Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1975 May 1;141(5):962–973. doi: 10.1084/jem.141.5.962

The in vitro induction of immunological tolerance in the B lymphocyte by oligovalent thymus-dependent antigens

PMCID: PMC2189784  PMID: 47897

Abstract

B-cell tolerance has been induced by oligovalent thymus-dependent antigens in an entirely in vitro system. Dissociated spleen cells from congenitally athymic (nu/nu) mice were preincubated for 24 h with 0.1 -- 1 mg/ml of either fowl gamma globulin (FGG) of DNP-human gamma globulin (DNP-HGG). After washing, the cells were tested for the ability to mount in vitro, thymus-independent responses against FGG and DNP. A state of specific responsiveness to either FGG or DNP was thus demonstrated. Features of this wholly in vitro system that paralleled previous findings on the in vivo induction of B-cell tolerance in nu/nu mice were the kinetics, 24 h being required for tolerance induction in either case, the abrogation of tolerance induction by the presence of POL both in vivo and in vitro, and finally the observation that in neither case was there a requirement for the antigens to be deaggregated. It was shown that DNP-(Fab) 2 fragments prepared from HGG induced DNP-specific tolerance indicating that the Fc piece was not required for tolerance induction in this in vitro system. DNP-bovine serum albumin was less effective than DNP-HGG or DNP-(Fab)2. Preincubation with subtoxic concentrations of DNP-lysine of DNP-epsilon- capric acid had only a marginal effect on DNP responsiveness. Since nu/nu mice, lacking in detectable T-cell function, were used as spleen cell donors, this work provides further evidence that B-cell tolerance to thymus-dependent antigens can be induced without the participation of T cells. It is suggested that B-cell tolerance to thymus-dependent antigens occurs when the antigen in a sufficient concentration and over a sufficient period of time has direct access to the B cell. This contact with antigen must be in the absence of an additional influence provided either by adjuvants like endotoxin or POL, or by activated macrophages, which may be stimulated by activated T cells; otherwise not tolerance but B-cell activation will occur.

Full Text

The Full Text of this article is available as a PDF (704.1 KB).

Selected References

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

  1. ADA G. L., NOSSAL G. J., PYE J. ANTIGENS IN IMMUNITY. III. DISTRIBUTION OF IODINATED ANTIGENS FOLLOWING INJECTION INTO RATS VIA THE HIND FOOTPADS. Aust J Exp Biol Med Sci. 1964 Jun;42:295–310. [PubMed] [Google Scholar]
  2. Baker P. J., Reed N. D., Stashak P. W., Amsbaugh D. F., Prescott B. Regulation of the antibody response to type 3 pneumococcal polysaccharide. I. Nature of regulatory cells. J Exp Med. 1973 Jun 1;137(6):1431–1441. doi: 10.1084/jem.137.6.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Basten A., Miller J. F., Sprent J., Pye J. A receptor for antibody on B lymphocytes. I. Method of detection and functional significance. J Exp Med. 1972 Mar 1;135(3):610–626. doi: 10.1084/jem.135.3.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britton S. Regulation of antibody synthesis against Escherichia coli endotoxin. IV. Induction of paralysis in vitro by treating normal lymphoid cells with antigen. J Exp Med. 1969 Mar 1;129(3):469–482. doi: 10.1084/jem.129.3.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Byers V. S., Sercarz E. E. Induction and reversal of immune paralysis in vitro. J Exp Med. 1970 Nov;132(5):845–857. doi: 10.1084/jem.132.5.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CLAMAN H. N. TOLERANCE TO A PROTEIN ANTIGEN IN ADULT MICE AND THE EFFECT OF NONSPECIFIC FACTORS. J Immunol. 1963 Dec;91:833–839. [PubMed] [Google Scholar]
  7. Cunningham A. J., Szenberg A. Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology. 1968 Apr;14(4):599–600. [PMC free article] [PubMed] [Google Scholar]
  8. Das S., Leskowitz S. The cellular basis for tolerance or immunity to bovine-gamma-globulin in mice. J Immunol. 1974 Jan;112(1):107–114. [PubMed] [Google Scholar]
  9. Diener E., Armstrong W. D. Immunological tolerance in vitro: kinetic studies at the cellular level. J Exp Med. 1969 Mar 1;129(3):591–603. doi: 10.1084/jem.129.3.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Diener E., Feldmann M. Relationship between antigen and antibody-induced suppression of immunity. Transplant Rev. 1972;8:76–103. doi: 10.1111/j.1600-065x.1972.tb01565.x. [DOI] [PubMed] [Google Scholar]
  11. Dresser D. W., Mitchison N. A. The mechanism of immunological paralysis. Adv Immunol. 1968;8:129–181. doi: 10.1016/s0065-2776(08)60466-6. [DOI] [PubMed] [Google Scholar]
  12. Feldmann M., Easten A. The relationship between antigenic structure and the requirement for thymus-derived cells in the immune response. J Exp Med. 1971 Jul 1;134(1):103–119. doi: 10.1084/jem.134.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feldmann M. Induction of immunity and tolerance in vitro by hapten protein conjugates. I. The relationship between the degree of hapten conjugation and the immunogenicity of dinitrophenylated polymerized flagellin. J Exp Med. 1972 Apr 1;135(4):735–753. doi: 10.1084/jem.135.4.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Feldmann M., Wagner H., Basten A., Holmes M. Humoral and cell mediated responses in vitro of spleen cells from mice with thymic aplasia (nude mice). Aust J Exp Biol Med Sci. 1972 Oct;50(5):651–660. doi: 10.1038/icb.1972.57. [DOI] [PubMed] [Google Scholar]
  15. Golub E. S., Weigle W. O. Studies on the induction of immunologic unresponsiveness. 3. Antigen form and mouse strain variation. J Immunol. 1969 Feb;102(2):389–396. [PubMed] [Google Scholar]
  16. Golub E. S., Weigle W. O. Studies on the induction of immunologic unresponsiveness. I. Effects of endotoxin and phytochemagglutinin. J Immunol. 1967 Jun;98(6):1241–1247. [PubMed] [Google Scholar]
  17. Katz D. H., Hamaoka T., Benacerraf B. Immunological tolerance in bone marrow-derived lymphocytes. 3. Tolerance induction in primed B cells by hapten conjugates of unrelated immunogenic or "nonimmunogenic" carriers. J Exp Med. 1974 Jun 1;139(6):1464–1472. doi: 10.1084/jem.139.6.1464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kindred B. Immunological unresponsiveness of genetically thymusless (nude) mice. Eur J Immunol. 1971 Jan;1(1):59–61. doi: 10.1002/eji.1830010114. [DOI] [PubMed] [Google Scholar]
  19. Kotlarski I., Courtenay B. M., Howard J. G. Studies on immunological paralysis. XI. A comparison of the tolerogenicity in vitro of levan and type 3 pneumococcal polysaccharide. Eur J Immunol. 1973 Aug;3(8):496–502. doi: 10.1002/eji.1830030809. [DOI] [PubMed] [Google Scholar]
  20. Louis J. A., Chiller J. M., Weigle W. O. The ability of bacterial lipopolysaccharide to modulate the induction of unresponsiveness to a state of immunity. Cellular parameters. J Exp Med. 1973 Dec 1;138(6):1481–1495. doi: 10.1084/jem.138.6.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Manning D. D., Reed N. D., Shaffer C. F. Maintenance of skin xenografts of widely divergent phylogenetic origin of congenitally athymic (nude) mice. J Exp Med. 1973 Aug 1;138(2):488–494. doi: 10.1084/jem.138.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller J. F., Warner N. L. The immune response of normal, irradiated and thymectomized mice to fowl immunoglobulin G as detected by a hemolytic plaque technique. Int Arch Allergy Appl Immunol. 1971;40(1):59–71. doi: 10.1159/000230395. [DOI] [PubMed] [Google Scholar]
  23. Mitchell G. F., Humphrey J. H., Wiliamson A. R. Inhibition of secondary anti-hapten responses with the hapten conjugated to type 3 pneumococcal polysaccharide. Eur J Immunol. 1972 Oct;2(5):460–467. doi: 10.1002/eji.1830020516. [DOI] [PubMed] [Google Scholar]
  24. Mitchison N. A. Immunological paralysis induced by brief exposure of cells to protein antigens. Immunology. 1968 Oct;15(4):531–547. [PMC free article] [PubMed] [Google Scholar]
  25. NISONOFF A., WISSLER F. C., LIPMAN L. N. Properties of the major component of a peptic digest of rabbit antibody. Science. 1960 Dec 9;132(3441):1770–1771. doi: 10.1126/science.132.3441.1770. [DOI] [PubMed] [Google Scholar]
  26. Osmond D. G., Nossal G. J. Differentiation of lymphocytes in mouse bone marrow. II. Kinetics of maturation and renewal of antiglobulin-binding cells studied by double labeling. Cell Immunol. 1974 Jul;13(1):132–145. doi: 10.1016/0008-8749(74)90233-0. [DOI] [PubMed] [Google Scholar]
  27. Schrader J. W. Induction of immunological tolerance to a thymus-dependent antigen in the absence of thymus-derived cells. J Exp Med. 1974 May 1;139(5):1303–1316. doi: 10.1084/jem.139.5.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schrader J. W. Mechanism of activation of the bone marrow-derived lymphocyte. 3. A distinction between a macrophage-produced triggering signal and the amplifying effect on triggered B lymphocytes of allogeneic interactions. J Exp Med. 1973 Dec 1;138(6):1466–1480. doi: 10.1084/jem.138.6.1466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schrader J. W., Nossal G. J. Effector cell blockade. A new mechanism of immune hyporeactivity induced by multivalent antigens. J Exp Med. 1974 Jun 1;139(6):1582–1598. doi: 10.1084/jem.139.6.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schrader J. W. Specific activation of the bone marrow-derived lymphocyte by antigen presented in a non-multivalent form. J Exp Med. 1973 Mar 1;137(3):844–849. doi: 10.1084/jem.137.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schrader J. W. The mechanism of bone marrow-derived (B) lymphocyte activation. I. Early events in antigen-induced triggering in the presence of polymerized flagellin. Eur J Immunol. 1974 Jan;4(1):14–20. doi: 10.1002/eji.1830040105. [DOI] [PubMed] [Google Scholar]
  32. Schrader J. W. Tolerance induction in B lymphocytes but thymus-dependent antigens. T cells may abrogate B-cell tolerance induction by prevent an antibody response. J Exp Med. 1975 May 1;141(5):974–989. doi: 10.1084/jem.141.5.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Talmage D. W., Pearlman D. S. The antibody response: a model based on antagonistic actions of antigen. J Theor Biol. 1963 Sep;5(2):321–339. doi: 10.1016/0022-5193(63)90067-5. [DOI] [PubMed] [Google Scholar]
  34. Watson J., Trenkner E., Cohn M. The use of bacterial lipopolysaccharides to show that two signals are required for the induction of antibody synthesis. J Exp Med. 1973 Sep 1;138(3):699–714. doi: 10.1084/jem.138.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wortis H. H. Immunological responses of 'nude' mice. Clin Exp Immunol. 1971 Feb;8(2):305–317. [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES