Immunofluorescence and immunoperoxidase methods were used to identify constituents and products of cultured human bronchial epithelial cells and fibroblasts. Epithelial cells, but not fibroblasts, from patients with blood Types A or B reacted to the respective antisera to either the A or B blood group antigens. However, neither the epithelial cells nor the fibroblasts from patients with blood type O[H] reacted with the anti-H antisera. Epithelial cells in primary culture reacted with antibody to prekeratin proteins from human stratum corneum and fibroblasts did not react. Moreover, keratin filaments were assembled in vitro from proteins isolated from the epithelial cells. These immunological and biochemical data support previous morphological observations that human bronchial epithelial cells in primary culture shift progressively from a mucociliary epithelium to a keratinizing epithelium. Epithelial cells and fibroblasts could also be identified by their reactivity to anti-collagen antibodies. Fibroblasts reacted strongly with antibodies to Types I and III collagens and epithelial cells did not. On the other hand, epithelial cells reacted weakly with antibodies to Type IV collagen and fibroblasts were completely negative. Both epithelial cells and fibroblasts reacted with antibody to fibronectin; however, the distribution of fibronectin differed in the two cell types. In epithelial cells, fibronectin was restricted to the cell surface, whereas in fibroblasts it was found on the cell surface and in the extracellular matrix where fibrils of fibronectin were both cell associated and deposited on the surface of the dish where the fibroblasts had migrated.