Interlaboratory validation of the modified murine local lymph node assay based on adenosine triphosphate measurement

Omori, Takashi; Idehara, Kenji; Kojima, Hajime; Sozu, Takashi; Arima, Kazunori; Goto, Hisaharu; Hanada, Tomohiko; Ikarashi, Yoshiaki; Inoda, Taketo; Kanazawa, Yukiko; Kosaka, Toshio; Maki, Eiji; Morimoto, Takashi; Shinoda, Shinsuke; Shinoda, Norihiro; Takeyoshi, Masahiro; Tanaka, Masashi; Uratani, Mamoru; Usami, Masayoshi; Yamanaka, Atsushi; Yoneda, Tomofumi; Yoshimura, Isao; Yuasa, Akihiro

doi:10.1016/j.vascn.2008.05.001

Cited by 13 publications

(8 citation statements)

References 34 publications

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“…Non-radioactive endpoints, such as lymph node weight, cell proliferation and ear swelling, have been described by several authors (Ehling et al 2005a, b;Suda et al 2002;for review: Anderson et al 2011). Two versions of the LLNA with non-radioactive endpoints, the LLNA: DA and the BrdU-ELISA-version, are currently validated and available as draft OECD guidelines (Omori et al 2008;Kojima et al 2011). Ulrich et al (2001 described a two-tiered murine local lymph node assay and proposed that it is useful to discriminate between contact allergenic potential and skin irritation potential.…”

Section: Discussionmentioning

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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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: Discussionmentioning

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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“…After thorough evaluation and reviews, these tests were adopted by The Organisation for Economic Co-operation and Development (OECD) as stand-alone skin sensitizing tests: Test Guideline (TG) 429, TG442A, and TG442B (Kimber et al, 1986;Basketter et al, 1996;Gerberich et al, 2000;Dean et al, 2001;Haneke et al, 2001;Takeyoshi et al, 2001;Sailstad et al, 2001;OECD TG No. 429, 2002, 2010bYamashita et al, 2005;Idehara et al, 2008;Omori et al, 2008;ICCVAM, 2010;OECD TG No. 442A, 2010a;OECD TG No.…”

Section: Introductionmentioning

confidence: 99%

Further development of LLNA:DAE method as stand-alone skin-sensitization testing method and applied for evaluation of relative skin-sensitizing potency between chemicals

Yamashita

Shinoda²,

Hagiwara³

et al. 2015

J. Toxicol. Sci.

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-To date, there has been no well-established local lymph node assay (LLNA) that includes an elicitation phase. Therefore, we developed a modified local lymph node assay with an elicitation phase (LLNA:DAE) to discriminate true skin sensitizers from chemicals that gave borderline positive results and previously reported this assay. To develop the LLNA:DAE method as a useful stand-alone testing method, we investigated the complete procedure for the LLNA:DAE method using hexyl cinnamic aldehyde (HCA), isoeugenol, and 2,4-dinitrochlorobenzene (DNCB) as test compounds. We defined the LLNA:DAE procedure as follows: in the dose-finding test, four concentrations of chemical applied to dorsum of the right ear on days 1, 2, and 3 and dorsum of both ears on day 10. Ear thickness and skin irritation score were measured on days 1, 3, 5, 10, and 12. Local lymph nodes were excised and weighed on day 12. The test dose for the primary LLNA:DAE study was selected as the dose that gave the highest left ear lymph node weight in the dose-finding study, or the lowest dose that produced a left ear lymph node of over 4 mg. This procedure was validated using nine different chemicals. Furthermore, qualitative relationship was observed between the degree of elicitation response in the left ear lymph node and the skin sensitizing potency of 32 chemicals tested in this study and the previous study. These results indicated that LLNA:DAE method was as first LLNA method that was able to evaluate the skin sensitizing potential and potency in elicitation response.

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“…The LLNA:DA employs ATP measurement as an index of lymph node cell proliferation, with principles of evaluation similar to those of the LLNA. The reliability and relevance of the LLNA:DA were examined by a committee and have been validated by 17 laboratories belonging to that committee (Omori et al, 2008). These LLNA methods have several advantages over the GPMT/BA, including generation of quantitative data, a reduction in test time, a reduction in the required number of animals, and an improvement in animal welfare (Haneke et al, 2001;Basketter et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Development of LLNA:DAE: a new local lymph node assay that includes the elicitation phase, discriminates borderline-positive chemicals, and is useful for cross-sensitization testing

Yamashita

Shinoda²,

Hagiwara³

et al. 2014

J. Toxicol. Sci.

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-We developed a new local lymph node assay (LLNA) that includes the elicitation phase termed LLNA:DAE for discrimination of borderline-positive chemicals as classified by the LLNA modified by Daicel based on ATP content (LLNA:DA) and for cross-sensitization testing. Although the LLNA:DA method could help identify skin sensitizers, some skin irritants classified as non-sensitizers by the LLNA were classified as borderline positive. In addition, the evaluation for the cross-sensitization potential between chemicals was impossible. In the LLNA:DAE procedure, test group of mice received four applications of chemicals on the dorsum of the right ear for induction and one application on the dorsum of the left ear for elicitation. Control group of mice received one chemical application on the dorsum of the left ear. We evaluated the sensitizing potential by comparing the weights of the lymph nodes from the left ears between the two groups. The results of using the LLNA:DAE method to examine 24 chemicals, which contained borderline-positive chemicals, were consistent with those from the LLNA method, except for nickel chloride (NiCl 2 ). Two chemical pairs, 2,4-dinitrochlorobenzene (DNCB) with 2,4-dinitrofluorobenzene (DNFB) and hydroquinone (HQ) with p-benzoquinone (p-BQ), showed clear cross-sensitization with each other, while another chemical pair, DNFB with hexylcinnamic aldehyde (HCA) did not. Taken together, our results suggest that the LLNA:DAE method is useful for discriminating borderline-positive chemicals and for determining chemical cross-sensitization.

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Interlaboratory validation of the modified murine local lymph node assay based on adenosine triphosphate measurement

Cited by 13 publications

References 34 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)

Further development of LLNA:DAE method as stand-alone skin-sensitization testing method and applied for evaluation of relative skin-sensitizing potency between chemicals

Development of LLNA:DAE: a new local lymph node assay that includes the elicitation phase, discriminates borderline-positive chemicals, and is useful for cross-sensitization testing

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