Abstract
Considering the significance of mast cells (MCs) in the course of various physiological and pathological processes, and the pivotal role of endogenous molecules, i.e., cathelicidins and defensins as multifunctional modulators, the study examines the constitutive and cathelicidin LL-37/defensin hBD-2-induced expression of certain NLRs and RLRs, i.e., NOD1, NOD2, and RIG-I, in fully-mature tissue MCs, and the impact of LL-37 and hBD-2 on MC pro-inflammatory activity. All experiments were carried out in vitro on freshly-isolated peritoneal (P)MCs. qRT-PCR, western blotting, flow cytometry, and confocal microscopy were used to evaluate both constitutive and LL-37/hBD-2-induced expression of NOD1, NOD2, and RIG-I receptors. ROS was determined using H2DCFDA, and Boyden microchamber assay was used to define the migratory response. Standard techniques assessed histamine, cysLT, and chemokine generation. PMCs express NOD1, NOD2, and RIG-I constitutively. LL-37 and hBD-2 enhance the expression and induce translocation of the studied receptors and directly activate the pro-inflammatory and migratory responses of PMCs. Observations demonstrate that LL-37 and hBD-2 might augment MC capability and sensitivity to NLR and RLR ligands and strengthen the role of MCs in inflammation.
Highlights
Host defense peptides (HDPs) are evolutionarily-conserved biologically-active molecules synthesized as the first line of defense by a variety of organisms
The qRT-PCR and western blotting were used to determine the expression of NOD1, NOD2, and retinoic acid-inducible gene I (RIG-I) mRNAs and proteins by native PMCs, and those stimulated with LL-37 or hBD-2
We investigated whether PMCs express NOD1, NOD2, and RIG-I proteins and whether LL-37 and hBD-2 have an impact on their baseline levels
Summary
Host defense peptides (HDPs) are evolutionarily-conserved biologically-active molecules synthesized as the first line of defense by a variety of organisms. Cathelicidins and defensins have multidimensional properties allowing them to act directly against bacterial[3], viral[4] and fungal[5] invasion but new research has cast light on alternative functionalities, including immunomodulatory activities They can serve as chemokines, and stimulate the production and release of various immunoregulatory mediators by inflammatory cells; they are recognized as potent agents in inflammatory processes. MCs constitute the chief sentinels of the immune system, with a multiplicity of functions in the maintenance of a range of physiological features They are considered crucial for the regulation of body homeostasis by acting on wound healing, angiogenesis, and vascular permeability, as well as taking part in the homeostasis of tissues and organs undergoing continuous growth and remodeling. It discusses the effects of LL-37 and hBD-2 on PMC pro-inflammatory activity
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