Comparisonbetween the Cellular Proteinsof HairyCell Leukemiaand the LeukemicPhaseof Other LymphoproliferativeDiseases1

Materials. Acrylamide, bisacrylamide, SDS, Tris, ammonium persulfate, sucrose, and Coomassie Brillant Blue R-250 were obtained from Bio-Rad Laboratories, Richmond, Calif. EDTA and 2-mercaptoethanol were obtained from Sigma Chemical Co., St. Louis, Mo. Hanks' balanced salt solution was obtained from the Grand Island Biological Co., Grand Island, N. Y. Ficoll was obtained from Pharmacia Fine Chemicals, Piscataway, N. J., and Hypaque (sodium 50%) was from Winthrop Laborato ries, New York, N. Y. Patients. Only patients with the leukemic form of their dis ease were studied; i.e. , those who had WBC greater than 10,000 cu mm and/or greater than 50% abnormal cells in the differential counts. The malignant cells were studied from 8 patients with HCL, 8 patients with CLL, 7 patients with ALL, 4 patients with the leukemic phase of PDL, and 1 patient with each of the following: T-ML, ML-LB, and AMoL. The exact diagnosis was confirmed by cytochemistry and lymph node or bone core biopsy. Peripheral blood was collected in a plastic syringe and transferred to a sterile test tube containing EDTA as an anti coagulant. Mononuclear cells were then separated by the Ficoll-Hypaque density gradient method (2). The cells were washed 3 times in Hanks' balanced salt solution and counted. The cell suspension was adjusted to 10 x 106 cells/mI with the Hanks' solution. Immunological characterization of surface immunoglobulins and sheep erythrocyte rosettes was deter mined using methods in our laboratory as described previously (6). Aliquots containing 50 x 106 cells were subsequently pelleted and frozen at —80°. For 1 HCL patient, frozen malignant cells from 2 different days were compared to each other and to freshly prepared cells from one of the same days as to the pattern of protein banding in both SDS-polyacrylamide tube gels and in SDS polyacrylamide gradient slab gels.