consumptive opsoninopathy: possible pathogenesis in lethal and opportunistic infections. Ann Surg. 1976 Dec;184(6):672–678. doi: 10.1097/00000658-197612000-00002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alper C. A., Abramson N., Johnston R. B., Jr, Jandl J. H., Rosen F. S. Increased susceptibility to infection associated with abnormalities of complement-mediated functions and of the third component of complement (C3). N Engl J Med. 1970 Feb 12;282(7):350–354. doi: 10.1056/nejm197002122820701. [DOI] [PubMed] [Google Scholar]
  3. Biggar W. D., Holmes B., Good R. A. Opsonic defect in patients with cystic fibrosis of the pancreas. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1716–1719. doi: 10.1073/pnas.68.8.1716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark R. A., Klebanoff S. J. Role of the classical and alternative complement pathways in chemotaxis and opsonization: studies of human serum deficient in C4. J Immunol. 1978 Apr;120(4):1102–1108. [PubMed] [Google Scholar]
  6. Dalmasso A. P., Müller-Eberhard H. J. Physico-chemical characteristics of the third and fourth component of complement after dissociation from complement-cell complexes. Immunology. 1967 Sep;13(3):293–305. [PMC free article] [PubMed] [Google Scholar]
  7. Forsgren A., Quie P. G. Influence of the alternate complement pathway in opsonization of several bacterial species. Infect Immun. 1974 Aug;10(2):402–404. doi: 10.1128/iai.10.2.402-404.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giebink G. S., Verhoef J., Peterson P. K., Quie P. G. Opsonic requirements for phagocytosis of Streptococcus pneumoniae types VI, XVIII, XXIII, and XXV. Infect Immun. 1977 Nov;18(2):291–297. doi: 10.1128/iai.18.2.291-297.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gigli I., Nelson R. A., Jr Complement dependent immune phagocytosis. I. Requirements for C'1, C'4, C'2, C'3. Exp Cell Res. 1968 Jul;51(1):45–67. doi: 10.1016/0014-4827(68)90158-4. [DOI] [PubMed] [Google Scholar]
  10. Gillis T. P., Thompson J. J. Quantitative fluorescent immunoassay of antibodies to, and surface antigens of, Actinomyces viscosus. J Clin Microbiol. 1978 Feb;7(2):202–208. doi: 10.1128/jcm.7.2.202-208.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hand W. L., King N. L. Serum opsonization of salmonella in sickle cell anemia. Am J Med. 1978 Mar;64(3):388–395. doi: 10.1016/0002-9343(78)90217-6. [DOI] [PubMed] [Google Scholar]
  12. Howard C. J., Glynn A. A. The virulence for mice of strains of Escherichia coli related to the effects of K antigens on their resistance to phagocytosis and killing by complement. Immunology. 1971 May;20(5):767–777. [PMC free article] [PubMed] [Google Scholar]
  13. Jasin H. E. Human heat labile opsonins: evidence for their mediation via the alternate pathway of complement activation. J Immunol. 1972 Jul;109(1):26–31. [PubMed] [Google Scholar]
  14. Johnson F. R., Agnello V., Williams R. C., Jr Opsonic activity in human serum deficient in C2. J Immunol. 1972 Jul;109(1):141–145. [PubMed] [Google Scholar]
  15. Johnston R. B., Jr, Klemperer M. R., Alper C. A., Rosen F. S. The enhancement of bacterial phagocytosis by serum. The role of complement components and two cofactors. J Exp Med. 1969 Jun 1;129(6):1275–1290. doi: 10.1084/jem.129.6.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Melly M. A., Duke L. J., Liau D. F., Hash J. H. Biological properties of the encapsulated Staphylococcus aureus M. Infect Immun. 1974 Aug;10(2):389–397. doi: 10.1128/iai.10.2.389-397.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mickenberg I. D., Root R. K., Wolff S. M. Leukocytic function in hypogammaglobulinemia. J Clin Invest. 1970 Aug;49(8):1528–1538. doi: 10.1172/JCI106370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Peterson P. K., Wilkinson B. J., Kim Y., Schmeling D., Quie P. G. Influence of encapsulation on staphylococcal opsonization and phagocytosis by human polymorphonuclear leukocytes. Infect Immun. 1978 Mar;19(3):943–949. doi: 10.1128/iai.19.3.943-949.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pondman K. W., Hannema A., Cormane R., Stoop J. W. Ongewone complementdeficiëntie bij een patiëntje met lupus erythematosus disseminatus. Ned Tijdschr Geneeskd. 1969 Aug 23;113(34):1462–1469. [PubMed] [Google Scholar]
  20. Schreiber R. D., Pangburn M. K., Lesavre P. H., Müller-Eberhard H. J. Initiation of the alternative pathway of complement: recognition of activators by bound C3b and assembly of the entire pathway from six isolated proteins. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3948–3952. doi: 10.1073/pnas.75.8.3948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shin H. S., Smith M. R., Wood W. B., Jr Heat labile opsonins to pneumococcus. II. Involvement of C3 and C5. J Exp Med. 1969 Dec 1;130(6):1229–1241. doi: 10.1084/jem.130.6.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith H. Microbial surfaces in relation to pathogenicity. Bacteriol Rev. 1977 Jun;41(2):475–500. doi: 10.1128/br.41.2.475-500.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Smith M. R., Shin H. S., Wood W. B., Jr Natural immunity to bacterial infections: the relation of complement to heat-labile opsonins. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1151–1156. doi: 10.1073/pnas.63.4.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stevens P., Huang S. N., Welch W. D., Young L. S. Restricted complement activation by Escherichia coli with the K-1 capsular serotype: a possible role in pathogenicity. J Immunol. 1978 Dec;121(6):2174–2180. [PubMed] [Google Scholar]
  25. Stossel T. P., Field R. J., Gitlin J. D., Alper C. A., Rosen F. S. The opsonic fragment of the third component of human complement (C3). J Exp Med. 1975 Jun 1;141(6):1329–1347. doi: 10.1084/jem.141.6.1329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stossel T. P. Phagocytosis: recognition and ingestion. Semin Hematol. 1975 Jan;12(1):83–116. [PubMed] [Google Scholar]
  27. Van Dijk W. C., Verbrugh H. A., Peters R., Van Der Tol M. E., Peterson P. K., Verhoef J. Escherichia coli K antigen in relation to serum-induced lysis and phagocytosis. J Med Microbiol. 1979 Feb;12(1):123–130. doi: 10.1099/00222615-12-1-123. [DOI] [PubMed] [Google Scholar]
  28. Verbrugh H. A., Van Dijk W. C., Peters R., Van Der Tol M. E., Peterson P. K., Verhoef J. Staphylococcus aureus opsonization mediated via the classical and alternative complement pathways. A kinetic study using MgEGTA chelated serum and human sera deficient in IgG and complement factors C1s and C2. Immunology. 1979 Mar;36(3):391–397. [PMC free article] [PubMed] [Google Scholar]
  29. Verhoef J., Peterson P., Kim Y., Sabath L. D., Quie P. G. Opsonic requirements for staphylococcal phagocytosis. Heterogeneity among strains. Immunology. 1977 Aug;33(2):191–197. [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES