Comparison of B-Cells Differentiation to Plasmablasts at Presence of Anti-Human CD40 and Anti-Immunoglobulin M f (ab)'2 or Lipopolysaccharide and Anti-Human CD40 Stimulators (In-Vitro)

Document Type : Original Article (s)

Authors

1 MSc Student, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 PhD Student, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Transformation and differentiation of activated B-cell to plasmacells and also memory cells depend on signaling from B-cell receptors. The signals from antigen and cytokine receptors on the surface of B cells lead to induce the expression of specific transcription factors, which finally determine the fate of B cells.Methods: Peripheral blood mononuclear cells (PBMC) were isolated via ficoll gradient and then purified B cells were separated using magnetic-activated cell sorting (MACS). Pure B cells were cultured in Roswell Park Memorial Institute 1640 (RPMI1640) culture media at the presence of purified anti-human CD40 antibody and anti-immunoglobulin M f (ab)´2 or lipopolysaccharide (LPS) and anti-human CD40 antibody that induced B-cells differentiation to plasmablasts which was assessed with 3 markers (CD38+, CD27+, IgM-) and analyzed via flow cytometry.Findings: In stimulation of B cells with purified anti-human CD40 antibody and anti-IgM f (ab)´2 or LPS through cross-linking B-cell receptor, the majority of B cells remained alive and differentiated to another lineage of B cells (plasmablast: CD38+, CD27+, IgM-). There was no significant statistical difference between expressions of plasmablast markers in two states of stimulation.Conclusion: B cells can be stimulated and differentiated to plasmablasts in vitro similar to in vivo condition. However, to achieve the best outcome in the differentiation of B cells, we should consider the nature of stimulator, the time of incubation, and the type of stimulators.

Keywords

References

  1. Kondo M. Lymphoid and myeloid lineage commitment in multipotent hematopoietic progenitors. Immunol Rev 2010; 238(1): 37-46.
  2. Cooper MD. The early history of B cells. Nat Rev Immunol 2015; 15(3): 191-7.
  3. Schenkein HA, Ruddy S. The role of immunoglobulins in alternative complement pathway activation by zymosan. I. Human IgG with specificity for Zymosan enhances alternative pathway activation by zymosan. J Immunol 1981; 126(1): 7-10.
  4. Janeway Jr CA, Travers P, Walport M, Shlomchik MJ. The humoral immune response. The distribution and functions of immunoglobulin isotypes. In: Janeway Jr CA, Travers P, Walport M, Shlomchik MJ, editors. Immunobiology. 5th ed. New York, NY: Garland Science; 2001.
  5. Manz RA, Lohning M, Cassese G, Thiel A, Radbruch A. Survival of long-lived plasma cells is independent of antigen. Int Immunol 1998; 10(11): 1703-11.
  6. Nutt SL, Hodgkin PD, Tarlinton DM, Corcoran LM. The generation of antibody-secreting plasma cells. Nat Rev Immunol 2015; 15(3): 160-71.
  7. Warnatz K, Schlesier M. Flowcytometric phenotyping of common variable immunodeficiency. Cytometry B Clin Cytom 2008; 74(5): 261-71.
  8. Maiga RI, Bonnaure G, Rochette JT, Neron S. Human CD38hiCD138(+) plasma cells can be generated in vitro from CD40-activated switched-memory B lymphocytes. J Immunol Res 2014; 2014: 635108.
  9. Ribourtout B, Zandecki M. Plasma cell morphology in multiple myeloma and related disorders. Morphologie 2015; 99(325): 38-62.
  10. Harwood NE, Batista FD. Early events in B cell activation. Annu Rev Immunol 2010; 28: 185-210.
  11. Wortis HH, Teutsch M, Higer M, Zheng J, Parker DC. B-cell activation by crosslinking of surface IgM or ligation of CD40 involves alternative signal pathways and results in different B-cell phenotypes. Proc Natl Acad Sci USA 1995; 92(8): 3348-52.
  12. Schilizzi BM, Boonstra R, The TH, de Leij LF. Effect of B-cell receptor engagement on CD40-stimulated B cells. Immunology 1997; 92(3): 346-53.
  13. Hasler P, Zouali M. B cell receptor signaling and autoimmunity. FASEB J 2001; 15(12): 2085-98.
  14. Wiesner M, Zentz C, Mayr C, Wimmer R, Hammerschmidt W, Zeidler R, et al. Conditional immortalization of human B cells by CD40 ligation. PLoS One 2008; 3(1): e1464.
  15. Boeglin E, Smulski CR, Brun S, Milosevic S, Schneider P, Fournel S. Toll-like receptor agonists synergize with CD40L to induce either proliferation or plasma cell differentiation of mouse B cells. PLoS One 2011; 6(10): e25542.
  16. Hua Z, Hou B. TLR signaling in B-cell development and activation. Cell Mol Immunol 2013; 10(2): 103-6.
  17. Xu H, Liew LN, Kuo IC, Huang CH, Goh DL, Chua KY. The modulatory effects of lipopolysaccharide-stimulated B cells on differential T-cell polarization. Immunology 2008; 125(2): 218-28.
  18. Van BK, Herman J, Boon L, Waer M, Sprangers B, Louat T. Comparative In Vitro Immune Stimulation Analysis of Primary Human B Cells and B Cell Lines. J Immunol Res 2016; 2016: 5281823.
  19. Patterson HC, Kraus M, Kim YM, Ploegh H, Rajewsky K. The B cell receptor promotes B cell activation and proliferation through a non-ITAM tyrosine in the Igalpha cytoplasmic domain. Immunity 2006; 25(1): 55-65.
  20. Anbazhagan K, Rabbind SA, Isabelle P, Stella I, Celine AD, Bissac E, et al. Human pre-B cell receptor signal transduction: evidence for distinct roles of PI3kinase and MAP-kinase signalling pathways. Immun Inflamm Dis 2013; 1(1): 26-36.
  21. Nomura J, Inui S, Yamasaki T, Kataoka S, Maeda K, Nakanishi K, et al. Anti-CD40 monoclonal antibody induces the proliferation of murine B cells as a B-cell mitogen through a distinct pathway from receptors for antigens or lipopolysaccharide. Immunol Lett 1995; 45(3): 195-203.
  22. Saito T, Kitayama D, Sakamoto A, Tsuruoka N, Arima M, Hatano M, et al. Effective collaboration between IL-4 and IL-21 on B cell activation. Immunobiology 2008; 213(7): 545-55.
  23. Moser JM, Upton JW, Gray KS, Speck SH. Ex vivo stimulation of B cells latently infected with gammaherpesvirus 68 triggers reactivation from latency. J Virol 2005; 79(8): 5227-31.
Journal of Isfahan Medical School
Volume 35, Issue 427 - Serial Number 427
March and April 2017
Pages 440-446

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History

  • Receive Date: 08 February 2017
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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