Molecular mechanisms by which interferon regulatory factor 5 (IRF5) transduces signaling downstream of BCR and BCR/TLR9 pathways

Abstract The transcription factor IRF5 modulates human B cell activation, proliferation, and plasmablast differentiation via an intrinsic mechanism. Targeted knockdown of IRF5 in human primary naïve B cells revealed a significant reduction in CD19+CD86+ B cells, which demarcate early activated B cells, after BCR/TLR9 stimulation, resulting in reduced clonal expansion and plasmablast differentiation. The mechanism(s) by which IRF5 modulates early B cell activation is not known. We previously found that BCR stimulation alone was insufficient to induce IRF5 nuclear translocation (activation), while BCR/TLR9 stimulation induced robust IRF5 activation. Thus, the focus of this study is to determine the mechanism(s) by which IRF5 functions downstream of BCR and dual BCR/TLR9 signaling at the early time points of activation (5 and 15 min). Experiments were performed in human B cells and B cells from wild-type and Irf5−/− mice. Signaling and protein interactions were examined by Western blot, immunoprecipitation, flow cytometry, and biochemical assay. We identified Bruton’s tyrosine kinase (Btk) as a novel interacting partner of IRF5 in both unstimulated and stimulated human and murine B cells. Of note, active phosphorylated Btk was bound to IRF5 only in stimulated cells. Preliminary data also suggest a defect downstream of Btk activation in Irf5−/− B cells. Together, these data indicate a conserved role for IRF5 in the early stages of B cell activation via BCR/TLR9 signaling.