Classification of sensitizing and irritative potential in a combined in-vitro assay

Wanner, R.; Sonnenburg, Anna; Quatchadze, Maria; Schreiner, Maximilian; Peiser, Matthias; Zuberbier, Torsten; Stahlmann, Ralf

doi:10.1016/j.taap.2010.02.019

Cited by 23 publications

(16 citation statements)

References 27 publications

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“…Those with an EC50% value from 50 to 1,000 M were considered to be weakly irritative. Substances that did not reach the 50% threshold or with EC50% values above 1,000 M were rated as non-irritative (Wanner et al 2010).…”

Section: Categorizationmentioning

confidence: 99%

“…We have recently developed an in vitro assay based on a coculture of primary human keratinocytes and allogenic dendritic cell-related cells (DCrc), which is capable of identifying the sensitizing and irritative potential of substancesthe loose-Wt coculture-based sensitization assay (LCSA) (Schreiner et al 2007;Wanner et al 2010). Using the LCSA, we have tested eight textile dyes, which are listed in the latest RSL (AAfA 2011) and which the Federal Institute for Risk Assessment recommends not to use in textiles (BfR 2007).…”

Section: Introductionmentioning

confidence: 99%

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Assessment of the sensitizing potential of textile disperse dyes and some of their metabolites by the loose-fit coculture-based sensitization assay (LCSA)

et al. 2012

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Certain textile disperse dyes are known to cause allergic reactions of the human skin. Here, we examined 8 disperse dyes and 7 products of azo-cleavage of these dyes in an in vitro assay. We used the loose-fit coculture-based sensitization assay (LCSA) of primary human keratinocytes and of allogenic dendritic cell-related cells for combined testing of the sensitizing and irritative properties of these substances. The obtained data were compared to data generated in a modified version of the local lymph node assay by our working group. Disperse Blue 1 (DB1), p-nitroaniline (pNA) and p-aminoacetanilide (AAA) showed no sensitizing potential under our experimental conditions. Disperse Blue 124 (DB124), Disperse Yellow 3 (DY3), Disperse Orange 37/76 (DO37), Disperse Blue 106 (DB106), Disperse Red 1 (DR1), 2-amino-p-cresol (ApC), Disperse Orange 3 (DO3) and 2,6-dichloro-4-nitroaniline (DCh) were categorized as extreme sensitizers. Para-phenylenediamine (pPD) was categorized as strong sensitizer, and 2-amino-5-nitrothiazole (ANT) and 2-(N-ethylanilino)-ethanol (EAE) as weak sensitizers. All dyes, except for DB1, and ApC turned out to be strong irritants. DB1, ANT and DCh showed only weak irritative potential. PPD, pNA, EAE and AAA did not show any irritative effect at the concentration range tested. These results correlate with data derived from the modified version of LLNA and human data. Therefore, the LCSA represents a suitable test system to simultaneously analyse two crucial properties of substances relevant for allergy induction.

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Section: Categorizationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Assessment of the sensitizing potential of textile disperse dyes and some of their metabolites by the loose-fit coculture-based sensitization assay (LCSA)

et al. 2012

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“…This assay is unique among those involving DC differentiation in that the DC are generated from monocytes within a growing layer of primary keratinocytes [67,68]. A panel of chemicals have already been tested and the LCSA was able to quantify the irritative and sensitizing potential within 5 days [69]. A comparison of half-maximal increases in mean fluorescence intensity for CD86 demonstrated concordance between the LCSA and LLNA for substances such as DNCB and PPD, which are categorized as extreme and strong, respectively.…”

Section: Topic 1: Chemical-reactive T Cellsmentioning

confidence: 92%

T-cell recognition of chemicals, protein allergens and drugs: towards the development of in vitro assays

Martin

Esser

Schmucker

et al. 2010

Cell. Mol. Life Sci.

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131

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Chemicals can elicit T-cell-mediated diseases such as allergic contact dermatitis and adverse drug reactions. Therefore, testing of chemicals, drugs and protein allergens for hazard identification and risk assessment is essential in regulatory toxicology. The seventh amendment of the EU Cosmetics Directive now prohibits the testing of cosmetic ingredients in mice, guinea pigs and other animal species to assess their sensitizing potential. In addition, the EU Chemicals Directive REACh requires the retesting of more than 30,000 chemicals for different toxicological endpoints, including sensitization, requiring vast numbers of animals. Therefore, alternative methods are urgently needed to eventually replace animal testing. Here, we summarize the outcome of an expert meeting in Rome on 7 November 2009 on the development of T-cell-based in vitro assays as tools in immunotoxicology to identify hazardous chemicals and drugs. In addition, we provide an This article is dedicated to the memory of our colleague and friend Reinhard Wanner, died 2 April 2010. Cellular and Molecular Life Sciences overview of the development of the field over the last two decades.

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“…Categories for irritants are: "irritative" (<50 µmol/l), "weakly irritative" (<1000 µmol/l), "not irritative" (>1000 µmol/l). EC values were calculated based on concentration curves of test substances [for details, see Wanner et al (2010)]. …”

Section: Facs Analysismentioning

confidence: 99%

“…Application of sensitizers to this coculture leads to up-regulation of CD86 on dendritic cells. Earlier studies have shown that the LCSA already proved the capacity to discriminate between sensitizers and irritants (Wanner et al 2010), the latter often tested false positive in the LLNA. Metal allergens, often unrecognized using standard LLNA protocols, are also detected by our in vitro assay (Schreiner et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Unsaturated compounds induce up-regulation of CD86 on dendritic cells in the in vitro sensitization assay LCSA

et al. 2015

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Unsaturated compounds are known to cause false-positive reactions in the local lymph node assay (LLNA) but not in the guinea pig maximization test. We have tested a panel of substances (succinic acid, undecylenic acid, 1-octyn-3-ol, fumaric acid, maleic acid, linoleic acid, oleic acid, alpha-linolenic acid, squalene, and arachidonic acid) in the loose-fit coculture-based sensitization assay (LCSA) to evaluate whether unspecific activation of dendritic cells is a confounder for sensitization testing in vitro. Eight out of 10 tested substances caused significant up-regulation of CD86 on dendritic cells cocultured with keratinocytes and would have been classified as sensitizers; only succinic acid was tested negative, and squalene had to be excluded from data analysis due to poor solubility in cell culture medium. Based on human data, only undecylenic acid can be considered a true sensitizer. The true sensitizing potential of 1-octyn-3-ol is uncertain. Fumaric acid and its isomer maleic acid are not known as sensitizers, but their esters are contact allergens. A group of 18- to 20-carbon chain unsaturated fatty acids (linoleic acid, oleic acid, alpha-linolenic acid, and arachidonic acid) elicited the strongest reaction in vitro. This is possibly due to the formation of pro-inflammatory lipid mediators in the cell culture causing nonspecific activation of dendritic cells. In conclusion, both the LLNA and the LCSA seem to provide false-positive results for unsaturated fatty acids. The inclusion of T cells in dendritic cell-based in vitro sensitization assays may help to eliminate false-positive results due to nonspecific dendritic cell activation. This would lead to more accurate prediction of sensitizers, which is paramount for consumer health protection and occupational safety.

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Classification of sensitizing and irritative potential in a combined in-vitro assay

Cited by 23 publications

References 27 publications

Assessment of the sensitizing potential of textile disperse dyes and some of their metabolites by the loose-fit coculture-based sensitization assay (LCSA)

Assessment of the sensitizing potential of textile disperse dyes and some of their metabolites by the loose-fit coculture-based sensitization assay (LCSA)

T-cell recognition of chemicals, protein allergens and drugs: towards the development of in vitro assays

Unsaturated compounds induce up-regulation of CD86 on dendritic cells in the in vitro sensitization assay LCSA

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