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

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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: 25 May 2024
  • Accept Date: 06 April 2022

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