Cutaneous hypersensitivity reactions can develop against antigens delivered through the epidermis (contact dermatitis) or through the blood vessels (e.g., drug eruptions). On routine histology alone, it is not always possible to determine the route of the antigen. Langerhans cells (LC) are the main antigen-presenting cells in contact dermatitis. Dermal dendrocytes (DC) are antigen-presenting cells and may be involved in dermal reactions. We tested the hypothesis that there is a difference between dermatitis due to external and internal antigen sources with regard to the number or function of LC and DC. In 85 cases of dermatitis, numbers of S100 and HLA-DR reactive cells per linear millimetre of epidermis were counted. The amount of epidermal spongiosis was evaluated qualitatively. In 35 cases, the number of DC per mm2 (as defined by Factor XIIIa expression) was evaluated. The patients were then divided into two groups based on whether the final clinical evaluation considered the dermatitis to be secondary to an external (35 cases) or internal antigen (50 cases). Dermatitis due to external antigens had significantly more LC/mm and more frequent HLA-DR expression than dermatitis due to internal antigens, mean +/- SEM; 21.2+/-2.04 vs. 9.1+/-1.02 (p<0.00001) and 16.3+/-2.49 vs. 6.0+/-0.92 (p=0.0001), respectively. Spongiosis was more marked in external antigen cases. DC were more numerous in internal than in external antigen cases, but the differences were not statistically significant. In our model, determination of numbers of LC/mm is the variable with the highest power to discriminate between internal and internal sources. Quantification of HLA-DR+ LC and degree of spongiosis provide little additional discriminatory power.