Determination of human peripheral blood lymphocyte subpopulations was performed by the use of mixed rosette assay, by which every lymphocyte is able to be classified into a sheep erythrocyte(SRBC)rosette-forming cell or T cell population, an immunobead rosette-forming cell or B cell population, a both cell population and a null cell population. SRBC rosette-negative lymphocytes have been demonstrated to be divided into two populations by direct immunofluorescence; those with the presence of membrane-intrinsic or membrane-stable immuno-globulin(Ig)molecules, and those with membrane-labile IgG molecules which are removed by the incubation at 37℃ before staining. Although immunobeads could react with membrane-stable Ig molecules, but not either with membrane-labile IgG molecules or with Fc receptors, the preincubation of lymphocyte preparations at 37℃ was necessary for the precise determination of T cells and null cells. That is, immune complex or any Ig that might be passively absorbed to the Fc receptors of some of the T cells could inhibit the rosette-formation with SRBC. So far, our study with this mixed rosette assay method disclosed that the mean values and their standard errors of T cells, B cells, both cells, and null cells in peripheral blood lymphocytes obtained from 20 normal subjects were 71.2±2.4%, 12.3±1.1%, 2.3±0.4%, and 14.2±2.1%, respectively. Further study was done in order to well define the characteristics of null cell population. This populaiton lacked the readily detectable C3 receptors which are present of B cells, and every null cell had dense receptors for the Fc portion of IgG. Thus, it seems that null cells determined in this study are equivalent to L lymphocytes.
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