epiglucan and Gram-Positive-Bacterial-Infections

Pattern recognition receptors (PRRs) are biosensor proteins that bind to non-self pathogen associated molecular patterns (PAMPs). β-1,3-glucan recognition proteins (βGRPs) play an essential role in immune recognition and signaling pathway of insect innate immunity. Here, we report the cloning and characterization of cDNA of OfβGRP3 from Ostrinia furnacalis larvae. The OfβGRP3 contains 1455 bp open reading frame, encoding a predicted 484 amino acid residue protein. In hemocytes, the expression levels of OfβGRP3 in Escherichia coli-challenged group were higher than those of Bacillus subtilis-challenged group at 2, 4, 8, 10 and 12 h post injection (HPI). In fat body, OfβGRP3 expression in both B. subtilis and E. coli-challenged group was significantly higher than that in untreated group from 4 to 10 HPI, and then the expression continuously dropped from 12 to 36 HPI. The OfβGRP3 expression in laminarin-injected group was higher than that in lipopolysaccharides (LPS)-injected group in various test tissues from 4 to 24 HPI. The LT

Topics: Animals; Bacillus subtilis; beta-Glucans; Cells, Cultured; Cloning, Molecular; Gram-Positive Bacterial Infections; Guanine Nucleotide Exchange Factors; Immunity, Innate; Insect Proteins; Larva; Lipopolysaccharides; Moths; Pathogen-Associated Molecular Pattern Molecules; Receptors, Pattern Recognition; Signal Transduction

Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 --> 3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p < 0.05) induced cytokine production IL-6 > TNF alpha > IL-1 beta > GM-CSF > IL-10 > IFN gamma. The induction of these cytokines was significantly (p < 0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P < 0.05) LPS and SA induced cytokines. PTx further augmented (p > 0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti-inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent.

Topics: beta-Glucans; Cell Culture Techniques; Cytokines; Gram-Negative Bacterial Infections; Gram-Positive Bacterial Infections; GTP-Binding Proteins; Humans; Inflammation; Lipopolysaccharides; Monocytes; Proteoglycans; Shock, Septic; Signal Transduction; Staphylococcal Infections; Staphylococcus aureus