Previous investigations have indicated that different classes of chemical allergen provoke discrete immune responses in mice characterized by the development of selective cytokine secretion patterns by draining lymph node cells. It was found that, in mice exposed chronically to chemical respiratory allergens, lymph node cells produced high levels of mitogen-inducible interleukin 4 (IL-4), a cytokine necessary for the development of IgE antibody responses. In contrast, exposure of mice to contact allergens resulted in only low levels of IL-4 production. The studies described here were conducted to determine whether differentially induced expression of IL-4 messenger RNA was independent of the need for mitogen stimulation, unlike secretion of IL-4 protein. Mice were exposed to concentrations of trimellitic anhydride (TMA), a respiratory allergen, or to 2,4-dinitrochlorobenzene (DNCB), a contact allergen, under conditions of equivalent immunogenicity and where the former, but not the latter, resulted in significant production of mitogen-inducible IL-4 protein. The steady state levels of IL-4 mRNA in draining lymph nodes were measured by reverse transcription-polymerase chain reaction after a single or repeated daily application of the test chemical to the ears of chronically sensitized mice. Expression of mRNA for IL-4 was evaluated relative to the expression of a housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase. Both chemicals elicited increased steady state levels of IL-4 mRNA by draining lymph node cells compared with vehicle-treated and naive controls. However, using two different treatment protocols, a chronic and an abbreviated chronic protocol and at all times examined, the elevation in IL-4 mRNA steady state levels induced by TMA was greater than that observed with DNCB. These data provide further support for the differential stimulation by contact and respiratory chemical allergens of cytokine secretion patterns in mice and demonstrate that the divergent production of IL-4 in response to these classes of allergen is at least partly transcriptionally regulated.