Analysis of interleukin-1α (IL-1α) and interleukin-8 (IL-8) expression and release in in vitro reconstructed human epidermis for the prediction of in vivo skin irritation and/or sensitization

• Published in: Toxicology in vitro Vol. 17; no. 3; pp. 311 - 321
• Main Authors: Coquette, A., Berna, N., Vandenbosch, A., Rosdy, M., De Wever, B., Poumay, Y.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.06.2003; Elsevier Science
• Subjects: Allergic diseases; Biological and medical sciences; Chemical and industrial products toxicology. Toxic occupational diseases; Immunopathology; Medical sciences; Metals and various inorganic compounds; Skin allergic diseases. Stinging insect allergies; Toxicology; Various organic compounds; BC; NiSO 4; DMSO; DNCB; MTT; PDC; RHE; LLNA; GM-CSF; RT-PCR; SLS; PBS; DNFB; IL-15; IL-8; IL-12; IP-10; TNBS; IL-1α; IL-6; IL-10; IL-7; TGF; TNF-α; OXA; PCR; BA; Benzoic acid; Human; Nickel Sulfates; Cell culture; Toxicity; Cytokine; Prediction; Interleukin 1α; Benzalkonium chloride; Cytotoxicity; Epidermis; Gene expression; In vitro; Dose activity relation; Keratinocyte; Skin; Interleukin 8; Release; Allergen; Irritation
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/S0887-2333(03)00019-5

In the present study, reconstructed human epidermis (RHE) was used as an in vitro model to discriminate 1-chloro-2,4-dinitrobenzene (DNCB), nickel sulfate (NiSO 4), oxazolone (OXA), 2,4-dinitrofluorobenzene (DNFB) and 2,4,6-trinitrobenzenesulfonic acid (TNBS) as skin sensitizers from benzalkonium chloride (BC), benzoic acid (BA) and sodium lauryl sulfate (SLS) as skin irritants. Our criteria were (a) the differential IL-1α and IL-8 synthesis and release (b) cytotoxicity assessment by MTT assay. When the RHE are topically treated with the sensitizers, very low levels of extra- and intracellular IL-1α are observed although they induce significant cytotoxicity. In contrast, they exhibit a sharp maximum of IL-8 release. In the presence of the tested irritants, we observe the inverse cytokine release profile, although they induce dose-dependent cytotoxicity profiles similar to those observed with the sensitizers. Finally, IL-1α mRNA upregulation is observed after topical application of both sensitizers and irritants, but only the latter significantly increase extracellular IL-1α. In conclusion, our results suggest that the associated determination of IL-8, with IL-1α, and MTT conversion are at least necessary to discriminate and classify, in a single assay, irritant and sensitizing agents and represent a potential in vitro alternative to two classical in vivo assays.