Recent reports suggest that production of interleukin-12 (IL-12) by dendritic cells and keratinocytes may play an important part in contact hypersensitivity reactions. In the present study we investigated mRNA and protein expression of IL-12 in human skin lymph derived from normal untreated skin (n = 5) and from the induction phase of allergic contact dermatitis (CD) (n = 5). mRNA levels were determined at various time points in the lymph cells by a nested reverse transcriptase-polymerase chain reaction method. Time course analysis reproducibly revealed a constitutive expression of both IL-12 p40 and p35 mRNA in the migrating lymph cells in all volunteers. However, no enhancement of the IL-12 mRNA signal was found during the induction phase of allergic CD. Furthermore, as determined by a sensitive ELISA technique, IL-12 protein was not detectable in 60 lymph samples derived from normal untreated skin or in 68 lymph samples obtained during the induction phase of allergic CD at any time point of the lymph cannulation. In conclusion, our findings indicate that no significant protein levels of IL-12 are washed out from the skin into the afferent lymph or are produced and released by migrating lymph cells during the induction phase of allergic CD in vivo.