Microphotometric differentiation of human T and B cells tagged with monospecific immunoadsorbent beads

A comparative and statistical study was done of the classification of peripheral blood lymphocytes (PBLs) as T cells, B cells and monocytes by various immunologic procedures and computerized microphotometric analysis. PBLs from 15 healthy male subjects were examined by immunofluorescence, E rosettes, and immunoadsorbent beads (IAB) for T cells and B cells, by phase contrast microscopy and as fixed slide preparations. Cells tagged with IAB for T cells and B cells were fixed, stained with Papanicolaou stain and analyzed. Evaluation of immunologic data shows 50% to 57% T cells, 9.7% to 24.1% B cells, 13.4% to 16.1% monocytes and about 20% unmarked cells. Analysis of T cells shows significant correlations between E and T cell-IAB rosettes, but neither rosetting procedure reveals a positive correlation with immunofluorescence-labelled T cells. Comparison of B cells shows an insignificant correlation between B-cell IAB and immunofluorescence. Results showed that permanent slides of rosetted cells can be made without alteration in relative numbers of rosetted cells. Assessment of immunologically tagged cell samples by image analysis correctly classified 80% to 90% of cells as T and B cells. Evaluation of homogeneity of the T and B cell populations shows the existence of four subsets of B cells and five subsets of T cells.