Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1982 Mar;35(3):929–936. doi: 10.1128/iai.35.3.929-936.1982

Characterization of Opsonins for Bacteroides fragilis in Immune Sera Collected from Experimentally Infected Mice

Thomas M Ellis 1,†, James T Barrett 1
PMCID: PMC351136  PMID: 7068222

Abstract

Serum collected from mice experimentally infected with Bacteroides fragilis 23745 (immune serum) was analyzed for its ability to opsonize the in vitro ingestion of this organism by mouse peritoneal macrophages. B. fragilis was shown to be phagocytized most efficiently in the presence of immune serum although normal mouse serum demonstrated reduced, but significant, opsonic activity. Phagocytosis was greater in the presence of serum collected from animals inoculated twice with the organism than in the presence of serum from once-inoculated animals. The increased opsonic activity of serum from twice-inoculated animals compared to singly inoculated animals was associated with increases in immunoglobulin G1(IgG1), IgG2a, and IgG2b but not IgM. Adsorption analysis of immune serum with homologous or heterologous bacterial antigens indicated that both antibody and complement act synergistically in opsonizing B. fragilis, although either alone may effectively opsonize this organism. Further evaluation of antibody-mediated opsonization revealed that prior treatment of heat-inactivated immune serum with the reducing agent 2-mercaptoethanol caused a slight, but significant, decrease in opsonic activity, thus indicating that IgM is a minor opsonizing antibody for B. fragilis. When ingestion of a B. fragilis stock strain (23745) was compared to a recent clinical isolate (C-1), it was observed that the stock strain was more easily phagocytized in the presence of normal mouse serum, thus suggesting a possible anti-opsonic-phagocytic property of the clinical strain. In addition, the clinical isolate was phagocytized to a significantly greater degree in an aerobic than an anaerobic environment. Subsequent analysis of in vitro killing of B. fragilis 23745 by peritoneal macrophages reflected the previous results in that optimal killing occurred in the presence of immune serum, although normal serum promoted phagocytic killing to an intermediate degree. Thus, these studies implicate both antibody and complement, either alone or in combination, in the opsonization of B. fragilis. Moreover, the virulence of clinical B. fragilis strains may relate to their refractoriness to opsonization and phagocytosis.

Full text

PDF
929

Selected References

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

  1. Bjornson A. B., Bjornson H. S., Kitko B. P. Participation of normal human immunoglobulins M, G, and A in opsonophagocytosis and intracellular killing of Bacteroides fragilis and Bacteroides thetaiotaomicron by human polymorphonuclear leukocytes. Infect Immun. 1980 May;28(2):633–637. doi: 10.1128/iai.28.2.633-637.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bjornson A. B., Bjornson H. S. Participation of immunoglobulin and the alternative complement pathway in opsonization of Bacteroides fragilis and Bacteroides thetaiotaomicron. J Infect Dis. 1978 Sep;138(3):351–358. doi: 10.1093/infdis/138.3.351. [DOI] [PubMed] [Google Scholar]
  3. Bjornson A. B., Michael J. G. Contribution of humoral and cellular factors to the resistance to experimental infection by Pseudomonas aeruginosa in mice. I. Interaction between immunoglobulins, heat-labile serum factors, and phagocytic cells in the killing of bacteria. Infect Immun. 1971 Oct;4(4):462–467. doi: 10.1128/iai.4.4.462-467.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. COHN Z. A., MORSE S. I. Interactions between rabbit polymorphonuclear leucocytes and staphylococci. J Exp Med. 1959 Sep 1;110:419–443. doi: 10.1084/jem.110.3.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casciato D. A., Rosenblatt J. E., Bluestone R., Goldberg L. S., Finegold S. M. Susceptibility of isolates of Bacteroides to the bactericidal activity of normal human serum. J Infect Dis. 1979 Jul;140(1):109–113. doi: 10.1093/infdis/140.1.109. [DOI] [PubMed] [Google Scholar]
  6. Dorrington K. J. Properties of the Fc receptor on macrophages. Immunol Commun. 1976;5(4):263–280. doi: 10.3109/08820137609044280. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. HIRSCH J. G., STRAUSS B. STUDIES ON HEAT-LABILE OPSONIN IN RABBIT SERUM. J Immunol. 1964 Jan;92:145–154. [PubMed] [Google Scholar]
  9. Hofstad T., Sveen K. Endotoxins of anaerobic gram-negative rods. Scand J Infect Dis Suppl. 1979;(19):42–45. [PubMed] [Google Scholar]
  10. Huber H., Fudenberg H. H. The interaction of monocytes and macrophages with immunoglobulins and complement. Ser Haematol. 1970;3(2):160–175. [PubMed] [Google Scholar]
  11. Ingham H. R., Sisson P. R., Tharagonnet D., Selkon J. B., Codd A. A. Inhibition of phagocytosis in vitro by obligate anaerobes. Lancet. 1977 Dec 17;2(8051):1252–1254. doi: 10.1016/s0140-6736(77)92662-9. [DOI] [PubMed] [Google Scholar]
  12. Kasper D. L., Hayes M. E., Reinap B. G., Craft F. O., Onderdonk A. B., Polk B. F. Isolation and identification of encapsulated strains of Bacteroides fragilis. J Infect Dis. 1977 Jul;136(1):75–81. doi: 10.1093/infdis/136.1.75. [DOI] [PubMed] [Google Scholar]
  13. Kasper D. L., Onderdonk A. B., Polk B. F., Bartlett J. G. Surface antigens as virulence factors in infection with Bacteroides fragilis. Rev Infect Dis. 1979 Mar-Apr;1(2):278–290. doi: 10.1093/clinids/1.2.278. [DOI] [PubMed] [Google Scholar]
  14. Kasper D. L., Onderdonk A. B., Reinap B. G., Linberg A. A. Variations of Bacteroides fragilis with in vitro passage: presence of an outer membrane-associated glycan and loss of capsular antigen. J Infect Dis. 1980 Nov;142(5):750–756. doi: 10.1093/infdis/142.5.750. [DOI] [PubMed] [Google Scholar]
  15. MUSCHEL L. H. Serum bactericidal actions. Ann N Y Acad Sci. 1960 Nov 21;88:1265–1272. doi: 10.1111/j.1749-6632.1960.tb20117.x. [DOI] [PubMed] [Google Scholar]
  16. MacLEOD C. M., KRAUS M. R. Relation of virulence of pneumococcal strains for mice to the quantity of capsular polysaccharide formed in vitro. J Exp Med. 1950 Jul 1;92(1):1–9. doi: 10.1084/jem.92.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. NELSON R. A., Jr, LEBRUN J. The requirement for antibody and complement for in vitro phagocytosis of starch granules. J Hyg (Lond) 1956 Mar;54(1):8–19. doi: 10.1017/s0022172400044259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Okuda K., Yanagi K., Takazoe I. Complement activation by Propionibacterium acnes and Bacteroides melaninogenicus. Arch Oral Biol. 1978;23(10):911–915. doi: 10.1016/0003-9969(78)90296-0. [DOI] [PubMed] [Google Scholar]
  19. Onderdonk A. B., Kasper D. L., Cisneros R. L., Bartlett J. G. The capsular polysaccharide of Bacteroides fragilis as a virulence factor: comparison of the pathogenic potential of encapsulated and unencapsulated strains. J Infect Dis. 1977 Jul;136(1):82–89. doi: 10.1093/infdis/136.1.82. [DOI] [PubMed] [Google Scholar]
  20. Quick J. D., Goldberg H. S., Sonnenwirth A. C. Human antibody to Bacteroidaceae. Am J Clin Nutr. 1972 Dec;25(12):1351–1356. doi: 10.1093/ajcn/25.12.1351. [DOI] [PubMed] [Google Scholar]
  21. Ralph P., Prichard J., Cohn M. Reticulum cell sarcoma: an effector cell in antibody-dependent cell-mediated immunity. J Immunol. 1975 Feb;114(2 Pt 2):898–905. [PubMed] [Google Scholar]
  22. Rhodes J. Receptor for monomeric IgM on guinea-pig splenic macrophages. Nature. 1973 Jun 29;243(5409):527–528. doi: 10.1038/243527a0. [DOI] [PubMed] [Google Scholar]
  23. Rowley D. Endotoxins and bacterial virulence. J Infect Dis. 1971 Mar;123(3):317–327. doi: 10.1093/infdis/123.3.317. [DOI] [PubMed] [Google Scholar]
  24. Sveen K. The importance of C5 and the role of the alternative complement pathway in leukocyte chemotaxis induced in vivo and in vitro by Bacteroides fragilis lipopolysaccharide. Acta Pathol Microbiol Scand B. 1978 Apr;86(2):93–100. doi: 10.1111/j.1699-0463.1978.tb00015.x. [DOI] [PubMed] [Google Scholar]
  25. Tizard I. R. Macrophage-cytophilic antibodies and the functions of macrophage-bound immunoglobulins. Bacteriol Rev. 1971 Dec;35(4):365–378. doi: 10.1128/br.35.4.365-378.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tofte R. W., Peterson P. K., Schmeling D., Bracke J., Kim Y., Quie P. G. Opsonization of four Bacteroides species: role of the classical complement pathway and immunoglobulin. Infect Immun. 1980 Mar;27(3):784–792. doi: 10.1128/iai.27.3.784-792.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Walker C. B., Wilkins T. D. Use of semisolid agar from initiation of pure Bacteroides fragilis infection in mice. Infect Immun. 1976 Sep;14(3):721–725. doi: 10.1128/iai.14.3.721-725.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES