Flow cytometric analysis of the binding of eleven lectins to human T‐ and B‐cells and to human T‐ and B‐cell lines

Abstract

The relative surface binding of 11 lectins to human peripheral blood T‐ and B‐lymphocytes, to Molt‐4 and JM T‐cell lines, and to 6410 and NC37 B‐cell lines was determined by flow cytometry. The lectins from Lens culinaris (LCA), Ricinus communis (RCA), Arachis hypogaea (PNA), Abrus precatorius (APA), Ulex europaeus (UEA‐F), Sarothamnus scoparius (SAS‐F), Helix pomatia (HPA), Phaseolus coccineus (L‐PHA), Glycine max (SBA), and Triticum vulgare (WGA) were fluoresceinated and incubated with living, formaldehyde‐fixed, or neuraminidase‐treated cells. Except LCA, which preferentially bound to the two B‐cell lines tested in this study, none of the other lectins exhibited selective binding to the undifferentiated cells of the cell lines. The T‐cell lines and, in part, the peripheral blood T‐cells bound less WGA, APA, LCA, and L‐PHA than the B‐cell lines and the peripheral blood B‐cells. Binding of PNA was found only after neuraminidase treatment of the cells; the binding of PNA, HPA, and UEA‐F after neuraminidase treatment was higher for the T‐cells than the B‐cells from peripheral blood. No significant differences were detected between both cell types for RCA, ConA, SBA, and SAS‐F.

Detection of avian macrophages with concanavalin a

“…Flow cytometric analysis of lectin binding has been used to detect cell surface changes, to determine lymphocyte subpopulations under normal and pathological conditions, and to determine the stage of differentiation or development of certain cells of mammalian species (Madewell et al, 1984;Malin-Berdel et al, 1984). The present results demonstrate that these techniques can be applied to the avian system with consistent results.…”

“…These analyses have been performed on both cell suspensions and tissue sections with assays including cell agglutination, histochemical staining, and, more recently, flow cytometric analysis (Madewell et al, 1984;Malin-Berdel et al, 1984. In a previous study (Greenfield et al, 1986), histochemical staining with peroxidase labelled concanavalin A (con A) was used to detect a pathological change in a small population of leukocytes within the bursa of Fabricus. With infectious bursal disease virus (IBDV) infection, cells that appeared to be macrophages were found to rearrange into foci within the bursa.…”

Characterization of lectin isolated from Momordica charantia seed as a B cell activator

“…Additionally, using flow cytometric analysis, Malin-Berdel et al reported on the relative surface binding of 11 lectins (including ConA) to human peripheral blood T and B cells and human T-and B-cell lines. Only lectin from Lens culinaris (LCA), which preferentially bound to the two Bcell lines, exhibited selective binding to the undifferentiated cells [15]. Malin-Berdel et al [15] postulated that the conflicting results possibly reflect the influence of cell volume on the evaluation of cell-bound fluorescence [16] or may be due to the use of much higher lectin concentrations [17] and different reaction conditions that make additional binding sites accessible [18].…”

Lectin Binding Assays for In-Process Monitoring of Sialylation in Protein Production

“…Lectins are carbohydrate-binding proteins which have been extensively used for detection and characterization of glycoproteins. Lectin binding is not only specific to individual sugar structure, but also appears to depend on local steric interactions in the immediate molecular neighborhood of the target carbohydrates [24,33,35]. Lectins have been used to characterize cell surface glycans because of their specificity in terms of branching, linkage and terminal structure of glycans [36,37].…”