Skip to main content

Advertisement

SpringerLink
Log in

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

  • Immunotoxicology
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

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)

References

  • Aeby P, Ashikaga T, Bessou-Touya S, Schepky A, Gerberick F, Kern P, Marrec-Fairley M, Maxwell G, Ovigne JM, Sakaguchi H, Reisinger K, Tailhardat M, Martinozzi-Teissier S, Winkler P (2010) Identifying and characterizing chemical skin sensitizers without animal testing: Colipa’s research and method development program. Toxicol In Vitro 24:1465–1473

    Article  PubMed  CAS  Google Scholar 

  • Ahuja V, Schreiber C, Platzek T, Stahlmann R (2009a) Investigation of the sensitising and cross-sensitising potential of textile dyes and beta-lactam antibiotics using a biphasic mice local lymph node assay. Arch Toxicol 83:691–699

    Article  PubMed  CAS  Google Scholar 

  • Ahuja V, Wanner R, Platzek T, Stahlmann R (2009b) Appraisal of the sensitising potential of orally and dermally administered mercaptobenzothiazole by a biphasic protocol of the local lymph node assay. Arch Toxicol 83:933–939

    Article  PubMed  CAS  Google Scholar 

  • Ahuja V, Platzek T, Fink H, Sonnenburg A, Stahlmann R (2010) Study of the sensitising potential of various textile dyes using a biphasic murine local lymph node assay. Arch Toxicol 84:709–718

    Article  PubMed  CAS  Google Scholar 

  • American Apparel & Footwear Association (2011) Restricted substances list (RSL). (Release 8), http://www.apparelandfootwear.org/Resources/RestrictedSubstances.asp

  • Anderson SE, Siegel PD, Meade BJ (2011) The LLNA: a brief review of recent advances and limitations. J Allergy (Cairo) 2011:424203

    Google Scholar 

  • Basketter DA, Kan-King-Yu D, Dierkes P, Jowsey IR (2007) Does irritation potency contribute to the skin sensitization potency of contact allergens? Cutan Ocul Toxicol 26:279–286

    Article  PubMed  CAS  Google Scholar 

  • Bordel-Gomez MT, Miranda-Romero A, Castrodeza-Sanz J (2010) Epidemiology of contact dermatitis: prevalence of sensitization to different allergens and associated factors. Actas Dermosifiliogr 101:59–75

    Article  PubMed  Google Scholar 

  • Ehling G, Hecht M, Heusener A, Huesler J, Gamer AO, van Loveren H, Maurer T, Riecke K, Ulrich P, Vandebriel R, Vohr HW (2005a) An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: 2nd round. Toxicology 212:69–79

    Article  PubMed  CAS  Google Scholar 

  • Ehling G, Hecht M, Heusener A, Huesler J, Gamer AO, van Loveren H, Maurer T, Riecke K, Ullmann L, Ulrich P, Vandebriel R, Vohr HW (2005b) An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: first round. Toxicology 212:60–68

    Article  PubMed  CAS  Google Scholar 

  • Federal Institute for Risk Assessment (2007) Introduction to the problems surrounding garment textiles. BfR Information (18), http://www.bfr.bund.de/cm/349/introduction_to_the_problems_surrounding_garment_textiles.pdf

  • Hatch KL (1984) Chemicals and textiles. Part I: dermatological problems related to fibre content and dyes (review). Text Res J 54:664–682

    Article  Google Scholar 

  • Hatch KL, Motschi H, Maibach HI (2003) Disperse dyes in fabrics of patients patch-test-positive to disperse dyes. Am J Contact Dermat 14:205–212

    PubMed  Google Scholar 

  • Hausen BM (1993) Contact allergy to disperse blue 106 and blue 124 in black “velvet” clothes. Contact Dermat 28:169–173

    Article  PubMed  CAS  Google Scholar 

  • Hausen BM, Menezes BF (1986) Disperse blue 106, a strong sensitizer. Contact Dermat 15:102–103

    Article  PubMed  CAS  Google Scholar 

  • Hausen BM, Sawall EM (1989) Sensitization experiments with textile dyes in guinea pigs. Contact Dermat 20:27–31

    Article  PubMed  CAS  Google Scholar 

  • Kojima H, Takeyoshi M, Sozu T, Awogi T, Arima K, Idehara K, Ikarashi Y, Kanazawa Y, Maki E, Omori T, Yuasa A, Yoshimura I (2011) Inter-laboratory validation of the modified murine local lymph node assay based on 5-bromo-2′-deoxyuridine incorporation. J Appl Toxicol 31:63–74

    Article  PubMed  CAS  Google Scholar 

  • Marks JG Jr (1986) Occupational skin disease in hairdressers. Occup Med 1:273–284

    PubMed  CAS  Google Scholar 

  • McFadden JP, Basketter DA (2000) Contact allergy, irritancy and ‘danger’. Contact Dermat 42:123–127

    Article  CAS  Google Scholar 

  • Omori T, Idehara K, Kojima H, Sozu T, Arima K, Goto H, Hanada T, Ikarashi Y, Inoda T, Kanazawa Y, Kosaka T, Maki E, Morimoto T, Shinoda S, Shinoda N, Takeyoshi M, Tanaka M, Uratani M, Usami M, Yamanaka A, Yoneda T, Yoshimura I, Yuasa A (2008) Interlaboratory validation of the modified murine local lymph node assay based on adenosine triphosphate measurement. J Pharmacol Toxicol Methods 58:11–26

    Article  PubMed  CAS  Google Scholar 

  • Python F, Goebel C, Aeby P (2007) Assessment of the U937 cell line for the detection of contact allergens. Toxicol Appl Pharmacol 220:113–124

    Article  PubMed  CAS  Google Scholar 

  • Sadhra S, Duhra P, Foulds IS (1989) Occupational dermatitis from Synacril Red 3b liquid (CI Basic Red 22). Contact Dermat 21:316–320

    Article  CAS  Google Scholar 

  • Sailstad DM (2002) Murine local lymph node assay: an alternative test method for skin hypersensitivity testing. Lab Anim (NY) 31:36–41

    Google Scholar 

  • Sakaguchi H, Ashikaga T, Miyazawa M, Yoshida Y, Ito Y, Yoneyama K, Hirota M, Itagaki H, Toyoda H, Suzuki H (2006) Development of an in vitro skin sensitization test using human cell lines; human Cell Line Activation Test (h-CLAT). II. An inter-laboratory study of the h-CLAT. Toxicol In Vitro 20:774–784

    Article  PubMed  CAS  Google Scholar 

  • Schreiner M, Peiser M, Briechle D, Stahlmann R, Zuberbier T, Wanner R (2007) A loose-fit coculture of activated keratinocytes and dendritic cell-related cells for prediction of sensitizing potential. Allergy 62:1419–1428

    Article  PubMed  CAS  Google Scholar 

  • Schreiner M, Peiser M, Briechle D, Stahlmann R, Zuberbier T, Wanner R (2008) A new dendritic cell type suitable as sentinel of contact allergens. Toxicology 249:146–152

    Article  PubMed  CAS  Google Scholar 

  • Seidenari S, Giusti F, Massone F, Mantovani L (2002) Sensitization to disperse dyes in a patch test population over a five-year period. Am J Contact Dermat 13:101–107

    Article  PubMed  CAS  Google Scholar 

  • Stahlmann R, Wegner M, Riecke K, Kruse M, Platzek T (2006) Sensitising potential of four textile dyes and some of their metabolites in a modified local lymph node assay. Toxicology 219:113–123

    Article  PubMed  CAS  Google Scholar 

  • Suda A, Yamashita M, Tabei M, Taguchi K, Vohr HW, Tsutsui N, Suzuki R, Kikuchi K, Sakaguchi K, Mochizuki K, Nakamura K (2002) Local lymph node assay with non-radioisotope alternative endpoints. J Toxicol Sci 27:205–218

    Article  PubMed  CAS  Google Scholar 

  • Ulrich P, Streich J, Suter W (2001) Intralaboratory validation of alternative endpoints in the murine local lymph node assay for the identification of contact allergic potential: primary ear skin irritation and ear-draining lymph node hyperplasia induced by topical chemicals. Arch Toxicol 74:733–744

    Article  PubMed  CAS  Google Scholar 

  • Wanner R, Sonnenburg A, Quatchadze M, Schreiner M, Peiser M, Zuberbier T, Stahlmann R (2010) Classification of sensitizing and irritative potential in a combined in vitro assay. Toxicol Appl Pharmacol 245:211–218

    Article  PubMed  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  1. Varun Ahuja

    Present address: Pre-clinical Safety Assessment (Novel Drug Discovery & Development), Lupin Ltd, Pune, India

Authors and Affiliations

  1. Institute of Clinical Pharmacology and Toxicology, Charité—Universitätsmedizin Berlin, Luisenstr. 7, 10117, Berlin, Germany

    Anna Sonnenburg, Varun Ahuja & Ralf Stahlmann

  2. Department of Internal Medicine, Bundeswehr Hospital, Berlin, Germany

    Maximilian Schreiner

  3. Federal Institute for Risk Assessment (BfR), Max-Dohrn-Str. 8-10, 10589, Berlin, Germany

    Thomas Platzek

Authors
  1. Anna Sonnenburg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Varun Ahuja
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maximilian Schreiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thomas Platzek
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ralf Stahlmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ralf Stahlmann.

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-012-0811-9

Keywords

Search

Navigation