We hypothesized that tenascin expression is increased in pleural inflammatory and fibrotic diseases and that its expression can be used as a marker of active pleural involvement. For this purpose we analyzed 71 histological samples of inflammatory and fibrotic pleura from patients with asbestos-induced pleural reaction (n = 6), postcardiac injury syndrome (n = 6), parapneumonic infection and/or empyema (n = 23), tuberculosis (n = 5, rheumatoid disease (n = 1), and fibrosis with inflammation of unknown etiology (n = 30). All 71 cases were studied by immunohistochemistry for tenascin. In 19 selected cases tenascin mRNA in situ hybridization was also performed. In every case, tenascin was increased by immunohistochemistry. Most prominent immunoreactivity was detected in areas of newly formed fibrosis. Increased tenascin mRNA expression by in situ hybridization was detected in the individual cells of the newly formed fibrosis underneath the fibrinous exudate. The tenascin mRNA-positive cells localized in areas in which by immunohistochemical studies the cells were positive for α-smooth muscle actin, desmin, and vimentin, suggesting a myofibroblast phenotype. Tenascin mRNA expression was also seen less frequently in areas in which some cells were positive for cytokeratin. These cells might represent mesothelial cells entrapped in the inflammatory lesion. Alternatively, they might represent fibroblast-type cells with aberrant cytokeratin expression. We conclude that in pleural inflammatory and fibrotic diseases tenascin immunoreactivity is increased and tenascin mRNA-positive cells localized mainly in the areas of myofibroblast- and, less often, mesothelial-type cells, suggesting that mainly myofibroblasts and, less commonly, also mesothelial cells might be responsible for tenascin expression in pleural inflammatory and fibrotic diseases.