A C-type lectin from Bothrops jararacussu venom can adhere to extracellular matrix proteins and induce the rolling of leukocytes

Abstract

Purification of a lectin from Bothrops jararacussu venom (BjcuL) was carried out using agarose-D-galactose affinity gel. MALDI-TOF gave a major signal at m/z 32028, suggesting the presence of a dimmer composed of two identical subunits. Divalent cations were required for the lectin activity, as complete absence of such ions reduced hemagglutination. BjcuL was more effective at neutral pH and showed total loss of activity at pH values below 4.0 and above 9.0. Its agglutinating activity remained stable at 25°C until 60min, but increased when at 35°C for at least 15min. Adhesion assays to extracellular matrix (ECM) glycoproteins showed that the biotinylated lectin (0.039-5.0µg/100µl) was capable of binding to fibronectin and vitronectin in a dose-dependent manner. The binding was partially inhibited in the presence of D-galactose. BjcuL (1.25-10µg/30µl) potential was investigated for leukocyte rolling and adhesion to endothelial cells in living microvessels using intravital microscopy, which showed that it induced a dose-dependent increase in rolling and adherence of leukocytes, acting directly on endothelial cells of postcapillary venules. The specific association between lectins and their ligands, either on the cell surface or on the ECM, is related to a variety of biological processes. The complementary characterization of BjcuL, shown here, is useful to further understand the venom effects and as a background for future investigation for therapeutic strategies

Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom

“…In murine models it is capable of producing edema, increasing vascular permeability [10] and directly inducing cellular infiltration in endothelial cells of post capillary venules, creating an adhesive surface for leukocyte rolling [11]. All of these reports indicate that BJcuL may be involved in the inflammatory process and the activation of immune responses; however, these immunological properties and biological activities are still unclear.…”

BJcuL, a lectin purified from Bothrops jararacussu venom, induces apoptosis in human gastric carcinoma cells accompanied by inhibition of cell adhesion and actin cytoskeleton disassembly

“…When biologically assayed, BJcuL has showed to promote edema and increased vascular permeability in the hind paws of mice (Panunto et al, 2006), which may induce a direct cellular infiltration by creating an adhesive surface for leukocyte rolling (Elifio-Esposito et al, 2007). Recently, we have shown that BJcuL could recognize glycoligands on the neutrophil surface and induce potent human neutrophils activation observed by polarization, migration and adhesion to extracellular matrix (ECM).…”

“…In the present work BJcuL was purified from the crude venom by affinity chromatography on an agarose-D-galactose column as previously described (Elifio-Esposito et al, 2007) and biotinylated according to the manufacturer's recommendations. Using biotinyl-BJcuL, we observed that the lectin was evenly bound on the human gastric carcinoma MKN45 and AGS cells surface (Fig.…”

Lectin isolated from Bothrops jararacussu venom induces IL-10 release by TCD4+ cells and TNF-α release by monocytes and natural killer cells

“…and type I collagen. 6 BjcuL is involved in a wide variety of biological activities including triggering of proinflammatory responses characterized by cellular infiltration into postcapillary venules, 10 increased oedema and vascular permeability in murine models, 11 induced in vitro neutrophil activation, 12 and modulated macrophage functional activation toward an M1 state. 13 Moreover, BjcuL disrupts microbial biofilm structures 14 and induces apoptosis.…”