Activation of kinin B1 receptors induces chemotaxis of human neutrophils

Abstract

Kinins are biologically active peptides that are powerful mediators of cellular inflammation. They mimic the cardinal signs of inflammation by inducing vasodilatation and by increasing vascular permeability and pain. Neutrophils are chemoattracted to sites of inflammation by several stimuli. However, the evidence concerning the chemotactic effect of kinin peptides has been contradictory. We analyzed the chemotactic effect of kinin B(1) receptor agonists on neutrophils isolated from peripheral blood of human healthy subjects. Chemotaxis was performed using the migration under agarose technique. To test the effect of B(1) receptor agonists, each assay was carried out overnight at 37 degrees C in 5% CO(2)-95% air on neutrophils primed with 1 ng/ml interleukin-1beta. Simultaneous experiments were performed using unprimed cells or cells challenged with formyl-Met-Leu-Phe (fMLP). A clear chemotactic activity was observed when primed neutrophils were challenged with Lys-des[Arg(9)]-bradykinin (LDBK) or des[Arg(9)]-bradykinin at 10(-10) M but not when unprimed cells were used. A reduction in the chemotactic response was observed after priming of cells in the presence of 0.5 mM cycloheximide and 10 mug/ml brefeldin A, suggesting that some protein biosynthesis is required. Techniques such as reverse transcriptase-polymerase chain reaction and in situ hybridization confirmed the expression of the B(1) receptor mRNA, and immunocytochemistry and autoradiography demonstrated the expression of the B(1) receptor protein. In contrast to other chemoattractants such as fMLP, cytosolic intracellular calcium did not increase in response to the B(1) receptor agonist LDBK. A generation of kinin B(1) receptor agonists during the early phase of acute inflammation may favor the recruitment of neutrophils to the inflammatory site.

Inhibitory effect of plant phenolics on fMLP-induced intracellular calcium rise and chemiluminescence of human polymorphonuclear leukocytes and their chemotactic activityin vitro

“…Induction of directional movement of PMNs without any stimulatory effect on other cell functions is not a unique property among the chemoattractant substances. For instance, bradykinin and innate defense-regulator peptide (IDR-1) both attracted human PMNs, while the first compound simply had no effect on [Ca 2+ ] i , the second compound had no effect on both [Ca 2+ ] i and superoxide formation (Ehrenfeld et al, 2006;Lee & Bae, 2008). Likewise, some phenolics can selectively switch on the transmission of signals leading to intracellular actin reorganization and induction of PMNs migration; however, elucidation of this activity requires further studies.…”

Signaling Pathways Coupled to Activation of the Kinin B1 Receptor

“…Furthermore, the treatment of the hemagglutinin-tagged human B1R, expressed in HEK293 cells, with Nglycosidase F resulted in the conversion of the receptor from a heterogeneous specie migrating at 35 to 45 kDa to a relatively homogeneous one migrating at 37 kDa. We have observed a similar electrophoretic pattern of the B1R protein in human neutrophils (Ehrenfeld et al, 2006), breast cancer cells (Molina et al, 2009), endothelial cells and fibroblasts (our group, unpublished data). It is postulated that glycosylation probably increases the hydrophilicity of the extracellular portions of B1R affecting or regulating ligand affinity, efficient G protein coupling, maturation (folding, stabilization), intracellular trafficking and receptor oligomerization or receptor degradation (Menke et al, 1994;MacNeil et al, 1995;Leeb-Lundberg et al, 2005).…”

“…In contrast, activation of B1R induces the migration of human PC3 prostate cancer cells via activation of focal adhesion kinase (FAK), an important kinase involved in cytoskeletal reorganization and cell migration (Taub et al, 2003). On the other hand, studies performed in human neutrophils show that B1R stimulates leukocyte chemotaxis, though the pathways implicated in this process are still unknown (Ahluwalia & Perretti, 1996;Paegelow et al, 2002;Ehrenfeld et al, 2006). Moreover, it is known that release of the kinin moiety from kininogens sited on the neutrophil surface by enzymatic action of the classical tissue kallikrein (hK1) results in opening of junctions between the endothelial cells, thereby causing plasma extravasation (Stuardo et al, 2004).…”

“…In vascular smooth muscle cells, B1R stimulation produces a significant dependence on extracellular Ca 2+ and a transient increase in phosphatidilinositol hydrolysis that is more sustained than that generated by the B2R (Tropea et al, 1993). Nevertheless, in the rat renal cortical collecting duct cells, keratinocytes, breast cancer cells and neutrophils B1R does not induce intracellular Ca 2+ mobilization (Ehrenfeld et al, 2006;Matus et al, 2008;Molina et al, 2009) Another characteristic of B1R is its capacity to regulate the cell cycle. In some cases, B1R is proliferative as seen in fibroblasts, vascular smooth muscle cells, estrogen-sensitive breast cancer cells, and this response involves MAPK activation (Marceau and Tremblay, 1986;Christopher et al, 2001;Molina et al, 2009).…”

Fever-like temperature modification differentially affects in vitro signaling of bradykinin B1 and B2 receptors

“…Moreover, unlike B 2 R, B 1 R does not become desensitized and responds in some cell types to activation with an increase in surface receptor number rather than internalization, e.g., in IMR 90 cells (Phagoo et al, 2000;Leeb-Lundberg et al, 2005). As both kinin receptors participate in inflammatory processes by mediating the release of proinflammatory cytokines and recruitment of immune cells (McLean et al, 2000;Leeb-Lundberg et al, 2005;Ehrenfeld et al, 2006), the properties of the B 1 R indicate sustained signaling and thus a role in the prolonged phase of the immune response with amplification of inflammatory processes (Blaukat et al, 1996;Austin et al, 1997;Marceau et al, 1998;Faussner et al, 1999;Phagoo et al, 2000;Marceau et al, 2002;Leeb-Lundberg et al, 2005). Inflammation is characterized by five cardinal symptoms: pain, redness, swelling, loss of function, and heat (Elliott et al, 1960).…”