5. OECD Test No. 442A: Skin sensitization. Local lymph node stimulation test (DA)

The local lymph node stimulation assay: DA (ELGL: DA) is a non-radioactive variant of the ELGL method that aims to identify sensitizing chemicals and measure the lymphocyte proliferation they induce in the auricular lymph nodes. This method, described in mice (CBA/J strain), is based on the bioluminescence quantification of adenosine triphosphate (ATP) content, an indicator of this proliferation.

A minimum of four animals are used per dose group, with a minimum of three concentrations of the test substance, plus a negative control group treated only with the vehicle, and a positive control if necessary. The experimental protocol lasts eight days. The ATP content must be quantified within 30 minutes of euthanasia. The procedure applied, from lymph node excision to ATP measurement, must be identical for each animal and must not exceed 20 minutes.

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