A new surface marker on equine peripheral blood lymphocytes. II. Characterization and separation of purified blood lymphocytes with receptors for Helix pomatia A hemagglutinin (HP)

Abstract

In a preceding report we have shown that the lectin Helix pomatia A hemagglutinin (HP) binds to two subpopulations of neuraminidase-treated equine peripheral blood lymphocytes (PBL), constituting about 20% and 75% of PBL, respectively. The aim of the present study was to further characterize these HP+ cells in regard to other surface markers such as receptors for guinea pig erythrocytes (GPR+ cells), membrane-bound immunoglobulins (sIg+ cells), receptors for activated complement (C3R+ cells) and receptors for IgG (Fc alpha R+ cells). This was done by double marker analysis and by lymphocyte fractionation either on columns charged with HP coupled to Sepharose beads or by rosetting with guinea pig erythrocytes. The fractions were also analysed for their proliferative response in the mixed lymphocyte tumor cell interaction (MLTC) assay and to the mitogens leucoagglutinin (La) and concanavalin A (Con A). The results revealed that the majority of GPR+ cells also expressed high avidity receptors for HP, as defined by means of direct immunofluorescence. These cells constituted a subpopulation of GPR+/HP+ cells T cells comprising approximately 20% of PBL. In contrast, about 75% of the HP+ cells in indirect immunofluorescence were GPR-. The fractionation experiments showed that HP+ and GPR+ cells were probably not B cells since they were sIg-. The C3R+ and Fc alpha R+ lymphocytes were heterogeneous in regard to HP receptors but the majority of these cells was also found in the fractions depleted of HP+ and GPR+ lymphocytes. The fractions eluted from HP columns gave a strong proliferative response in MLTC, whereas fractions depleted of HP+ cells responded poorly. However, in contrast to the GPR+-depleted fractions, those enriched in GPR+ lymphocytes responded poorly to the T cell mitogens La and Con A. The mitogenic response of the HP-column fractions to La and to Con A was variable. The results are discussed in relation to HP being a surface marker for a heterogeneous population of equine T cells.