TLR2 deficiency promotes IgE and inhibits IgG1 class-switching following ovalbumin sensitization

Background To explore the roles of Toll-like receptor (TLR)2 in Th2 cytokine production and immunoglobulin (Ig) class switching following ovalbumin (OVA) sensitization. Methods TLR2(-/-) and wild-type C57BL/6 mice were sensitized by intraperitoneal injection with OVA. Lung pathology was assessed by hematoxylin and eosin staining. Abundance of interleukin (IL)4, IL5, IL13, and IL21 transcripts in the lungs was quantified by RT-PCR. OVA-specific IgG1, IgG2a, IgG2b, IgE and IgM were quantified by enzyme-linked immunosorbent assay. Phosphorylated signal transducer and activator of transcription (STAT)3 in lung tissue was detected by immunohistochemistry staining and nuclear factor (NF) kappa B activation was measured by immunofluorescence staining. STAT3 activation was inhibited using cryptotanshinone (CPT) treatment. Germline transcripts (I mu-C mu, I gamma-C gamma, I alpha-C alpha or I epsilon-C epsilon), post-recombination transcripts (I mu-C gamma, I mu-C alpha or I mu- C epsilon) and mature transcripts (V(H)DJ(H)-C gamma, V(H)DJ(H)-C alpha or V(H)DJ(H)-C epsilon) were analyzed from splenic B cells of OVA-sensitized wild-type mice (with or without CPT treatment) and TLR2(-/-) mice (with or without IL21 treatment). Results The lungs of TLR2(-/-) mice showed a lesser degree of inflammation than wild-type mice after OVA sensitization. Following OVA sensitization, levels of IL4, IL13, and IL21, but not IL5, were significantly lower in TLR2(-/-) compared with wild-type mice. Moreover, OVA-specific IgG1 and IgE titers were markedly lower and higher, respectively, in TLR2(-/-) mice. TLR2 deficiency inhibited STAT3 activation but not NF-kappa B p65 activation. CPT treatment reduced IgG1 titers via inhibition of Stat3 phosphorylation. Both TLR2 knockout and CPT treatment reduced the frequencies of I gamma 1-C gamma 1, I gamma 3-C gamma 3 and I alpha-C alpha transcripts, but IL21 treatment compensated for the effects of TLR2 deficiency. Conclusion These results suggest a role of TLR2 in restricting OVA-sensitized lung inflammation via promotion of IgG1 and inhibition of IgE class switching regulated by IL21 and STAT3.

Automated summary

The study found that TLR2 plays a role in limiting lung inflammation post-OVA sensitization by promoting IgG1 and inhibiting IgE class switching, with IL21 and STAT3 regulating this process.