Journal of Isfahan Medical School

Journal of Isfahan Medical School

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
Sanaz Afshar-Qasemloo 1
Nafiseh Esmaeil 2
Mazdak Ganjalikhani-Hakemi 2
Abbas Rezaei 3
Reza Yazdani 4
Faezeh Abbasi-Rad 1
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
B-Cells
Plasmablast
B-cell differentiation factor receptor
Lipopolysaccharides
Flow cytometry
  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

  • Receive Date 08 February 2017
  • Accept Date 06 April 2022