Abstract
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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Abbreviations
- AAA:
-
p-Aminoacetanilide (CAS No. 122-80-5)
- 7-AAD:
-
7-Aminoactinomycine D (CAS No. 7240-37-1)
- ANT:
-
2-Amino-5-nitrothiazole (CAS No. 121-66-4)
- ApC:
-
2-Amino-p-cresol (CAS No. 95-84-1)
- DB1:
-
Disperse Blue 1 (CAS No. 2475-45-8)
- DB106:
-
Disperse Blue 106 (CAS No. 12223-01-7)
- DB124:
-
Disperse Blue 124 (CAS No. 61951-51-7)
- DB35:
-
Disperse Blue 35 (CAS No. 12222-75-2)
- DCs:
-
Dendritic cells
- DCh:
-
2,6-Dichloro-4-nitroaniline (CAS No. 99-30-9)
- DCrc:
-
Dendritic cell-related cells
- DMSO:
-
Dimethylsulfoxide
- DO3:
-
Disperse Orange 3 (CAS No. 730-40-5)
- DO37:
-
Disperse Orange 37/76 (CAS No. 13301-61-6)
- DR1:
-
Disperse Red 1 (CAS No. 2872-52-8)
- DY3:
-
Disperse Yellow 3 (CAS No. 2832-40-8)
- EAE:
-
2-(N-ethylanilino)-ethanol (CAS No. 95-50-2)
- EC50:
-
Substance concentration causing a half-maximal increase of CD86 expression
- FACS:
-
Fluorescence-activated cell sorting
- FCS:
-
Foetal calf serum
- GMPT:
-
Guinea pig maximization test
- KCs:
-
Keratinocytes
- KGM-2:
-
Keratinocyte growth medium 2
- LCSA:
-
Loose-fit coculture-based sensitization assay
- LLNA:
-
Murine local lymph node assay
- MFI:
-
Mean fluorescence intensity
- PBMCs:
-
Peripheral blood mononuclear cells
- PE:
-
Phycoerythrin
- pNA:
-
p-Nitroaniline (CAS No. 100-01-6)
- pPD:
-
1,4-Phenylenediamine (CAS No. 106-50-3)
- TNBS:
-
2,4,6-Trinitrobenzenesulfonic acid (CAS No. 2508-19-2)
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Acknowledgments
In memoriam. The in vitro assay LCSA (loose-fit coculture-based sensitization assay) is a result of the research activities of Priv.-Doz. Dr. rer. nat. Reinhard Wanner, who passed away unexpectedly at the age of 51 on 2nd April 2010. Since 2008, Dr. Wanner had been the coordinator of the Master Degree Program in Toxicology at the Charité—Universitätsmedizin Berlin. His scientific activities focussed on the immunologic aspects of the skin. Over the last few years, he and his co-workers published a series of papers on LCSA experiments and described the usefulness of this in vitro test, which has great potential to be used as an alternative to the LLNA or other animal experiments. Data presented in this manuscript further underline the robustness and usefulness of the test; they derive from experiments that he had initiated in early 2010. We feel deeply obliged to our colleague, friend and mentor, and we will carry on his ideas into the future. Studies were supported by a grant from the Federal Institute for Risk Assessment (Bundesinstitut für Riskobewertung, BfR), Berlin, Germany. We are also grateful for a grant of the Sonnenfeld Stiftung, Berlin, which financed the FACS Calibur.
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Sonnenburg, A., Ahuja, V., Schreiner, M. et al. Assessment of the sensitizing potential of textile disperse dyes and some of their metabolites by the loose-fit coculture-based sensitization assay (LCSA). Arch Toxicol 86, 733–740 (2012). https://doi.org/10.1007/s00204-012-0811-9
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DOI: https://doi.org/10.1007/s00204-012-0811-9