Temporal stability of local lymph node assay responses to hexyl cinnamic aldehyde

The local lymph node assay is an alternative method for the prospective identification of chemicals that have the potential to cause skin sensitization. Activity in the assay is measured as a function of proliferative responses by draining lymph node cells induced by topical exposure of mice to the test chemical. Positive responses are defined as those where a test chemical, at one or more application concentrations, is able to induce a stimulation index of 3 or greater compared with concurrent vehicle-treated control values. Although the method has been evaluated extensively, the stability over time of responses induced in the local lymph node assay has not previously been addressed formally. It was the purpose of the investigations described here to consider this issue and, to this end, responses provoked in the local lymph node assay by hexyl cinnamic aldehyde (HCA) - a skin sensitizing chemical of moderate potency - were assessed in five separate experiments conducted in a single laboratory over a 10-month period. In each case, HCA elicited a positive response. Although some significant inter-experimental variation was recorded, this was attributable entirely to the stimulation by HCA of slightly more vigorous responses in one of the five experiments. When the results of this experiment were excluded from the data set, significant variations were lost. Finally, for each experiment an EC3 value was derived, this being the estimated concentration of test chemical required to induce a stimulation index of 3. Similar EC3 values were derived in each experiment. These data demonstrate the relative stability over time of activity in the local lymph node assay.