TLR9 signaling failure renders Peyer's patch regulatory B cells unresponsive to stimulation with CpG oligodeoxynucleotides

Abstract

Intestinal Peyer's patch (PP) regulatory CD21+ B cells (Bregs) suppress TLR9-induced innate immune responses. However, it is not known whether TLR9 activation is regulated in PP Bregs. Here, we investigated the responses of PP Bregs to stimulation with the TLR9 agonist CpG oligodeoxynucleotides (ODN). We observed that PP CD21+ Bregs express high levels of TLR9 mRNA, but fail to proliferate when stimulated with CpG ODN. Furthermore, unlike CD21+ B cells from blood, PP CD21+ Bregs do not secrete IgM or IL-12 following CpG ODN stimulation. We hypothesized that the unresponsiveness of PP Bregs to CpG stimulation was due to an inability of the TLR9 agonist to activate the TLR9 signaling pathway in these cells. This was confirmed by kinome analysis which demonstrated dynamic patterns of phosphorylation of key TLR adaptor proteins such as IRAK1, TAK1, IKK and NF-κB-p65 in CpG-stimulated blood CD21+ B cells, consistent with activation of the TLR9 pathway. In contrast, stimulation of PP CD21+ B regs with CpG ODN resulted in phosphorylation patterns of these adaptor proteins suggestive of inactivation of the TLR9 pathway. The absence of apparent TLR9 signaling events immediately following stimulation indicated that signaling is blocked close to the receptor. Our observations suggest a novel mechanism by which the host regulates TLR responses in TLR-expressing cells with regulatory functions. © 2010 S. Karger AG, Basel.