MUTZ-3 derived Langerhans cells in human skin equivalents show differential migration and phenotypic plasticity after allergen or irritant exposure.

After allergen or irritant exposure, Langerhans cells (LC) undergo phenotypic changes and exit the epidermis. In this study we describe the unique ability of MUTZ-3 derived Langerhans cells (MUTZ-LC) to display similar phenotypic plasticity as their primary counterparts when incorporated into a physiologically relevant full-thickness skin equivalent model (SE-LC). We describe differences and similarities in the mechanisms regulating LC migration and plasticity upon allergen or irritant exposure. The skin equivalent consisted of a reconstructed epidermis containing primary differentiated keratinocytes and CD1a(+) MUTZ-LC on a primary fibroblast-populated dermis. Skin equivalents were exposed to a panel of allergens and irritants. Topical exposure to sub-toxic concentrations of allergens (nickel sulfate, resorcinol, cinnamaldehyde) and irritants (Triton X-100, SDS, Tween 80) resulted in LC migration out of the epidermis and into the dermis. Neutralizing antibody to CXCL12 blocked allergen-induced migration, whereas anti-CCL5 blocked irritant-induced migration. In contrast to allergen exposure, irritant exposure resulted in cells within the dermis becoming CD1a(-)/CD14(+)/CD68(+) which is characteristic of a phenotypic switch of MUTZ-LC to a macrophage-like cell in the dermis. This phenotypic switch was blocked with anti-IL-10. Mechanisms previously identified as being involved in LC activation and migration in native human skin could thus be reproduced in the in vitro constructed skin equivalent model containing functional LC. This model therefore provides a unique and relevant research tool to study human LC biology in situ under controlled in vitro conditions, and will provide a powerful tool for hazard identification, testing novel therapeutics and identifying new drug targets.

[1]  D. Ingber,et al.  Reconstituting Organ-Level Lung Functions on a Chip , 2010, Science.

[2]  S. H. van der Burg,et al.  Intradermal Delivery of TLR Agonists in a Human Explant Skin Model: Preferential Activation of Migratory Dendritic Cells by Polyribosinic-Polyribocytidylic Acid and Peptidoglycans , 2013, The Journal of Immunology.

[3]  T. D. de Gruijl,et al.  MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34+ precursors. , 2002, Blood.

[4]  T. D. de Gruijl,et al.  High susceptibility of c-KIT+CD34+ precursors to prolonged doxorubicin exposure interferes with Langerhans cell differentiation in a human cell line model , 2011, Cancer Immunology, Immunotherapy.

[5]  F. Nestle,et al.  Resident CD141 (BDCA3)+ dendritic cells in human skin produce IL-10 and induce regulatory T cells that suppress skin inflammation , 2012, The Journal of experimental medicine.

[6]  M Régnier,et al.  Reactivity of Langerhans cells in human reconstructed epidermis to known allergens and UV radiation. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[7]  P. Das,et al.  Skin irritants and contact sensitizers induce Langerhans cell migration and maturation at irritant concentration , 2006, Experimental dermatology.

[8]  Emanuela Corsini,et al.  An epidermal equivalent assay for identification and ranking potency of contact sensitizers. , 2013, Toxicology and applied pharmacology.

[9]  T. Takezawa,et al.  Reconstruction of three-dimensional human skin model composed of dendritic cells, keratinocytes and fibroblasts utilizing a handy scaffold of collagen vitrigel membrane. , 2009, Toxicology in vitro : an international journal published in association with BIBRA.

[10]  S. Gibbs,et al.  Autocrine regulation of re-epithelialization after wounding by chemokine receptors CCR1, CCR10, CXCR1, CXCR2, and CXCR3. , 2012, The Journal of investigative dermatology.

[11]  O. Damour,et al.  Evolutive skin reconstructions: from the dermal collagen-glycosaminoglycan-chitosane substrate to an immunocompetent reconstructed skin. , 2006, Bio-medical materials and engineering.

[12]  S. Spiekstra,et al.  Induction of cytokine (interleukin‐1α and tumor necrosis factor‐α) and chemokine (CCL20, CCL27, and CXCL8) alarm signals after allergen and irritant exposure , 2005, Experimental dermatology.

[13]  T. D. de Gruijl,et al.  IL-10 Conditioning of Human Skin Affects the Distribution of Migratory Dendritic Cell Subsets and Functional T Cell Differentiation , 2013, PloS one.

[14]  A. Mikhailova,et al.  Immunostimulatory conventional dendritic cells evolve into regulatory macrophage-like cells. , 2012, Blood.

[15]  S. Spiekstra,et al.  CCL5 and CCL20 mediate immigration of Langerhans cells into the epidermis of full thickness human skin equivalents. , 2012, European journal of cell biology.

[16]  Susan Gibbs,et al.  Technical Advance: Langerhans cells derived from a human cell line in a full‐thickness skin equivalent undergo allergen‐induced maturation and migration , 2011, Journal of leukocyte biology.

[17]  D. Schmitt,et al.  Integration of Langerhans cells into a pigmented reconstructed human epidermis. , 1997, The Journal of investigative dermatology.

[18]  S. Meijer,et al.  Characterization of four conventional dendritic cell subsets in human skin-draining lymph nodes in relation to T-cell activation. , 2011, Blood.

[19]  Eric Perrier,et al.  Effects of solar ultraviolet radiation on engineered human skin equivalent containing both Langerhans cells and dermal dendritic cells. , 2007, Tissue engineering.

[20]  E. Klechevsky,et al.  The differential production of cytokines by human Langerhans cells and dermal CD14(+) DCs controls CTL priming. , 2012, Blood.

[21]  Erwin L Roggen,et al.  In vitro approaches for detection of chemical sensitization. , 2014, Basic & clinical pharmacology & toxicology.

[22]  T. D. de Gruijl,et al.  CXCL12 is essential for migration of activated Langerhans cells from epidermis to dermis , 2008, European journal of immunology.

[23]  Thomas Vogl,et al.  Crucial role for human Toll-like receptor 4 in the development of contact allergy to nickel , 2010, Nature Immunology.

[24]  C. Borrebaeck,et al.  Functional and transcriptional profiling of MUTZ‐3, a myeloid cell line acting as a model for dendritic cells , 2006, Immunology.

[25]  T. D. de Gruijl,et al.  Epidermis‐to‐dermis migration of immature Langerhans cells upon topical irritant exposure is dependent on CCL2 and CCL5 , 2010, European journal of immunology.

[26]  H. Pinedo,et al.  A CD34+ human cell line model of myeloid dendritic cell differentiation: evidence for a CD14+CD11b+ Langerhans cell precursor , 2006, Journal of leukocyte biology.

[27]  S. Santegoets,et al.  Human dendritic cell line models for DC differentiation and clinical DC vaccination studies , 2008, Journal of leukocyte biology.

[28]  Lisa M. Ebert,et al.  Cutaneous CXCL14 targets blood precursors to epidermal niches for Langerhans cell differentiation. , 2005, Immunity.

[29]  Yong‐jun Liu,et al.  Mouse and human dendritic cell subtypes , 2002, Nature Reviews Immunology.

[30]  D. Schmitt,et al.  Phenotypic and functional outcome of human monocytes or monocyte-derived dendritic cells in a dermal equivalent. , 2001, The Journal of investigative dermatology.

[31]  N. Fusenig,et al.  Mutual induction of growth factor gene expression by epidermal-dermal cell interaction , 1993, The Journal of cell biology.

[32]  R. Zaino,et al.  Changes in Lymphocyte and Langerhans Cell Populations in Allergic and Irritant Contact Dermatitis , 1987, International journal of dermatology.

[33]  T. D. de Gruijl,et al.  Irritant-induced migration of Langerhans cells coincides with an IL-10-dependent switch to a macrophage-like phenotype. , 2011, The Journal of investigative dermatology.

[34]  Valérie Zuang,et al.  Alternative (non-animal) methods for cosmetics testing: current status and future prospects—2010 , 2011, Archives of Toxicology.

[35]  T. D. de Gruijl,et al.  A Postmigrational Switch among Skin-Derived Dendritic Cells to a Macrophage-Like Phenotype Is Predetermined by the Intracutaneous Cytokine Balance1 , 2006, The Journal of Immunology.

[36]  S. Gibbs,et al.  Progress on the development of human in vitro dendritic cell based assays for assessment of the sensitizing potential of a compound. , 2009, Toxicology and applied pharmacology.

[37]  S. Spiekstra,et al.  Comparison of a novel CXCL12/CCL5 dependent migration assay with CXCL8 secretion and CD86 expression for distinguishing sensitizers from non-sensitizers using MUTZ-3 Langerhans cells. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.