Abstract
Sensitization requires exposure to an allergen with subsequent production of a "danger "signal. In the skin, keratinocytes are the main producers of these signals. To compare dose- and time-effects of cobalt on the viability of and cytokine release from HaCaT cells cultured at low or high calcium. To model two separate states of differentiation of keratinocytes, HaCaT cells were cultured under low or high calcium conditions. HaCaT were exposed to different concentrations of cobalt chloride (10 μm to 5 mM) over time (30 minutes- 48 hours). Cell viability was measured with the Cell-Titer Blue Viability assay. Cytokine production was measured using a bead-based immunoassay and flow cytometry. Gene expression was quantified using qPCR. Data was analyzed by ANOVA and linear mixed model. Viability of the cells was dose- and time-dependent. A linear mixed statistical model showed that cobalt exposure induces increase in IL-6, CXCL8 and CCL2 production over time and whereas increase of IL-6 and a decrease of CCL2 was associated with increasing cobalt chloride concentrations. When comparing the cells incubated under high and low calcium conditions, the more differentiated cells in the high concentration were found to exert a stronger response in terms of IL-6 release. Our data suggest that cobalt chloride triggered an alarm system in HaCaT cells, and proinflammatory cytokines/chemokines were secreted in a dose- and time-dependent manner. When high and low calcium incubations were compared, the difference was seen only for IL-6. These findings indicate that the effect of cobalt chloride on cell toxicity occurs throughout the living epidermis.
Highlights
The prevalence of contact allergy to cobalt among patch tested patients in Sweden was shown to be 4,5% in 2009 [1]
A linear mixed statistical model showed that cobalt exposure induces increase in IL-6, CXCL8 and CCL2 production over time and whereas increase of IL-6 and a decrease of CCL2 was associated with increasing cobalt chloride concentrations
Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production in HaCaT exposed to cobalt
Summary
The prevalence of contact allergy to cobalt among patch tested patients in Sweden was shown to be 4,5% in 2009 [1]. Keratinocytes have the ability to respond to stimuli from our environment and exert inflammatory mediation in the skin through the production of several pro-inflammatory cytokines as a response to various danger signals [10, 11]. These soluble immune-modulating factors from keratinocytes have been shown to be involved in delayedtype hypersensitivity reactions i.e., allergic contact dermatitis making keratinocytes an important component in the initiation of these maladies [12]. Sensitization requires exposure to an allergen with subsequent production of a “danger “signal. In the skin, keratinocytes are the main producers of these signals
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