%O Department of Microbiology, College of Veterinary Medicine, King Faisal University, Al-Ahsa, Saudi Arabia; Department of Biological Sciences, College of Science, King Faisal University, Al-Ahsa31982, Saudi Arabia
%R 10.54203/SCIL.2023.WVJ23
%L eprints806
%T NETosis and Calcium influx in Dromedary Camel Neutrophils after In Vitro Toll-like Receptor Stimulation
%J World's Veterinary Journal
%X Neutrophilic granulocytes are vital immune cells of the early response to pathogens. They contribute to the antimicrobial response through phagocytosis, production of reactive oxygen species, cytokine production, degranulation, and NET-formation. Neutrophil extracellular traps (NETs), also known as NETosis, are a critical antibacterial effector mechanism of cells of myeloid effector cells, including neutrophils and macrophages. Toll-like receptors (TLRs) are pattern recognition receptors (PRRs) that mediate pathogen sensing through the recognition of microbial structures known as pathogen-associated molecular patterns (PAMPs). The present study aimed to investigate the potential of several TLR ligands that mimic the sensing of bacterial and viral pathogens to stimulate NET-formation or Ca2+ influx in camel neutrophils. Neutrophils were purified from blood and were stimulated in vitro with ligands to TLR4, TLR2/1, TLR7/8, or TLR3. Net-formation was analyzed using the DNA-sensitive dye SYTOX� Green and staining with antibodies to the neutrophil's granular enzyme myeloperoxidase. Real-time stimulation-induced Ca2+ influx was measured using the Ca2+-sensitive dye Flou-4 and flow cytometry. Only the TLR4-ligand lipopolysaccharide (LPS) could induce NET-formation in camel neutrophils, while none of the investigated TLR agonists showed a Ca2+ influx-inducing effect in camel neutrophils. The current study represents the first report on the impact of direct activation of TLR on NET-formation and Ca2+ influx in camel neutrophils with a selective effect of LPS on NET-formation induction. Future studies may investigate the molecular mechanisms behind the different responsiveness of bovine and camel neutrophils to TLR stimulation. © 2023, World''s Veterinary Journal. All Rights Reserved.
%K lipopolysaccharide; myeloperoxidase; toll like receptor; toll like receptor 1; toll like receptor 2; toll like receptor 3; toll like receptor 4; toll like receptor 7; toll like receptor 8, animal cell; animal experiment; animal model; Article; blood sampling; bovine; calcium transport; camel; cell culture; controlled study; flow cytometry; hemolysis; in vitro study; leukocyte; male; neutrophil; neutrophil extracellular trap; nonhuman; peripheral blood mononuclear cell
%A K. Albahrani
%A J. Alessa
%A B. Falemban
%A M.A. Alkuwayti
%A J. Hussen
%P 214-221
%V 13
%I Scienceline Publication, Ltd
%D 2023
%N 1