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Activation of c-Kit in dendritic cells regulates T helper cell differentiation and allergic asthma

Abstract

Dendritic cells (DCs) are integral to the differentiation of T helper cells into T helper type 1 TH1, TH2 and TH17 subsets. Interleukin-6 (IL-6) plays an important part in regulating these three arms of the immune response by limiting the TH1 response and promoting the TH2 and TH17 responses. In this study, we investigated pathways in DCs that promote IL-6 production. We show that the allergen house dust mite (HDM) or the mucosal adjuvant cholera toxin promotes cell surface expression of c-Kit and its ligand, stem cell factor (SCF), on DCs. This dual upregulation of c-Kit and SCF results in sustained signaling downstream of c-Kit, promoting IL-6 secretion. Intranasal administration of antigen into c-Kit–mutant mice or neutralization of IL-6 in cultures established from the lung-draining lymph nodes of immunized wild-type mice blunted the TH2 and TH17 responses. DCs lacking functional c-Kit or those unable to express membrane-bound SCF secreted lower amounts of IL-6 in response to HDM or cholera toxin. DCs expressing nonfunctional c-Kit were unable to induce a robust TH2 or TH17 response and elicited diminished allergic airway inflammation when adoptively transferred into mice. Expression of the Notch ligand Jagged-2, which has been associated with TH2 differentiation, was blunted in DCs from c-Kit–mutant mice. c-Kit upregulation was specifically induced by TH2- and TH17-skewing stimuli, as the TH1-inducing adjuvant, CpG oligodeoxynucleotide, did not promote either c-Kit or Jagged-2 expression. DCs generated from mice expressing a catalytically inactive form of the p110δ subunit of phosphatidylinositol-3 (PI3) kinase (p110D910A) secreted lower amounts of IL-6 upon stimulation with cholera toxin. Collectively, these results highlight the importance of the c-Kit–PI3 kinase–IL-6 signaling axis in DCs in regulating T cell responses.

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Figure 1: Cholera toxin and HDM stimulate IL-6 secretion from DCs in vitro and in vivo.
Figure 2: Increased expression of c-Kit in CT - and HDM-stimulated BMDCs.
Figure 3: c-Kit+ but not c-Kit BMDCs resulting from CT stimulation skew T cells predominantly toward a TH2 and TH17 response.
Figure 4: CT- or HDM-induced c-Kit signaling regulates IL-6 production and the expression of Notch ligands.
Figure 5: Impaired TH2 and TH17 response in vivo after BMDCs containing mutant c-Kit, and consequently secreting less IL-6, were adoptively transferred intratracheally (IT) into WT mice.
Figure 6: Decreased expression of soluble SCF, and increased expression of the membrane-bound form, in CT-stimulated DCs.

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Acknowledgements

We dedicate this study to the memory of Professor Ray Wu, who inspired us to tread the untrodden path. We would like to thank K. Murphy (Washington University, St. Louis) for DO11.10 transgenic mice, A. Henry and B. Dixon-McCarthy for technical assistance with the adoptive transfer experiment and assessment of airway inflammation, M. Zheng and J.K. Kolls for help with the Klebsiella infection model, S. Plevy for providing us with breeding pairs of p110δD910A/D910A mice, and S. Shapiro for critically reading the manuscript. This work was supported by US National Institutes of Health grants HL 060207 and HL 069810 (to P.R.), HL 077430 and AI 048927 (to A.R.) and HL 084932 (to P.R. and A.R.).

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N.K., T.B.O. and M.F. performed in vitro and in vivo experiments. M.P. and M.Y. performed microarray and PCR experiments. B.V. provided PI3 kinase–mutant mice and critically read the manuscript. N.K. and T.B.O. also participated in study design and N.K. prepared the manuscript. P.R. and A.R. conceived and designed the study, analyzed the data and wrote the manuscript.

Corresponding authors

Correspondence to Anuradha Ray or Prabir Ray.

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Krishnamoorthy, N., Oriss, T., Paglia, M. et al. Activation of c-Kit in dendritic cells regulates T helper cell differentiation and allergic asthma. Nat Med 14, 565–573 (2008). https://doi.org/10.1038/nm1766

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