Role of different protein tyrosine kinases in fMLP-induced neutrophil transmigration.
Protein tyrosine phosphorylation is among the early signaling events in polymorphonuclear leukocyte (PMN) responses to chemoattractant stimulation. We previously showed that tyrosine phosphorylation might serve as the downstream signaling for the modulation of PMN transmigration by CD47. Here, we further investigated the role of various tyrosine kinases in PMN transmigration and identified the potential tyrosine kinases serving as CD47-mediated signaling downstream. We observed that PMN transmigration was significantly enhanced by Src family kinase inhibitors PP1 and PP2 as well as Syk tyrosine kinase inhibitor piceatannol, suggesting that these kinases have negative regulatory roles in PMN chemotaxis. In contrast, PMN chemotaxis was reduced by LFM-A13, an inhibitor of the Tec family tyrosine kinase Btk (Bruton's tyrosine kinase). LFM-A13 also dose-dependently inhibited N-formyl-Met-Leu-Phe (fMLP)-induced PMN intracellular [Ca2+] increase. Since LFM-A13 significantly enhanced PMN chemokinesis while other inhibitors had no effect, the inhibition of PMN chemotaxis by LFM-A13 might be due to the promotion of random cell migration. Among the other inhibitors we tested, AG126 significantly inhibited PMN transmigration while the MAP kinase inhibitors SB20358 and PD98059 showed an enhancing effect. No effect of herbimycin A, erbstatin analog, lavendustin A or AG490 on PMN transmigration was observed. Treatment with PP1, PP2 or piceatannol all partially reversed the delay of PMN transmigration caused by inhibitory anti-CD47 antibody. In summary, our results demonstrate distinct roles of different tyrosine kinases in regulating PMN chemotaxis and suggest Src and/or Syk kinases are likely involved in CD47-mediated downstream signaling.
Zen
Ke
K
Cellular and Molecular Biology and Physiology, Department of Biology, Georgia State University, Atlanta, GA 30303, USA. biokxz@langate.gsu.edu
Liu
Yuan
Y
D000577:Amides / Q000494:pharmacology; D051928:CD47 Antigen / Q000378:metabolism*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects / Q000469:parasitology* / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction
D000577:Amides; D051928:CD47 Antigen; C118451:LFM A13; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011505:Protein-Tyrosine Kinases