Alternatives to Animal Testing and Experimentation
Online ISSN : 2185-4726
Print ISSN : 1344-0411
ISSN-L : 1344-0411
Original Article
Development of a Modified Local Lymph Node Assay using ATP Measurement as an Endpoint
Kunihiko YamashitaKenji IdeharaNorio FukudaGaku YamagishiNaoki Kawada
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2005 年 11 巻 2 号 p. 136-144

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Several proposed modified local lymph node assays (LLNAs) use a non-RI method, but such methods tend to yield lower stimulation indices (SIs) than those obtained with the standard LLNA when used to assess moderate sensitizers. To address this issue, we propose a modified LLNA in which mice are exposed to the test material four times, instead of three times, and adenosine triphosphate (ATP) content is used as a measure of proliferation of lymph node cells (LNCs). The chemicals tested were 2.4-dinitro-chlorobenzene (DNCB), eugenol, and a-hexyl cinnamic aldehyde (HCA), which are classified as strong-to-moderate sensitizers in the LLNA. Methyl salicylate (MS) was used as an example of a non-sensitizer. Using an SI of 3 as a cut-off, as recommended in the standard LLNA, DNCB, eugenol and HCA were classified as positive, while MS was classified as negative using the ATP-based non-RI method. Furthermore, there were no marked differences in SI levels for 10% eugenol and 10% HCA (a moderate sensitizer) between the non-RI method and the standard LLNA. Thus, it is reasonable to con-clude that the non-RI method proposed in the present study could be a valid alternative to the standard LLNA.

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© 2005 Japanese Society for Alternative to Animal Experiments
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