We have previously shown that one of the co-factors required for generation of T-cell responses, B7.1, is variably expressed on melanoma cells. In the present studies we have examined the expression of another important co-factor in T-cell responses, viz., CD40, and investigated regulation of its expression and possible function(s). PCR analysis revealed mRNA for CD40 in all 18 cell lines established from metastatic melanoma and the majority of those from 6 primary melanoma. CD40 protein was detectable in approximately 50% of the cell lines by flow cytometry and in sections from only 2 of 20 melanoma. Expression of CD40 protein was increased in 2 of 3 cell lines with constitutive CD40 expression by interferon-gamma but not by granulocyte/macrophage colony-stimulating factor, interleukin-2 or tumor necrosis factor-alpha. Interaction of monoclonal antibody with CD40 on melanoma cells resulted in an increase in their cell division but did not increase expression of the costimulatory factor B7. Our results suggest that CD40 expression on melanoma may have important effects on their biology. The influence of CD40 expression on T-cell responses to melanoma remains to be investigated.