Human lymphocytes from a lymph node draining the tumor-bearing area of a patient with a large primary squamous cell carcinoma of the oral mucosa were fused with the nonproducer mouse myeloma, NS-1, to produce interspecies hybridomas. Of 95 hybridoma culture supernatants tested, 23 contained from 0.5 to 50 micrograms/ml of human IgM or IgG. Six supernatant fluids containing greater than 15 micrograms/ml of Ig were tested by indirect immunoperoxidase and immunofluorescence against sections of the autologous carcinoma. Five IgM (lambda) monoclonal antibodies stained the cytoplasm of autologous and allogeneic squamous carcinoma cells. All five monoclonal antibodies stained all layers of normal epidermis but each antibody stained the superficial keratin layer most intensely. Two of the five hybridoma antibodies were further tested. Both antibodies stained all types of normal epithelium; a network of fibers characteristic of intermediate filaments in cultured squamous carcinoma cells and cultured fibroblasts; Z lines in skeletal muscle; and axons in peripheral nerve fibers. We conclude that all five IgM monoclonal antibodies recognize cytokeratins associated with the autologous squamous cell carcinoma. Two of the five hybridoma antibodies recognize an antigenic determinant common to all types of intermediate filament proteins. These data indicate that cytokeratins released by squamous carcinoma cells induced an antibody response in this patient.