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)

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. © 2017, Isfahan University of Medical Sciences(IUMS). All Rights Reserved.