Differential Induction of Macrophage GSIB4-Binding Activity

Abstract

Activated macrophages display a terminal galactopyranosyl group on their membrane surface that binds the lectin Griffonia simplicifolia-IB4 (GSIB4). Using FITC-conjugated GSIB4, we examined the induction and subsequent expression of this corresponding marker on peritoneal macrophages from normal (NMO) and LPS-treated (LPS MO) mice. Although the percentage of fresh LPS MO explants that bound GSIB4 was always higher when compared to the NMO counterparts, marker expression on the latter was readily enhanced by culturing the cells in vitro either alone or with stimuli. Moreover, we found that an increase in this activity was promoted by either nonspecific phagocytosis of latex beads, or gamma-interferon (gamma-IFN) treatment. Further investigation showed that a prerequisite sequence of signal delivery to the macrophages was associated with maximal expression of the GSIB4 binding. When gamma-IFN treatment preceded latex bead ingestion, maximum GSIB4 binding occurred. Data obtained from using short-term (1 hr) and long-term (24 hr) exposure to latex beads showed that metabolic processing of induction signals was required to enhance the response over time. This yielded better GSIB4-binding activity when responses to these pulses were analyzed in freshly explanted macrophages. The overall results of this study demonstrated that macrophage binding of GSIB4 was differentially associated with stimuli induction. Moreover, select signals in the form of soluble mediators, or the mechanical events characteristic of internalization were capable of eliciting an increase in the percentage of macrophages that were positive for binding GSIB4. Thus, the enhanced affinity for binding this lectin may serve as a useful marker to determine the magnitude of macrophage responsiveness when these cells are examined following their exposure to different stimuli.

Catecholamines augment collateral vessel growth and angiogenesis in hindlimb ischemia

“…Other studies have used this lectin to determine capillary density in angiogenesis (37,54,86). Although GSL-1-B 4 can stain activated tissue macrophages (88), we would expect their number to have declined by 21 days to a density much lower than muscle capillary density, because there was no evidence of resting ischemia at this time. Consistent with this interpretation, capillary densities before and 21 days after ligation (Fig.…”

Innate Immune Responses to Lung-Stage Helminth Infection Induce Alternatively Activated Alveolar Macrophages

“…An immunohistocytochemical approach was utilized to determine the dynamics of AAMs in the lungs of WT and SCID mice. Alveolar macrophages were identified in histological sections by their ability to bind GSL I, a lectin that binds selectively to ␣-galactose residues on the surfaces of alveolar macrophages (61,66) (Fig. 5A).…”

The Concurrent Expression of Griffonia simplicifolia-IB4 Binding and Tumor Necrosis Factor- Differs between Alveolar and Peritoneal Macrophages

“…GSIB 4-positive cells were recovered by dismantling the columns and immersing the resin in 50 roM a-D-galactose (Sigma), a competitive inhibitory sugar, for 30 min with vigorous agitation (250 rpm). The GSIB 4-positive or -negative status of both populations was verified by immunofluorescent microscopy as performed previously by Tabor et al (15,22). Statistical Analysis.…”

“…This procedure was performed as described previously by 352 DIFFERENCES IN MACROPHAGE MARKERS Tabor et al (15,22). Briefly, macrophages were fixed on coverslips for 30 min with buffered paraformaldehyde and washed twice with warm PBS reaction buffer.…”

“…The first was the ability of these cells to bind the lectin Griffonia simplicifolia-iis; (GSIB 4 ) . This moiety is specific for interacting with membrane (\'-0galactose (14) and is associated with activated, rather than resident, macrophages (15). The second marker was the expression of tumor necrosis factor (TNF)-(\', which is elevated in activated macrophages.…”