KC and macrophage-inflammatory protein-2 (MIP-2) are CXC chemokines that exhibit distinct temporal patterns of expression in the skin following surgical injury. In situ hybridization analysis demonstrates that these two chemokines are expressed by distinct cell types at different times following injury. Dermal fibroblasts and endothelial cells are primarily responsible for KC expression in the skin 6 h following surgery. In contrast, MIP-2 production appears to be restricted to infiltrating inflammatory leukocytes including neutrophils and monocytes, which appear later in the response. This cell type-specific pattern of chemokine expression is recapitulated in vitro using isolated primary- and long-term-cultured cell types. Primary dermal fibroblasts stimulated with interleukin-1alpha express predominantly KC and very little MIP-2, and peritoneal exudate neutrophils produce as much or more MIP-2 as KC following stimulation in vitro. Although a collection of exogenous stimuli can induce expression of KC and MIP-2, the quantitative ratio for expression reflects the cell type and not the stimulus. The selective expression of KC over MIP-2 in endothelial cells results from markedly greater KC gene transcription and not from alterations in the rate of mRNA decay. These results demonstrate that distinct CXC chemokines show restricted expression in myeloid versus nonmyeloid cell types and that patterns of chemokine expression at sites of inflammation in vivo reflect the temporally ordered contribution of these distinct cell types.