Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1983 Jun;40(3):1104–1111. doi: 10.1128/iai.40.3.1104-1111.1983

Polyclonal activation of human peripheral blood B lymphocytes by Fusobacterium nucleatum.

D F Mangan, D E Lopatin
PMCID: PMC348164  PMID: 6602093

Abstract

The present study examined in vitro polyclonal human B-lymphocyte (B-cell) activation (PBA) by Fusobacterium nucleatum. Pokeweed mitogen, a well-studied PBA activator, was included in some experiments for comparison. PBA was determined by the total immunoglobulin A, G, and M concentrations in the culture supernatants as measured by micro-enzyme-linked immunosorbent assay. F. nucleatum, at concentrations between 1 and 10 micrograms/ml, stimulated optimal PBA in monocyte-depleted cultures, whereas the pokeweed mitogen response was optimal in unfractionated, monocyte-containing cultures. Immunoglobulin synthesis occurred primarily between days 6 and 8 after stimulation with F. nucleatum. T lymphocytes enhanced the PBA response to F. nucleatum, particularly at a T- to B-cell ratio of 1:1. Immunoglobulin production was greater in round-bottomed wells than in flat-bottomed wells at lymphocyte concentrations of 200,000 cells per well. The PBA response, however, increased dramatically in flat-bottomed wells containing higher lymphocyte concentrations, suggesting that PBA is enhanced by cell-to-cell contact. A delay in stimulation of the lymphocytes with F. nucleatum resulted in diminished immunoglobulin production. The results provide information on the regulation of in vitro PBA induced by F. nucleatum. The data also suggest that there may be differences in the mechanisms by which F. nucleatum and pokeweed mitogen stimulate PBA.

Full text

PDF
1109

Selected References

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

  1. Allison A. C. Mechanisms by which activated macrophages inhibit lymphocyte responses. Immunol Rev. 1978;40:3–27. doi: 10.1111/j.1600-065x.1978.tb00399.x. [DOI] [PubMed] [Google Scholar]
  2. Alonso C. M., Bernabé R. R., Moreno E., Diaz de Espada F. Depletion of monocytes from human peripheral blood leucocytes by passage through sephadex G-10 columns. J Immunol Methods. 1978;22(3-4):361–368. doi: 10.1016/0022-1759(78)90043-1. [DOI] [PubMed] [Google Scholar]
  3. Bick P. H., Carpenter A. B., Holdeman L. V., Miller G. A., Ranney R. R., Palcanis K. G., Tew J. G. Polyclonal B-cell activation induced by extracts of Gram-negative bacteria isolated from periodontally diseased sites. Infect Immun. 1981 Oct;34(1):43–49. doi: 10.1128/iai.34.1.43-49.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  5. Chen P., Trummel C., Horton J., Baker J. J., Oppenheim J. J. Production of osteoclast-activating factor by normal human peripheral blood rosetting and nonrosetting lymphocytes. Eur J Immunol. 1976 Oct;6(10):732–736. doi: 10.1002/eji.1830061014. [DOI] [PubMed] [Google Scholar]
  6. Clagett J. A., Engel D. Polyclonal activation: a form of primitive immunity and its possible role in pathogenesis of inflammatory diseases. Dev Comp Immunol. 1978 Apr;2(2):235–241. doi: 10.1016/s0145-305x(78)80066-4. [DOI] [PubMed] [Google Scholar]
  7. Fauci A. S., Whalen G., Burch C. Activation of human B lymphocytes XVI. Cellular requirements, interactions, and immunoregulation of pokeweed mitogen-induced total-immunoglobulin producing plaque-forming cells in peripheral blood. Cell Immunol. 1980 Aug 15;54(1):230–240. doi: 10.1016/0008-8749(80)90204-x. [DOI] [PubMed] [Google Scholar]
  8. Gerrard T. L., Fauci A. S. Activation and immunoregulation of antigen-specific human b lymphocyte responses: multifaceted role of the monocyte. J Immunol. 1982 May;128(5):2367–2372. [PubMed] [Google Scholar]
  9. Kaplan M. E., Clark C. An improved rosetting assay for detection of human T lymphocytes. J Immunol Methods. 1974 Jul;5(2):131–135. doi: 10.1016/0022-1759(74)90003-9. [DOI] [PubMed] [Google Scholar]
  10. Kunori T., Ringdén O. Antibody secretion and DNA synthesis induced by lipopolysaccharide in enriched human lymphocyte subpopulations. Scand J Immunol. 1980;11(2):137–145. doi: 10.1111/j.1365-3083.1980.tb00219.x. [DOI] [PubMed] [Google Scholar]
  11. Lally E. T., Baehni P. C., McArthur W. P. Local immunoglobulin synthesis in periodontal disease. J Periodontal Res. 1980 Mar;15(2):159–164. doi: 10.1111/j.1600-0765.1980.tb00270.x. [DOI] [PubMed] [Google Scholar]
  12. Lane H. C., Volkman D. J., Whalen G., Fauci A. S. In vitro antigen-induced, antigen-specific antibody production in man. Specific and polyclonal components, kinetics, and cellular requirements. J Exp Med. 1981 Oct 1;154(4):1043–1057. doi: 10.1084/jem.154.4.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lipsky P. E. Staphylococcal protein A, a T cell-regulated polyclonal activator of human B cells. J Immunol. 1980 Jul;125(1):155–162. [PubMed] [Google Scholar]
  14. Lopatin D. E., Mangan D. F., Horner I. S., Peebles F. L. Mitogen-induced amplification of blastogenesis in lipopolysaccharide-precultured lymphocytes. Infect Immun. 1980 Aug;29(2):512–519. doi: 10.1128/iai.29.2.512-519.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lovelace B. M., 3rd, Thompson J. J., Yukna R. A. Evidence for local immunoglobulin synthesis in periodontitis. J Periodontol. 1982 Oct;53(10):626–630. doi: 10.1902/jop.1982.53.10.626. [DOI] [PubMed] [Google Scholar]
  16. Lêthibichthuy, Ciorbaru R., Brochier J. Human B cell differentiation. I. Immunoglobulin synthesis induced by Nocardia mitogen. Eur J Immunol. 1978 Feb;8(2):119–123. doi: 10.1002/eji.1830080209. [DOI] [PubMed] [Google Scholar]
  17. Mackler B. F., Altman L. C., Wahl S., Rosenstreich D. L., Oppenheim J. J., Mergenhagen S. E. Blastogenesis and lymphokine synthesis by T and B lymphocytes from patients with periodontal disease. Infect Immun. 1974 Oct;10(4):844–850. doi: 10.1128/iai.10.4.844-850.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mangan D. F., Lopatin D. E. In vitro stimulation of immunoglobulin production from human peripheral blood lymphocytes by a soluble preparation of Actinomyces viscosus. Infect Immun. 1981 Jan;31(1):236–244. doi: 10.1128/iai.31.1.236-244.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Montazeri G., Chiorazzi N., Fu S. M., Kunkel H. G. Regulatory role of circulating monocytes in the differentiative and proliferative responses of human B lymphocytes. Clin Immunol Immunopathol. 1980 May;16(1):1–10. doi: 10.1016/0090-1229(80)90160-9. [DOI] [PubMed] [Google Scholar]
  20. Munoz J., Pryjma J., Fudenberg H. H., Virella G. Immunoglobulin quantitation and enumeration of immunoglobulin-producing cells: comparison of two index of B-cell activation. Scand J Immunol. 1980;12(4):345–353. doi: 10.1111/j.1365-3083.1980.tb00076.x. [DOI] [PubMed] [Google Scholar]
  21. Newman M. G., Socransky S. S., Savitt E. D., Propas D. A., Crawford A. Studies of the microbiology of periodontosis. J Periodontol. 1976 Jul;47(7):373–379. doi: 10.1902/jop.1976.47.7.373. [DOI] [PubMed] [Google Scholar]
  22. Nishikawa S. I., Hirata T., Nagai T., Mayumi M., Izumi T. PPD-induced immunoglobulin production in human peripheral blood lymphocytes. I. Necessary conditions for inducing the response. J Immunol. 1979 Jun;122(6):2143–2149. [PubMed] [Google Scholar]
  23. Philp J. R., Huffman A. L., Johnson J. E., 3rd Amplified migration inhibition effect. Infect Immun. 1976 Oct;14(4):872–875. doi: 10.1128/iai.14.4.872-875.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pryjma J., Muñoz J., Galbraith R. M., Fudenberg H. H., Virella G. Induction and suppression of immunoglobulin synthesis in cultures of human lymphocytes: effects of pokeweed mitogen and Staphylococcus aureus Cowan I. J Immunol. 1980 Feb;124(2):656–661. [PubMed] [Google Scholar]
  25. Rosenberg S. A., Lipsky P. E. The role of monocyte factors in the differentiation of immunoglobulin secreting cells from human peripheral blood B cells. J Immunol. 1980 Jul;125(1):232–237. [PubMed] [Google Scholar]
  26. Rosenberg S. A., Lipsky P. E. The role of monocytes in pokeweed mitogen-stimulated human B cell activation: separate requirements for intact monocytes and a soluble monocyte factor. J Immunol. 1981 Apr;126(4):1341–1345. [PubMed] [Google Scholar]
  27. Rümke H. C., Terpstra F. G., Out T. A., Vossen J. M., Zeijlemaker W. P. Immunoglobulin production by human lymphocytes in a microculture system: culture conditions and cellular interactions. Clin Immunol Immunopathol. 1981 Jun;19(3):338–350. doi: 10.1016/0090-1229(81)90077-5. [DOI] [PubMed] [Google Scholar]
  28. Sabiston C. B., Jr, Gold W. A. Anaerobic bacteria in oral infections. Oral Surg Oral Med Oral Pathol. 1974 Aug;38(2):187–192. doi: 10.1016/0030-4220(74)90054-1. [DOI] [PubMed] [Google Scholar]
  29. Seymour G. J., Greenspan J. S. The phenotypic characterization of lymphocyte subpopulations in established human periodontal disease. J Periodontal Res. 1979 Jan;14(1):39–46. doi: 10.1111/j.1600-0765.1979.tb00216.x. [DOI] [PubMed] [Google Scholar]
  30. Slots J., Möenbo D., Langebaek J., Frandsen A. Microbiota of gingivitis in man. Scand J Dent Res. 1978 May;86(3):174–181. doi: 10.1111/j.1600-0722.1978.tb01929.x. [DOI] [PubMed] [Google Scholar]
  31. Slots J. The predominant cultivable microflora of advanced periodontitis. Scand J Dent Res. 1977 Jan-Feb;85(2):114–121. doi: 10.1111/j.1600-0722.1977.tb00541.x. [DOI] [PubMed] [Google Scholar]
  32. Smith S., Bick P. H., Miller G. A., Ranney R. R., Rice P. L., Lalor J. H., Tew J. G. Polyclonal B-cell activation: severe periodontal disease in young adults. Clin Immunol Immunopathol. 1980 Jul;16(3):354–366. doi: 10.1016/0090-1229(80)90141-5. [DOI] [PubMed] [Google Scholar]
  33. Volkman D. J., Allyn S. P., Fauci A. S. Antigen-induced in vitro antibody production in humans: tetanus toxoid-specific antibody synthesis. J Immunol. 1982 Jul;129(1):107–112. [PubMed] [Google Scholar]
  34. Volkman D. J., Lane H. C., Fauci A. S. Antigen-induced in vitro antibody production in humans: a model for B cell activation and immunoregulation. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2528–2531. doi: 10.1073/pnas.78.4.2528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Williams B. L., Pantalone R. M., Sherris J. C. Subgingival microflora and periodontitis. J Periodontal Res. 1976 Feb;11(1):1–18. doi: 10.1111/j.1600-0765.1976.tb00045.x. [DOI] [PubMed] [Google Scholar]
  36. Wittgow W. C., Jr, Sabiston C. B., Jr Microorganisms from pulpal chambers of intact teeth with necrotic pulps. J Endod. 1975 May;1(5):168–171. doi: 10.1016/S0099-2399(75)80115-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES