Relationship between mouse lymphocyte receptors for peanut agglutinin (PNA) and Helix pomatia agglutinin (HPA)

Abstract

The relationship between mouse lymphocyte receptors for peanut agglutinin (PNA) and Helix pomatia agglutinin (HPA) has been investigated by immunofluorescence (cocapping) and radiolabeling. In neuraminidase-treated and untreated thymocytes there are two groups of glycoproteins which bind roughly equivalent amounts of PNA. One group also carries all the detectable receptors for HPA, the other binds only PNA. Binding inhibition experiments suggest that PNA and HPA receptors are in close proximity on the shared glycoproteins. The same two groups of receptors are present on 35-40% of neuraminidase-treated spleen lymphoid cells, mainly immunoglobulin (Ig)-negative lymphocytes. Almost all B cells have only PNA-specific receptors. Five-12% of the untreated spleen cells appreciably bind PNA and only a few bind HPA. Solubilized glycoproteins specific for PNA or HPA were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. The major PNA-specific radioiodinated glycoproteins of neuraminidase-treated thymocytes, as isolated by affinity chromatography, consist of the 185-kDa and 195-kDa components of the T200 antigen and of two (diffuse) components of about 140 and 120-125 kDa. All these molecules also bind to HPA-Sepharose, with the exception of the 185 kDa component, which is probably the main constituent of the "pure" PNA receptors on the intact thymocytes. In gels directly labeled with radioactive lectins, the only band strongly labeled by PNA and HPA is the diffuse 140-kDa band. The band at 120 kDa is well labeled by PNA, but all the other components are weakly labeled. The mobility of the 140- and 120-kDa bands depends strongly on neuraminidase-treatment. These bands cannot be detected in gels of untreated thymocytes, but a major HPA-and PNA-specific band of lower molecular weight can be labeled after treating the gels with neuraminidase. The factors determining the differences in labeling pattern obtained by different methods as well as the nature of PNA and HPA binding sites are discussed. The same major PNA- and HPA-binding glycoproteins (apart from minor differences) are present on neuraminidase-treated Ig-negative spleen lymphocytes. The major PNA-binding protein of B lymphocytes appears to correspond to the 225-kDa ("B220") antigen specific for these cells.

Spontaneous redistribution of cell-surface glycoproteins in lymphoid cells during cytokinesis.

“…The ferritin molecules were mainly dispersed as groups of one or two molecules on the nuclear poles ( Figure 3a) and were concentrated in clusters in the cleavage furrow, with larger patches in central region (Figure 3b,c). This distribution is similar to that observed on the uropod of spontaneously capping cells (de Petris and Baumgartner, 1982). No special structures could be identified in this area except for a 60-70 nm cortical microfilament layer similar to that present in the uropods, and, more internally, arrays of microtubules (not shown), some of which apparently terminate at the furrow (Figure 3b).…”

“…One of its visible manifestations is probably the membrane shrinkage and the accumulation of membraneattached particles into the furrow, well documented in classical studies of cytokinesis (see, for example, reviews in Dan, 1948;Wolpert, 1960). As shown by the distribution of HPA-ferritin [which on glutaraldehyde-fixed cells probably mostly labels the glycophorin-like glycoprotein (de Petris and Takacs, 1983;de Petris, 1984)], the HPA receptors remaining on the nuclear poles are mainly dispersed, whereas those transported to the cleavage furrow are aggregated in clusters. This is in agreement with the previous observation that these molecules have a tendency to aggregate spontaneously, and suggests that the aggregates, rather than the single molecules, are preferentially redistributed.…”

“…Particular surface glycoproteins can redistribute and cap 'spontaneously' (i.e., in the absence of external ligands) on the uropod of morphologically polarized leucocytes. Examples of such behaviour are membrane immunoglobulins (m-Ig) on B lymphocytes (Schreiner et al, 1976) and common receptors for the lectins Helix pomatia agglutinin (HPA) and peanut agglutinin (PNA) on mouse and rat thymocytes, T lymphocytes and granulocytes (de Petris and Baumgartner, 1982). During the course of the investigation on the spontaneous capping of HPA and PNA receptors, I observed that in dividing thymocytes the receptors concentrate spontaneously over the cleavage furrow.…”

The glycosylation status of murin postnatal thymus: a study by histochemistry and lectin blotting

“…It has been shown that CD43, CD45 (Favero et al 1984;De Maio et al 1986a, b;De Petris and Takacs 1983;Amado et al 2004) and CD8 (Wu et al 1997) are predominantly PNA binding glycoproteins expressed on immature thymocytes. The CD45 molecule (leukocyte common antigen or T200) is a high molecular weight glycoprotein expressed on cells of hematopoietic origin (Thomas 1989;Charbonneau et al 1988;Holmes 2005) and possesses multiple isoforms ranging between 180 and 230 kDa (Clark and Ledbetter 1989).…”

Isolation and characterization of the potential receptor for wheat germ agglutinin from human neutrophils

“…By selective agglutination with peanut agglutinin (Arachis hypogaea), it is possible to purify cortical immature thymocytes [3]; T and B splenocytes have been fractionated with the lectin from Glycine max [4]. Helix pomatia can be employed for the identification and isolation of T cells [5], and Vicia villosa agglutinin recognizes specifically lymphocytes bearing the CD8+ (cytotoxic) phenotype [6]. Moreover, some of these lectins are currently used to evaluate the immune status of patients [1].…”