Staphylococcus δ-toxin induces allergic skin disease by activating mast cells

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Abstract

Atopic dermatitis is a chronic inflammatory skin disease that affects 15–30% of children and approximately 5% of adults in industrialized countries1. Although the pathogenesis of atopic dermatitis is not fully understood, the disease is mediated by an abnormal immunoglobulin-E immune response in the setting of skin barrier dysfunction2. Mast cells contribute to immunoglobulin-E-mediated allergic disorders including atopic dermatitis3. Upon activation, mast cells release their membrane-bound cytosolic granules leading to the release of several molecules that are important in the pathogenesis of atopic dermatitis and host defence4. More than 90% of patients with atopic dermatitis are colonized with Staphylococcus aureus in the lesional skin whereas most healthy individuals do not harbour the pathogen5. Several staphylococcal exotoxins can act as superantigens and/or antigens in models of atopic dermatitis6. However, the role of these staphylococcal exotoxins in disease pathogenesis remains unclear. Here we report that culture supernatants of S. aureus contain potent mast-cell degranulation activity. Biochemical analysis identified δ-toxin as the mast cell degranulation-inducing factor produced by S. aureus. Mast cell degranulation induced by δ-toxin depended on phosphoinositide 3-kinase and calcium (Ca2+) influx; however, unlike that mediated by immunoglobulin-E crosslinking, it did not require the spleen tyrosine kinase. In addition, immunoglobulin-E enhanced δ-toxin-induced mast cell degranulation in the absence of antigen. Furthermore, S. aureus isolates recovered from patients with atopic dermatitis produced large amounts of δ-toxin. Skin colonization with S. aureus, but not a mutant deficient in δ-toxin, promoted immunoglobulin-E and interleukin-4 production, as well as inflammatory skin disease. Furthermore, enhancement of immunoglobulin-E production and dermatitis by δ-toxin was abrogated in KitW-sh/W-sh mast-cell-deficient mice and restored by mast cell reconstitution. These studies identify δ-toxin as a potent inducer of mast cell degranulation and suggest a mechanistic link between S. aureus colonization and allergic skin disease.

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Figure 1: S. aureus δ-toxin induces MC degranulation in vitro and in vivo.
Figure 2: MC degranulation induced by δ-toxin depends on Ca2+ influx/phosphoinositide 3-kinase pathway, but is independent of Syk.
Figure 3: Antigen-independent IgE signalling enhances δ-toxin-induced MC activation.
Figure 4: Staphyloccocus δ-toxin promotes IgE production and inflammatory skin disease by mast cells.

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Acknowledgements

We thank S. Koonse for animal husbandry, J. Whitfield for enzyme-linked immunosorbent assays, S. Meshinchi for electron microscopy, V. Basrur for mass spectrometry, K. Kidwell for advice with statistical analysis, M. K. Oyoshi and R. S. Geha for experimental advice, V. Y. Tan for help with constructing the LAC P3-lux strain and A. Burberry for reviewing the manuscript. Y.N. was supported by fellowships from the Chiba University Global COE Program, the Cell Science Research Foundation and the Kanae Foundation for the Promotion of Medical Science. J.O. and K.B.C. were supported by Department of Veterans Affairs Merit Award I01BX000429. A.E.V., G.Y.C.C. and M.O. were supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH). This work supported by NIH grants R01AR059688 to G.N. and R01HL062996 to J.B.T. and funds to the Michigan Comprehensive Cancer Center Immunology Monitoring Core from the University of Michigan’s Cancer Center Support Grant.

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Y.N., N.I. and G.N. designed the research. Y.N. conducted the experiments and analysed data with the help of R.M.-P., S.M.C. and M.H. J.O., K.B.C., J.B.T. and M.J.M. generated and provided critical reagents or material. A.E.V, G.Y.C.C. and M.O. engineered bacterial strains. Y.N. and G.N. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Gabriel Núñez.

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Nakamura, Y., Oscherwitz, J., Cease, K. et al. Staphylococcus δ-toxin induces allergic skin disease by activating mast cells. Nature 503, 397–401 (2013). https://doi.org/10.1038/nature12655

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