Production of TNF-α, IL-1β, IL-12 and IFN-γ in murine peritoneal macrophages on treatment with wheat germ agglutinin in vitro: involvement of tyrosine kinase pathways

Abstract

Treatment of macrophages with various doses of wheat germ agglutinin (WGA) for different time intervals resulted in enhanced expression of TNF-a, IL-1b, IL-12 and IFN-g. The maximum expressions were observed at 24 h with 100 ng/ml of WGA. Enhanced transcription of cytokines TNF-a, IL-1b, IL-12, and IFN-g was observed at 16 h of WGA treatment by RT-PCR. Pharmacological inhibitor of tyrosine kinase, PI3 kinase, protein kinase C, p42/44, p38, JNK and intracellular calcium immobilizing agent down regulated the WGA induced expression of cytokines TNF-a, IL-1b, IL-12 and IFN-g. Maximum protein tyrosine kinase activity in macrophages was seen at 5 min of WGA treatment. Maximum cytosolic Ca ++ was observed at 10 min of WGA treatment. WGA treated macrophages showed maximum activation of protein kinase C (PKC) and PI3 kinase at 10 min, p42/44, p38 at 15 min and JNK at 30 min. Transcription factor ELK1 was activated at 60 min and IêB, c-Fos and c-Jun at 30 min of WGA treatment. The pharmacological inhibitors were also used to check the cascade of activation of tyrosine kinase, PKC, PI3 kinase, p42/44, p38, JNK and release of calcium from intracellular storage to sort out the signal pathways involved in the release of TNF-a, IL-1b, IL-12, and IFN-g by macrophages on treatment with WGA in vitro.

Recognition of TLR2 N-Glycans: Critical Role in ArtinM Immunomodulatory Activity

“…Stimulation of murine macrophages with ArtinM also enhanced TLR2 expression on the cell surface, a result that is compatible with the finding that Concanavalin A (Con A) induces higher TLR2–9 expression in murine peritoneal macrophages [43], [44]. TLR2 expression is differentially regulated by inflammatory mediators [45]–[48], and some infections have been associated with increased TLR2 expression [49]–[54].…”

“…It was previously reported that peritoneal macrophages stimulated with wheat germ agglutinin (WGA) produce proinflammatory cytokines such as TNF-α, IL-1β, IL-12, and IFN-γ in a manner that is dose- and time-dependent, as well as sensitive to different pharmacological inhibitors (p38 MAPK, PI3K, JNK, and ERK1/2). In addition, IL-1β production by macrophages stimulated with ConA occurs through the PI3K, PKC, ERK 1/2, p38 MAPK, and JAK/STAT pathways [43]. These data suggest that lectins with different specificities may induce macrophage activation through several signaling pathways by targeting different glycosylated receptors.…”

The western diet and lifestyle and diseases of civilization

“…26,100,154 Moreover, lectins from lentils, kidney beans, peas, and wheat potently increase the production of inflammatory cytokines (interleukin [IL]-12, IL-2, and interferon γ) in cell cultures, 253 and WGA also stimulates production of tumor necrosis factor (TNF)-α and IL-1β in vitro. 254 Furthermore, WGA and PHA induce the production of metalloproteinases (MMPs) in leukocytes, 255,256 and WGA directly causes the activation of platelets and potently increases their aggregation. 257 This may be relevant because rupture of the fibrous cap and formation of the blood clot, which is mediated by MMPs and platelets, is a crucial mechanism involved in thrombus production.…”

The Dietary Intake of Wheat and other Cereal Grains and Their Role in Inflammation

“…IL-12, in turn, activated the secretion of IFN-γ by T or natural killer cells [52]. In murine peritoneal macrophages WGA induced the production of the pro-inflammatory cytokines TNF-α, IL-1β, IL-12 and IFN-γ [53]. Similar results have been observed in isolated human PBMC, given that nanomolar concentrations of WGA stimulated the release of several pro-inflammatory cytokines.…”