Enumeration of human lymphocyte subpopulations by immunofluorescence: a comparative study using automated flow microfluorometry and fluorescence microscopy

These studies reveal that the enumeration of peripheral blood mononuclear cells by fluorescence microscopy and automated flow microfluorometry show a high degree of correlation whether the cells came from normals, individuals with common variable immunodeficiency, chronic lymphocytic leukemia of B cell origin or chronic lymphocytic leukemia of T cell origin. There was excellent agreement between these two methods when counting positive cells stained by the pan-T monoclonal antibodies OKT3 and Leu-1, the helper T reagents OKT4 and Leu-3a, and the suppressor T antibodies OKT8 and Leu-2a. The values obtained for B cells using a pan-B (HB-2) cell antibody analyzed by fluorescence microscopy and automated flow microfluorometry gave a correlation coefficient of 0.86. The percentage of cells identified 3y antibodies with reactivities toward peripheral blood monocytes (MMA or Leu-M1), the HLA-DR determinant, and HNK-1 (Leu-7) positive cells gave correlation coefficients of 0.90, 0.90, and 0.80 respectively when compared by the 2 methods mentioned above. These data suggest that comparable values for lymphocyte subpopulations in human blood samples can be obtained using the most convenient and available technology.