IgE receptors on human lymphocytes. III. Expression of IgE receptors on mitogen-stimulated human mononuclear cells.

Human peripheral blood mononuclear cells (PBMC) were tested for the expression of Fc epsilon-receptor (Fc epsilon R) after stimulation with various mitogens in the absence of IgE. Fc epsilon R were found on virtually all the cells from 19 Epstein-Barr virus-transformed B cell lines including those derived from cord blood, from one agamma-globulinemic patient and VDS-0 pre-B cells. Hence, the data clearly indicate that Fc epsilon R may be expressed on very immature B cells. PBMC cultures stimulated with either pokeweed mitogen (PWM), phytohemagglutinin (PHA) or concanavalin A displayed an early increase of their content in Fc epsilon R-bearing cells followed by a decrease to levels below those of control cultures. After fractionation of the PWM-stimulated cultures into T and B cell-enriched preparations, most of the Fc epsilon R+ cells were in the in the B cell fractions and the same low levels of Fc epsilon R+ cells were found in the T cell fractions isolated from the PWM-stimulated and from the control cultures. Double-labeling experiments, employing biotinylated F(ab')2 monoclonal antibody to FcR and either fluorescein isothiocyanate-conjugated B1 or Mo2 monoclonal antibodies, indicated that PWM mainly exerted its effect on B cells and on monocytes. This effect was T cell dependent and it was mediated by soluble factors of T cell origin. At the peak of the PHA or concanavalin A response, most of the Fc epsilon R-bearing cells were found in the B cell fraction but the T cells isolated from mitogen-stimulated cultures contained significant more Fc epsilon R+ cells than those from the control cultures, suggesting that T cell mitogens had increased the expression of Fc epsilon R on some T cells. This view was supported by the finding of a higher proportion of Fc epsilon R+ cells in PHA-stimulated than in control cultures of highly purified T cells with a maximum response at the end of the culture period. Double-labeling experiments at the peak (day 2) of the peripheral blood mononuclear cell response indicated that the expression of Fc epsilon R was increased on B cells (B1+) and on monocytes (Mo2+). By using the same approach at the peak of the T cell response (day 7), it was found that T cells isolated from PHA-stimulated cultures expressed more Fc epsilon R than those isolated from control cultures.(ABSTRACT TRUNCATED AT 400 WORDS)

IgE receptors on human lymphocytes. II. Detection of cells bearing IgE receptors in unstimulated mononuclear cells by means of a monoclonal antibody.

A monoclonal antibody (mAb) specific for lymphocyte IgE receptors (ER) was employed in a rosette assay for the detection of cells bearing IgE receptors (Fc epsilon R). The specificity of the assay was documented by inhibition studies with soluble immunoglobulins (Ig) and anti-Ig antibodies. Moreover, similar results were obtained by employing the F(ab')2 fragment of mAbER instead of intact molecule. Circulating mononuclear cells isolated from normal or allergic adults and from umbilical cord blood contained approximately 8% of Fc epsilon R-bearing cells with values ranging from 0.3 to 17%. Tonsillar lymphocytes contained about 30% of Fc epsilon R+ cells. After the removal of adherent cells, there was a small but significant reduction of the proportion of Fc epsilon R+ cells. When mononuclear cells were separated into T and B cell fractions by two-cycle rosetting with 2-aminoethylisothiouronium bromide hydrobromide-treated sheep red blood cells, most of the Fc epsilon R+ cells were in the B cell fraction; however, a small proportion of Fc epsilon R+ was also found in the enriched T cells and double-labeling experiments confirmed that these cells were indeed T lymphocytes. Fc epsilon R+ cells were purified by rosetting with mAbER-coated erythrocytes and their phenotype was compared to that of Fc epsilon R- cells; Fc epsilon R+ cells contained about 90% of B cells (B1+) together with a small proportion of OKT3+, Leu 7+ and Mo2+ cells. The bulk of T cells, macrophages and natural killer (NK) cells was found in the Fc epsilon R- cells which contained fewer B cells than the fraction of Fc epsilon R+ cells. These data thus indicated that the great majority of Fc epsilon R-bearing cells are B cells but that a small proportion of NK cells, macrophages and T lymphocytes also express Fc epsilon R. Upon incubation at 37 degrees C, B cells lost their Fc epsilon R and this phenomenon was selectively inhibited by IgE; however, purified T cells seemed to express more Fc epsilon R after overnight incubation at 37 degrees C and this was not influenced by IgE. It is finally shown that the expression of Fc epsilon R is cyclic and that Fc epsilon R-bearing B cells do not represent a functionally distinct subpopulation of B lymphocytes.