Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1979 Sep 19;150(3):491–506. doi: 10.1084/jem.150.3.491

The induction of hapten-specific immunological tolerance and immunity in B lymphocytes. VI. Differential tolerance susceptibility in adult spleen as a function of B-cell maturation level

PMCID: PMC2185647  PMID: 158060

Abstract

The maturation level of the B-lymphocyte subpopulations involved in trinitrophenyl (TNP)-specific immunological tolerance in adult mice induced by the injection of trinitrobenzenesulfonic acid (TNBS) was investigated using in vitro antigen-specific and nonspecific polyclonal stimulation. The maturity of the B-cell subsets being studied was defined by the antigen or polyclonal activator which evoked a response. Thus, the thymic independent (TI-1) antigen TNP-lipopolysaccharide (TNP- LPS) and the polyclonal stimulant LPS were used to activate immature, neonatal-type B lymphocytes, whereas mature, adult-type B cells were responsive to the TI-2 antigen, TNP-Ficoll, and the nonspecific activator, purified protein derivative (PPD). Whereas unresponsiveness in TNP-LPS-reactive (immature) B cells 4 d after TNBS treatment was previously shown to be the result of functional deletion, partially reversible receptor blockade was detected in this study early after tolerogen treatment. By the 24-h point, tolerance was irreversible, as assessed by 24-h of antigen-free incubation and cocultivation of tolerant cells with control splenocytes. Tolerance was induced more rapidly in immature, TI-1 B cells than in mature TI-2 B lymphocytes. B lymphocytes reactive to TNP-Ficoll were also less susceptible to receptor blockade. Using LPS as a nonspecific probe for immature B cells, 60% tolerance in high affinity TNP-specific cells was induced within 12 h of TNBS treatment, and complete unresponsiveness by 24 h. In contrast, no significant decrease in response to the mature B-cell activator, PPD, occurred until day 2. Furthermore, the 50% tolerance level was achieved in TNP-specific LPS-reactive B cells by 100 times less tolerogen than required for PPD-responsive cells. Thus, TNBS- induced unresponsiveness in cells reactive to TNP-LPS is initially a result of reversible receptor blockade which leads within 4 d to functional deletion. Immature, TI-1 B lymphocytes, which give polyclonal responses to LPS and antigen-specific responses to TNP-LPS, are rendered tolerant to TNBS more rapidly and at lower tolerogen does than mature, TI-2 mouse B cells which react polyclonally to PPD and specifically to TNP-Ficoll. Moreover, these data show that both the immature and the mature B lymphocyts with these characteristic tolerance susceptibilities and specific and nonspecific immune response patterns are present in the adult mouse spleen.

Full Text

The Full Text of this article is available as a PDF (918.4 KB).

Selected References

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

  1. Ahmed A., Scher I., Sharrow S. O., Smith A. H., Paul W. E., Sachs D. H., Sell K. W. B-lymphocyte heterogeneity: development and characterization of an alloantiserum which distinguishes B-lymphocyte differentiation alloantigens. J Exp Med. 1977 Jan 1;145(1):101–110. doi: 10.1084/jem.145.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aldo-Benson M., Borel Y. Loss of carrier-determined tolerance in vitro with loss of receptor blockade. J Immunol. 1976 Jan;116(1):223–226. [PubMed] [Google Scholar]
  3. Amsbaugh D. F., Hansen C. T., Prescott B., Stashak P. W., Barthold D. R., Baker P. J. Genetic control of the antibody response to type 3 pneumococcal polysaccharide in mice. I. Evidence that an X-linked gene plays a decisive role in determining responsiveness. J Exp Med. 1972 Oct 1;136(4):931–949. doi: 10.1084/jem.136.4.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andersson B., Wigzell H. Studies on antibody avidity at the cellular level. Effects of immunological paralysis and administered antibody. Eur J Immunol. 1971 Nov;1(5):384–390. doi: 10.1002/eji.1830010516. [DOI] [PubMed] [Google Scholar]
  5. Bruyns C., Urbain-Vansanten G., Planard C., Vos-Cloetens C., Urbain J. Ontogeny of mouse B lymphocytes and inactivation by antigen of early B lymphocytes. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2462–2466. doi: 10.1073/pnas.73.7.2462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cambier J. C., Kettman J. R., Vitetta E. S., Uhr J. W. Differential susceptibility of neonatal and adult murine spleen cells to in vitro induction of B-cell tolerance. J Exp Med. 1976 Jul 1;144(1):293–297. doi: 10.1084/jem.144.1.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cambier J. C., Vitetta E. S., Kettman J. R., Wetzel G. M., Uhr J. W. B-cell tolerance. III. Effect of papain-mediated cleavage of cell surface IgD on tolerance susceptibility of murine B cells. J Exp Med. 1977 Jul 1;146(1):107–117. doi: 10.1084/jem.146.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cantor H., Boyse E. Regulation of the immune response by T-cell subclasses. Contemp Top Immunobiol. 1977;7:47–67. doi: 10.1007/978-1-4684-3054-7_2. [DOI] [PubMed] [Google Scholar]
  9. Coutinho A., Möller G. Editorial: Immune activation of B cells: evidence for 'one nonspecific triggering signal' not delivered by the Ig receptors. Scand J Immunol. 1974;3(2):133–146. [PubMed] [Google Scholar]
  10. Coutinho A., Möller G. The self-nonself discrimination: A one-signal mechanism. Scand J Immunol. 1975;4(2):99–102. doi: 10.1111/j.1365-3083.1975.tb02605.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Davie J. M., Paul W. E., Katz D. H., Benacerraf B. Hapten-specific tolerance. Preferential depression of the high affinity antibody response. J Exp Med. 1972 Sep 1;136(3):426–438. doi: 10.1084/jem.136.3.426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fidler J. M., Golub E. S. Immunologic tolerance to a hapten. 3. Induction of tolerance to trinitrophenyl in B cells in various differentiation states. J Immunol. 1974 May;112(5):1891–1899. [PubMed] [Google Scholar]
  14. Fidler J. M., Golub E. S. Immunological tolerance to a hapten. I. Induction and maintenance of tolerance to trinitrophenyl with trinitrobenzene sulfonic acid. J Exp Med. 1973 Jan 1;137(1):42–54. doi: 10.1084/jem.137.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fidler J. M., Golub E. S. Induction of tolerance to a hapten. II. Maintenance and escape of the tolerant state to TNP. J Immunol. 1973 Aug;111(2):317–323. [PubMed] [Google Scholar]
  16. Fidler J. M. In vivo immune response to TNP hapten coupled to thymus-independent carrier lipopolysaccharide. Cell Immunol. 1975 Apr;16(2):223–236. doi: 10.1016/0008-8749(75)90114-8. [DOI] [PubMed] [Google Scholar]
  17. Fidler J. M. The induction of hapten-specific immunologic tolerance and immunity in B lymphocytes. III. In vitro evaluation of TNP-specific tolerance and evidence for B cell deletion. J Immunol. 1978 Jul;121(1):245–254. [PubMed] [Google Scholar]
  18. Fidler J. M. The induction of hapten-specific immunologic tolerance and immunity in B lymphocytes. IV. Induction of TNP-specific unresponsiveness in vitro with serum from tolerant mice. J Immunol. 1978 Oct;121(4):1558–1565. [PubMed] [Google Scholar]
  19. Fidler J. M. The induction of hapten-specific immunological tolerance and immunity in B lymphocytes. II. Temporal and dose response relationships of tolerance induction in B lymphocytes of various differentiation states. Cell Immunol. 1976 May;23(2):240–253. doi: 10.1016/0008-8749(76)90190-8. [DOI] [PubMed] [Google Scholar]
  20. Finkelman F. D., Smith A. H., Scher I., Paul W. E. Abnormal ratio of membrane immunoglobulin classes in mice with an X-linked B-lymphocyte defect. J Exp Med. 1975 Nov 1;142(5):1316–1321. doi: 10.1084/jem.142.5.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Goodman M. G., Fidler J. M., Weigle W. O. Nonspecific activation of murine lymphocytes. IV. Proliferation of a distinct, late maturing lymphocyte subpopulation induced by 2-mercaptoethanol. J Immunol. 1978 Nov;121(5):1905–1913. [PubMed] [Google Scholar]
  22. Gronowicz E., Coutinho A. Hapten-induced B cell paralysis. II. Evidence for trivial mechanisms of tolerance. Eur J Immunol. 1976 Jun;5(6):413–420. doi: 10.1002/eji.1830050611. [DOI] [PubMed] [Google Scholar]
  23. Gronowicz E., Coutinho A., Möller G. Differentiation of B cells: sequential appearance of responsiveness to polyclonal activators. Scand J Immunol. 1974;3(4):413–421. doi: 10.1111/j.1365-3083.1974.tb01274.x. [DOI] [PubMed] [Google Scholar]
  24. Inman J. K. Thymus-independent antigens: the preparation of covalent, hapten-ficoll conjugates. J Immunol. 1975 Feb;114(2 Pt 1):704–709. [PubMed] [Google Scholar]
  25. Kearney J. F., Cooper M. D., Klein J., Abney E. R., Parkhouse R. M., Lawton A. R. Ontogeny of Ia and IgD on IgM-bearing B lymphocytes in mice. J Exp Med. 1977 Jul 1;146(1):297–301. doi: 10.1084/jem.146.1.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kearney J. F., Cooper M. D., Lawton A. R. B lymphocyte differentiation induced by lipopolysaccharide. III. Suppression of B cell maturation by anti-mouse immunoglobulin antibodies. J Immunol. 1976 Jun;116(6):1664–1668. [PubMed] [Google Scholar]
  27. Kettman J., Dutton R. W. An in vitro primary immune response to 2,4,6-trinitrophenyl substituted erythrocytes: response against carrier and hapten. J Immunol. 1970 Jun;104(6):1558–1561. [PubMed] [Google Scholar]
  28. Merchant B., Snippe H., Lizzio E. F., Inman J. K. Cellular and molecular requirements for X-linked, hapten-specific B-cell blockade in CBA/N mice. J Exp Med. 1978 Jun 1;147(6):1755–1770. doi: 10.1084/jem.147.6.1755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Merchant B., Snippe H., Lizzio E. F., Inman J. K. X-linked genetic control of hapten-polysaccharide-mediated specific immune unresponsiveness in CBA/N mice. J Immunol. 1978 Apr;120(4):1362–1368. [PubMed] [Google Scholar]
  30. Metcalf E. S., Klinman N. R. In vitro tolerance induction of neonatal murine B cells. J Exp Med. 1976 Jun 1;143(6):1327–1340. doi: 10.1084/jem.143.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miller J. F. Lymphocyte interactions in antibody responses. Int Rev Cytol. 1972;33:77–130. doi: 10.1016/s0074-7696(08)61449-7. [DOI] [PubMed] [Google Scholar]
  32. Mosier D. E., Johnson B. M. Ontogeny of mouse lymphocyte function. II. Development of the ability to produce antibody is modulated by T lymphocytes. J Exp Med. 1975 Jan 1;141(1):216–226. doi: 10.1084/jem.141.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Raff M. C., Owen J. J., Cooper M. D., Lawton A. R., 3rd, Megson M., Gathings W. E. Differences in susceptibility of mature and immature mouse B lymphocytes to anti-immunoglobulin-induced immunoglobulin suppression in vitro. Possible implications for B-cell tolerance to self. J Exp Med. 1975 Nov 1;142(5):1052–1064. doi: 10.1084/jem.142.5.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rittenberg M. B., Pratt K. L. Antitrinitrophenyl (TNP) plaque assay. Primary response of Balb/c mice to soluble and particulate immunogen. Proc Soc Exp Biol Med. 1969 Nov;132(2):575–581. doi: 10.3181/00379727-132-34264. [DOI] [PubMed] [Google Scholar]
  35. Rosenberg Y. J., Parish C. R. Ontogeny of the antibody-forming cell line in mice. IV. Appearance of cells bearing Fc receptors, complement receptors, and surface immunoglobulin. J Immunol. 1977 Feb;118(2):612–617. [PubMed] [Google Scholar]
  36. Scher I., Sharrow S. O., Paul W. E. X-linked B-lymphocyte defect in CBA/N mice. III. Abnormal development of B-lymphocyte populations defined by their density of surface immunoglobulin. J Exp Med. 1976 Aug 1;144(2):507–518. doi: 10.1084/jem.144.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schlegel R. A. Antigen-initiated B lymphocyte differentiation. Kinetics and T lymphocyte dependence of the primary and secondary in vitro immune responses to the hapten 4-hydroxy-3-iodo-5-nitrophenylacetic acid presented on the carrier polymerised bacterial flagellin. Aust J Exp Biol Med Sci. 1974 Jun;52(Pt 3):471–489. [PubMed] [Google Scholar]
  38. Scott D. W., Layton J. E., Nossal G. J. Role of IgD in the immune response and tolerance. I. Anti-delta pretreatment facilitates tolerance induction in adult B cells in vitro. J Exp Med. 1977 Dec 1;146(6):1473–1483. doi: 10.1084/jem.146.6.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shortman K., Williams N., Adams P. The separation of different cell classes from lymphoid organs. V. Simple procedures for the removal of cell debris. Damaged cells and erythroid cells from lymphoid cell suspensions. J Immunol Methods. 1972 May;1(3):273–287. doi: 10.1016/0022-1759(72)90005-1. [DOI] [PubMed] [Google Scholar]
  40. Sidman C. L., Unanue E. R. Development of B lymphocytes. I. Cell populations and a critical event during ontogeny. J Immunol. 1975 Jun;114(6):1730–1735. [PubMed] [Google Scholar]
  41. Sidman C. L., Unanue E. R. Receptor-mediated inactivation of early B lymphocytes. Nature. 1975 Sep 11;257(5522):149–151. doi: 10.1038/257149a0. [DOI] [PubMed] [Google Scholar]
  42. Siskind G. W., Benacerraf B. Cell selection by antigen in the immune response. Adv Immunol. 1969;10:1–50. doi: 10.1016/s0065-2776(08)60414-9. [DOI] [PubMed] [Google Scholar]
  43. Stocker J. W. Tolerance induction in maturing B cells. Immunology. 1977 Mar;32(3):283–290. [PMC free article] [PubMed] [Google Scholar]
  44. Theis G. A., Siskind G. W. Selection of cell populations in induction of tolerance: affinity of antibody formed in partially tolerant rabbits. J Immunol. 1968 Jan;100(1):138–141. [PubMed] [Google Scholar]
  45. Vitetta E. S., Cambier J. C., Ligler F. S., Kettman J. R., Uhr J. W. B-cell tolerance. IV. Differential role of surface IgM and IgD in determining tolerance susceptibility of murine B cells. J Exp Med. 1977 Dec 1;146(6):1804–1808. doi: 10.1084/jem.146.6.1804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vitetta E. S., Melcher U., McWilliams M., Lamm M. E., Phillips-Quagliata J. M., Uhr J. W. Cell surface immunoglobulin. XI. The appearance of an IgD-like molecule on murine lymphoid cells during ontogeny. J Exp Med. 1975 Jan 1;141(1):206–215. doi: 10.1084/jem.141.1.206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vitetta E. S., Uhr J. W. Immunoglobulin-receptors revisited. Science. 1975 Sep 19;189(4207):964–969. doi: 10.1126/science.1083069. [DOI] [PubMed] [Google Scholar]
  48. Weigle W. O. Immunological unresponsiveness. Adv Immunol. 1973;16:61–122. doi: 10.1016/s0065-2776(08)60296-5. [DOI] [PubMed] [Google Scholar]
  49. Williams N., Kraft N., Shortman K. The separation of different cell classes from lymphoid organs. VI. The effect of osmolarity of gradient media on the density distribution of cells. Immunology. 1972 May;22(5):885–899. [PMC free article] [PubMed] [Google Scholar]
  50. Zitron I. M., Mosier D. E., Paul W. E. The role of surface IgD in the response to thymic-independent antigens. J Exp Med. 1977 Dec 1;146(6):1707–1718. doi: 10.1084/jem.146.6.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES