LYMPHOCYTE SUBSET DETERMINATION USING A HEMATOLOGY ANALYZER

Anti-CD4 antibody (T4)-coated microspheres were used to label CD4 cell s in whole blood, The mixture was lysed and analyzed by a modified Cou lter VCS hematology analyzer, which differentiated microsphere-labeled cells by a change in Coulter volume, conductance, and light scatter. %CD3(+)/CD4(+) fluorescent values from a profile were compared to %CD4 values using the VCS-microsphere method. CD3 gating was used to exclu de CD4(+) monocytes from the 90LS-FALS lymphocyte gate. The results co rrelated well (R = 0.996). The percentage of CD4(+) lymphocytes from p rofile scatterplots and VCS scatterplots showed a line of regression c lose to the equivalence line (n = 76, slope = 0.96) when CD3 gating wa s used for the profile. These results suggest that CD3 gating, though necessary for SOLS-FALS scatterplots, may not be necessary for volume- conductance-light scatterplots. (C) 1995 Wiley-Liss, Inc.