Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Evidence from the generation of immunoglobulin G–secreting cells that stochastic mechanisms regulate lymphocyte differentiation

Abstract

Naive B lymphocytes undergo isotype switching and develop into immunoglobulin-secreting cells to generate the appropriate class and amount of antibody necessary for effective immunity. Although this seems complex, we report here that the generation of immunoglobulin G–secreting cells from naive precursors is highly predictable. The probabilities of isotype switching and development into secreting cells change with successive cell divisions and interleave independently. Cytokines alter the probability of each differentiation event, while leaving intact their independent assortment. As a result, cellular heterogeneity arises automatically as the cells divide. Stochastic division-linked regulation of heterogeneity challenges the conventional paradigms linking distinct phenotypes to unique combinations of signals and has the potential to simplify our concept of immune complexity considerably.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: IL-4-induced immunoglobulin secretion shows a cell division–related increase in ASC numbers.
Figure 2: Identification of immunoglobulin-producing B cells by flow cytometry.
Figure 3: Tracking simultaneous immunoglobulin switching and development of ASCs.
Figure 4: IL-5 reduces switching and increases the appearance of Synd1+ cells in each division.
Figure 5: ASCs downregulate division-associated isotype switching.
Figure 6: Confirmation of division-associated increases in differentiation.
Figure 7: The generation of cellular heterogeneity by two independent differentiation events.

Similar content being viewed by others

References

  1. Benner, R., Hijmans, W. & Haaijman, J.J. The bone marrow: the major source of serum immunoglobulins, but still a neglected site of antibody formation. Clin. Exp. Immunol. 46, 1–8 (1981).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. van Rooijen, N., Claassen, E. & Eiekelenboom, P. Is there a single differentiation pathway for all antibody-forming cells in the spleen? Immunol. Today 7, 193–195 (1986).

    Article  CAS  Google Scholar 

  3. Jacob, J., Kassir, R. & Kelsoe, G. In situ studies of the primary immune response to (4-hydroxy-3- nitrophenyl)acetyl. I. The architecture and dynamics of responding cell populations. J. Exp. Med. 173, 1165–1175 (1991).

    Article  CAS  Google Scholar 

  4. Smith, K.G., Hewitson, T.D., Nossal, G.J. & Tarlinton, D.M. The phenotype and fate of the antibody-forming cells of the splenic foci. Eur. J. Immunol. 26, 444–448 (1996).

    Article  CAS  Google Scholar 

  5. Smith, K.G., Light, A., Nossal, G.J. & Tarlinton, D.M. The extent of affinity maturation differs between the memory and antibody-forming cell compartments in the primary immune response. EMBO J. 16, 2996–3006 (1997).

    Article  CAS  Google Scholar 

  6. Takahashi, Y., Dutta, P.R., Cerasoli, D.M. & Kelsoe, G. In situ studies of the primary immune response to (4-hydroxy-3- nitrophenyl)acetyl. V. Affinity maturation develops in two stages of clonal selection. J. Exp. Med. 187, 885–895 (1998).

    Article  CAS  Google Scholar 

  7. Szakal, A.K., Kosco, M.H. & Tew, J.G. Microanatomy of lymphoid tissue during humoral immune responses: structure function relationships. Annu. Rev. Immunol. 7, 91–109 (1989).

    Article  CAS  Google Scholar 

  8. Ochsenbein, A.F. et al. Protective long-term antibody memory by antigen-driven and T help-dependent differentiation of long-lived memory B cells to short-lived plasma cells independent of secondary lymphoid organs. Proc. Natl. Acad. Sci. USA 97, 13263–13268 (2000).

    Article  CAS  Google Scholar 

  9. Ho, F., Lortan, J.E., MacLennan, I.C. & Khan, M. Distinct short-lived and long-lived antibody-producing cell populations. Eur. J. Immunol. 16, 1297–1301 (1986).

    Article  CAS  Google Scholar 

  10. Slifka, M.K., Antia, R., Whitmire, J.K. & Ahmed, R. Humoral immunity due to long-lived plasma cells. Immunity 8, 363–372 (1998).

    Article  CAS  Google Scholar 

  11. Manz, R.A., Lohning, M., Cassese, G., Thiel, A. & Radbruch, A. Survival of long-lived plasma cells is independent of antigen. Int. Immunol. 10, 1703–1711 (1998).

    Article  CAS  Google Scholar 

  12. Lalor, P.A., Nossal, G.J., Sanderson, R.D. & McHeyzer-Williams, M.G. Functional and molecular characterization of single, (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific, IgG1+ B cells from antibody-secreting and memory B cell pathways in the C57BL/6 immune response to NP. Eur. J. Immunol. 22, 3001–3011 (1992).

    Article  CAS  Google Scholar 

  13. Kopper, L. & Sebestyen, A. Syndecans and the lymphoid system. Leuk. Lymphoma 38, 271–281 (2000).

    Article  CAS  Google Scholar 

  14. Hodgkin, P.D. & Kehry, M.R. Methods for polyclonal B lymphocyte activation to proliferation and Ig secretion in vitro. in Weir's Handbook of Experimental Immunology, Vol. 89, 89.1–89.13 (eds. Herzenberg, L.A., Weir, D.M., Herzenberg, L.A. & Blackwell, C.) (Blackwell Science, Cambridge, Massachusetts, 1996).

    Google Scholar 

  15. Hodgkin, P.D., Lee, J.H. & Lyons, A.B. B cell differentiation and isotype switching is related to division cycle number. J. Exp. Med. 184, 277–281 (1996).

    Article  CAS  Google Scholar 

  16. Hasbold, J., Lyons, A.B., Kehry, M.R. & Hodgkin, P.D. Cell division number regulates IgG1 and IgE switching of B cells following stimulation by CD40 ligand and IL-4. Eur. J. Immunol. 28, 1040–1051 (1998).

    Article  CAS  Google Scholar 

  17. Hasbold, J., Hong, J.S., Kehry, M.R. & Hodgkin, P.D. Integrating signals from IFN-γ and IL-4 by B cells: positive and negative effects on CD40 ligand-induced proliferation, survival, and division-linked isotype switching to IgG1, IgE, and IgG2a. J. Immunol. 163, 4175–4181 (1999).

    CAS  PubMed  Google Scholar 

  18. Deenick, E.K., Hasbold, J. & Hodgkin, P.D. Switching to IgG3, IgG2b, and IgA is division linked and independent, revealing a stochastic framework for describing differentiation. J. Immunol. 163, 4707–4714 (1999).

    CAS  PubMed  Google Scholar 

  19. Tangye, S.G., Ferguson, A., Avery, D.T., Ma, C.S. & Hodgkin, P.D. Isotype switching by human B cells is division-associated and regulated by cytokines. J. Immunol. 169, 4298–4306 (2002).

    Article  CAS  Google Scholar 

  20. Janas, M.L., Hodgkin, P., Hibbs, M. & Tarlinton, D. Genetic evidence for Lyn as a negative regulator of IL-4 signaling. J. Immunol. 163, 4192–4198 (1999).

    CAS  PubMed  Google Scholar 

  21. Maliszewski, C.R. et al. Recombinant CD40 ligand stimulation of murine B cell growth and differentiation: cooperative effects of cytokines. Eur. J. Immunol. 23, 1044–1049 (1993).

    Article  CAS  Google Scholar 

  22. Hollenbaugh, D., Ochs, H.D., Noelle, R.J., Ledbetter, J.A. & Aruffo, A. The role of CD40 and its ligand in the regulation of the immune response. Immunol. Rev. 138, 23–37 (1994).

    Article  CAS  Google Scholar 

  23. McIntyre, T.M., Kehry, M.R. & Snapper, C.M. Novel in vitro model for high-rate IgA class switching. J. Immunol. 154, 3156–3161 (1995).

    CAS  PubMed  Google Scholar 

  24. Calame, K.L. Plasma cells: finding new light at the end of B cell development. Nat. Immunol. 2, 1103–1108 (2001).

    Article  CAS  Google Scholar 

  25. Jelinek, D.F. & Lipsky, P.E. The role of B cell proliferation in the generation of immunoglobulin-secreting cells in man. J. Immunol. 130, 2597–2604 (1983).

    CAS  PubMed  Google Scholar 

  26. Jego, G. et al. Reactive plasmacytoses are expansions of plasmablasts retaining the capacity to differentiate into plasma cells. Blood 94, 701–712 (1999).

    CAS  PubMed  Google Scholar 

  27. Sze, D.M., Toellner, K.M., Garcia de Vinuesa, C., Taylor, D.R. & MacLennan, I.C. Intrinsic constraint on plasmablast growth and extrinsic limits of plasma cell survival. J. Exp. Med. 192, 813–821 (2000).

    Article  CAS  Google Scholar 

  28. Tangye, S., Avery, D. & Hodgkin, P. A division-linked mechanism for the rapid generation of Ig-secreting cells from human memory B cells. J. Immunol. 170, 261–269 (2003).

    Article  CAS  Google Scholar 

  29. Tangye, S., Avery, D., Deenick, E. & Hodgkin, P. Intrinsic differences in the proliferation of naive and memory human B cells as a mechanism for enhanced secondary immune responses. J. Immunol. 170, 686–694 (2003).

    Article  CAS  Google Scholar 

  30. Takatsu, K. Interleukin 5 and B cell differentiation. Cytokine Growth Factor Rev. 9, 25–35 (1998).

    Article  CAS  Google Scholar 

  31. Rush, J.S. & Hodgkin, P.D. B cells activated via CD40 and IL-4 undergo a division burst but require continued stimulation to maintain division, survival and differentiation. Eur. J. Immunol. 31, 1150–1159 (2001).

    Article  CAS  Google Scholar 

  32. Hasbold, J. et al. Quantitative analysis of lymphocyte differentiation and proliferation in vitro using carboxyfluorescein diacetate succinimidyl ester. Immunol. Cell. Biol. 77, 516–522 (1999).

    Article  CAS  Google Scholar 

  33. Snapper, C.M., Kehry, M.R., Castle, B.E. & Mond, J.J. Multivalent, but not divalent, antigen receptor cross-linkers synergize with CD40 ligand for induction of Ig synthesis and class switching in normal murine B cells. A redefinition of the TI-2 vs T cell-dependent antigen dichotomy. J. Immunol. 154, 1177–1187 (1995).

    CAS  PubMed  Google Scholar 

  34. Rush, J.S., Hasbold, J. & Hodgkin, P.D. Cross-linking surface Ig delays CD40 ligand- and IL-4-induced B cell Ig class switching and reveals evidence for independent regulation of B cell proliferation and differentiation. J. Immunol. 168, 2676–2682 (2002).

    Article  CAS  Google Scholar 

  35. Hargreaves, D.C. et al. A coordinated change in chemokine responsiveness guides plasma cell movements. J. Exp. Med. 194, 45–56 (2001).

    Article  CAS  Google Scholar 

  36. Shaffer, A.L. et al. Blimp-1 orchestrates plasma cell differentiation by extinguishing the mature B cell gene expression program. Immunity 17, 51–62 (2002).

    Article  CAS  Google Scholar 

  37. Muramatsu, M. et al. Class switch recombination and hypermutation require activation-induced cytidine deaminase (AID), a potential RNA editing enzyme. Cell 102, 553–563 (2000).

    Article  CAS  Google Scholar 

  38. Kelso, A. et al. Heterogeneity in lymphokine profiles of CD4+ and CD8+ T cells and clones activated in vivo and in vitro. Immunol. Rev. 123, 85–114 (1991).

    Article  CAS  Google Scholar 

  39. Kehry, M.R. & Castle, B. Regulation of CD40 ligand expression and use of recombinant CD40 ligand for studying B cell growth and differentiation. Semin. Immunol. 6, 287–294 (1994).

    Article  CAS  Google Scholar 

  40. Lyons, A.B. & Parish, C.R. Determination of lymphocyte division by flow cytometry. J. Immunol. Methods 171, 131–137 (1994).

    Article  CAS  Google Scholar 

  41. Lyons, A.B., Hasbold, J. & Hodgkin, P.D. Flow cytometric analysis of cell division history using dilution of carboxyfluorescein diacetate succinimidyl ester, a stably integrated fluorescent probe. Methods Cell Biol. 63, 375–398 (2001).

    Article  CAS  Google Scholar 

  42. Sedgwick, J.D. & Holt, P.G. The ELISA-plaque assay for the detection and enumeration of antibody- secreting cells. An overview. J. Immunol. Methods 87, 37–44 (1986).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank M. Kehry and B. Castle for gifts of reagents used in this study. Supported by the National Health and Medical Research Council of Australia.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Philip D Hodgkin.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hasbold, J., Corcoran, L., Tarlinton, D. et al. Evidence from the generation of immunoglobulin G–secreting cells that stochastic mechanisms regulate lymphocyte differentiation. Nat Immunol 5, 55–63 (2004). https://doi.org/10.1038/ni1016

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni1016

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing