The cytological diagnosis of malignant lymphoma can be extremely difficult because the cytological features of the malignant cells in small cell and mixed small and large cell lymphomas may be indistinguishable from those of reactive lymphoid cells. We have examined the usefulness of the avidin biotin immunoperoxidase technique and a battery of antibodies to T and B cell markers to the diagnosis of lymphoma in cytological specimens. We conclude that immunocytochemistry is very useful in the cytological diagnosis of non-Hodgkin's lymphoma. Further, it is possible to diagnose the vast majority of lymphomas using only the immunoglobulin light chain markers kappa and lambda and the T-cell markers CD5, CD3, CD4 and CD8. We are extending the utilization of lymphoid markers to fine needle aspiration specimens of lymph nodes. Fine needle aspiration may obviate the need for repeat biopsies in patients with recurrent lymphomas. Another project utilizing immunocytochemistry as an adjunct to routine light microscopic cytologic diagnosis, involves distinguishing polyoma viral effects from atypia secondary to cyclophosphamide therapy in urine specimens. A large population of patients followed at the NIH are receiving cyclophosphamide therapy on an on-going basis for the treatment of both benign and malignant disease. Cytomorphologic abnormalities have been described int he urine of cyclophosphamide-treated patients and have been confused cytologically with urinary tract neoplasia, the incidence of which is also increased following cyclophosphamide therapy. Furthermore, the cytologic features of polyoma virus cytopathic effect in the urine also overlap the features of cyclophosphamide effect and neoplasia. We have used immunocytochemistry with a polyclonal antibody to polyoma virus to document the presence of virus in the urine specimens of some patients in order to better define the distinguishing characteristics of cyclophosphamide effect, neoplasia and polyoma virus.