Atopic dermatitis and skin disease
Tight junction defects in patients with atopic dermatitis

https://doi.org/10.1016/j.jaci.2010.10.018Get rights and content

Background

Atopic dermatitis (AD) is characterized by dry skin and a hyperactive immune response to allergens, 2 cardinal features that are caused in part by epidermal barrier defects. Tight junctions (TJs) reside immediately below the stratum corneum and regulate the selective permeability of the paracellular pathway.

Objective

We evaluated the expression/function of the TJ protein claudin-1 in epithelium from AD and nonatopic subjects and screened 2 American populations for single nucleotide polymorphisms in the claudin-1 gene (CLDN1).

Methods

Expression profiles of nonlesional epithelium from patients with extrinsic AD, nonatopic subjects, and patients with psoriasis were generated using Illumina's BeadChips. Dysregulated intercellular proteins were validated by means of tissue staining and quantitative PCR. Bioelectric properties of epithelium were measured in Ussing chambers. Functional relevance of claudin-1 was assessed by using a knockdown approach in primary human keratinocytes. Twenty-seven haplotype-tagging SNPs in CLDN1 were screened in 2 independent populations with AD.

Results

We observed strikingly reduced expression of the TJ proteins claudin-1 and claudin-23 only in patients with AD, which were validated at the mRNA and protein levels. Claudin-1 expression inversely correlated with TH2 biomarkers. We observed a remarkable impairment of the bioelectric barrier function in AD epidermis. In vitro we confirmed that silencing claudin-1 expression in human keratinocytes diminishes TJ function while enhancing keratinocyte proliferation. Finally, CLDN1 haplotype-tagging SNPs revealed associations with AD in 2 North American populations.

Conclusion

Collectively, these data suggest that an impairment in tight junctions contributes to the barrier dysfunction and immune dysregulation observed in AD subjects and that this may be mediated in part by reductions in claudin-1.

Section snippets

Study participants: Expression profiling and validation experiments

The diagnosis of AD was made using the US consensus conference criteria.36 All patients with AD had extrinsic disease, as defined by a serum total IgE level 2 SDs or more of age-dependent norms and a positive multiallergen RAST result (ImmunoCap Phadiatop). Nonatopic healthy subjects were defined as having no personal or family history of atopic diseases, no personal history of chronic skin or systemic diseases, and a serum total IgE level that was 2 SDs or less of age-dependent norms and a

Claudin-1 expression is markedly reduced in nonlesional AD epidermis

To characterize and quantify the expression of human epidermal proteins important for barrier function, we performed gene expression profiling of nonlesional or clinically unaffected epidermis by using blister roofs from patients with AD, patients with psoriasis, and nonatopic subjects (Fig 2, B-D). The Illumina Sentrix HumanRef-8 Chip contained 43 TJ genes (see Table E2 in this article's Online Repository at www.jacionline.org), 8 gap junction genes, and 41 epidermal differentiation complex

Discussion

This is the first report, to our knowledge, implicating a TJ defect in patients with AD, a human skin disease that affects up to 15 million Americans. We demonstrated reduced expression of epidermal claudin-1 in AD nonlesional epidermis (Figs 2 and 3). This was specific for AD and not observed in patients with psoriasis, a TH17-driven inflammatory skin disorder (Fig 2). Although previous psoriasis publications have suggested that TJs might be altered in lesional epidermis, this has not been

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    Supported by the Atopic Dermatitis and Vaccinia Network NIH/NIAID (contract N01 AI40029 and N01 AI40033), the National Eczema Association (A.D., L.A.B.), and the Mary Beryl Patch Turnbull Scholar Program (K.C.B.).

    Disclosure of potential conflict of interest: A. De Benedetto has received research support from the National Eczema Association and the National Institutes of Health (NIH)/National Institute of Allergy and Infectious Diseases (NIAID). N. M. Rafaels has received research support from the NIH/NIAID. C. Cheadle has received research support from the NIH/NIAID. T. Yoshida has received research support from the NIH/NIAID. M. Boguniewicz has received research support from the NIH/NIAID. J. M. Hata has received research support from the NIH/NIAID. L. C. Schneider has received research support from Astellas and Novartis. J. M. Hanifin has received consulting fees from Johnson & Johnson and Otsuka Pharmaceutical Co and has received research support from Astellas Pharma US, Basilea, Novartis, Shionogi USA, the NIH/NIAID, and Taisho. R. L. Gallo has received research support from the NIH/NIAID. N. Novak has received research support from the German Research Council and the NIH/NIAID. T. H. Beaty has received research support from the NIH/NIAID. D. Y. M. Leung has received research support from the NIH/NIAID. K. C. Barnes has received research support from the NIH and Sanofi-Aventis. L. A. Beck has consulted for Regeneron, has received research support from the NIH/NIAID and the National Eczema Association. The rest of the authors have declared that they have no conflict of interest.

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