TNF overproduction impairs epithelial staphylococcal response in hyper IgE syndrome
Ian A. Myles, … , Steven M. Holland, Sandip K. Datta
Ian A. Myles, … , Steven M. Holland, Sandip K. Datta
Published July 23, 2018
Citation Information: J Clin Invest. 2018;128(8):3595-3604. https://doi.org/10.1172/JCI121486.
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TNF overproduction impairs epithelial staphylococcal response in hyper IgE syndrome

Abstract

Autosomal dominant hyper IgE syndrome (AD-HIES), or Job’s syndrome, is a primary immune deficiency caused by dominant-negative mutations in STAT3. Recurrent Staphylococcus aureus skin abscesses are a defining feature of this syndrome. A widely held hypothesis that defects in peripheral Th17 differentiation confer this susceptibility has never been directly evaluated. To assess the cutaneous immune response in AD-HIES, we induced suction blisters in healthy volunteers (HVs) and patients with AD-HIES and then challenged the wound with lethally irradiated bacteria. We show that cutaneous production of IL-17A and IL-17F was normal in patients with AD-HIES. Overproduction of TNF-α differentiated the responses in AD-HIES from HVs. This was associated with reduced IL-10 family signaling in blister-infiltrating cells and defective epithelial cell function. Mouse models of AD-HIES recapitulated these aberrant epithelial responses to S. aureus and involved defective epithelial-to-mesenchymal transition (EMT) rather than a failure of bacterial killing. Defective responses in mouse models of AD-HIES and primary keratinocyte cultures from patients with AD-HIES could be reversed by TNF-α blockade and by drugs with reported modulatory effects on EMT. Our results identify these as potential therapeutic approaches in patients with AD-HIES suffering S. aureus infections.

Authors

Ian A. Myles, Erik D. Anderson, Noah J. Earland, Kol A. Zarember, Inka Sastalla, Kelli W. Williams, Portia Gough, Ian N. Moore, Sundar Ganesan, Cedar J. Fowler, Arian Laurence, Mary Garofalo, Douglas B. Kuhns, Mark D. Kieh, Arhum Saleem, Pamela A. Welch, Dirk A. Darnell, John I. Gallin, Alexandra F. Freeman, Steven M. Holland, Sandip K. Datta

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Figure 3

Skin cell cultures from patients with AD-HIES display TNF-α–sensitive defects in wound healing.

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Skin cell cultures from patients with AD-HIES display TNF-α–sensitive de...
(A) 3D organotypic cultures of KCs and FBs from HVs and patients with AD-HIES. (B) IF images (original magnification, ×20) for DAPI (blue) and vimentin (red) and (C) mean intensity of the IF signal of the leading edge of the scratch assay monolayer culture of dermal FBs from HVs and AD-HIES patients. (D) IF images of organotypic cultures, as in A, stained for β-catenin (red), vimentin (green), and DAPI (blue). FBs and KCs were sourced from both HVs and AD-HIES patients, as indicated. Scale bars: 40 μm. (E) Western blot of nuclear and cytoplasmic fractions from KCs obtained from HVs and AD-HIES patients (KCs were cultured with TNF-α or diluent). (F) Band intensity for β-catenin, as in E, normalized to either a histone 3 (nuclear) or β-actin (cytoplasmic) loading control. (G) Area of coverage after 18 to 22 hours of incubation in a scratch assay for KCs (see Supplemental Video 1) and FBs (see Supplemental Video 2). Data are representative of 3 or more independent experiments and displayed as the mean ± SD. n = 3 cell lines per group, indicated by individual dots. Each experiment used at least 2 different cell lines assayed in duplicate. *P < 0.05, **P < 0.01, and ****P < 0.0001, by ANOVA with Bonferroni’s correction, compared with diluent treatment (F), or as indicated. Dil., diluent.