In this paper, the effect of Si-Ni-San, a famous traditional Chinese prescription, on ear contact sensitivity was examined. Si-Ni-San significantly inhibited the ear swelling when administered during the induction phase of picryl-chloride-induced ear contact sensitivity in mice. The adhesion to type I collagen of isolated spleen cells was significantly decreased in the Si-Ni-San group, especially in the presence of protein kinase C activator, phorbol 12,13-dibutyrate. The inhibition of adhesion was seen in purified T cells from the spleen, as well as in Jurkat cells. Furthermore, the adhesion to collagen involves the production of matrix metalloproteinase-2 and -9 in spleen cells, and the oral administration of Si-Ni-San remarkably reduced the matrix metalloproteinase production. Three drug-pairs composed in Si-Ni-San were used for comparison. The combination of Chaihu and Shaoyao showed a similar effect to Si-Ni-San, while Chaihu and Zhishi, and Shaoyao and Gancao only showed a tendency in most situations. These results suggest that Si-Ni-San may contribute to the treatment of immunologically related diseases by down-regulating the activation and function of T lymphocytes, especially the cell adhesion and matrix metalloproteinase production. Its effect is mainly displayed by the combination of Chaihu and Shaoyao.