Delineation and quantitation of human peripheral blood lymphocyte subpopulations by electrophoretic mobility and role of surface charge in cell to cell interaction

Abstract

A modified procedure is proposed for determination of electrophoretic mobility (EPM) of lymphocytes which enables readings on 200 cells from a sample size of 5 x 10⁶ lymphocytes. In normal human peripheral blood, B lymphocytes (bearing surface immunoglobulin and receptor for activated C₃) were found to carry the lowest charge (0.94 ± 0.05 μ/sec/V/cm). Their number on the EPM basis was 16.9 ± 4.2%. Two subpopulations of T lymphocytes, one carrying high charge (T_H) with a mean EPM of 1.26 ± 0.04 μ /sec/V/cm and the other carrying lower charge (T_L) with a mean EPM of 1.11 ± 0.03 μ /sec/V/cm were discernible. The former subpopulation of T cells formed spontaneous rosettes with SRBC in 10 min and the latter represented the remaining T lymphocytes which formed rosettes with 24 hr of incubation with SRBC. The number of early rosette-forming cells increased from 69 to 89% when T cells were incubated with neuraminidase-treated SRBC. These observations suggest that complementary electrostatic charge on T cells and SRBC are among the forces facilitating formation of rosettes between these cells.