| Abstract: | Expression of the common acute lymphoblastic leukemia antigen (CALLA) on the surface of individual cells of the human lymphoblastoid lines CW678, Namalwa, and Nalm-6, and the distribution of the antigen epitopes within the cell populations have been determined quantitatively with the murine monoclonal anti-CALLA antibody J5. The distribution of CALLA epitopes in the cell populations was analyzed by indirect immunofluorescence measured by using flow cytometry. The average number of CALLA epitopes per cell were measured by two assays: in a direct assay by binding 125I-labeled antibody J5 to cells, and indirectly by binding 125I-labeled protein A from Staphylococcus aureus to J5-coated cells. On average, CW678, Namalwa, and Nalm-6 cells bore about 1 X 10(4), 6 X 10(4), and 8 X 10(4) CALLA epitopes per cell respectively. Histograms of the absolute number of CALLA epitopes expressed by individual cells in the populations of CW678, Namalwa, and Nalm-6 cultures were generated by a combined analysis of all the binding data. This is the first example of histograms showing quantitative distribution of antigen epitopes. Previously, the expression of antigens by individual cells as obtained by flow cytometry was only presented in terms of relative fluorescence intensity of individual cells in cell populations. |
