The role of cyclic nucleotides in neutrophil migration (original) (raw)
Related papers
Inflammation Research, 1997
Objective: To study the effect of cyclic nucleotides and PDE-resistant cyclic nucleotide analogues on neutrophil migration. Methods: Migration of electropermeabilized neutrophils in Boyden chambers in vitro. Results: Addition of cyclic AMP inhibited migration of electropermeabilized neutrophils in the presence of cGMP, relative to the level of migration in the presence of cGMP alone. However, when cGMP was replaced with 8-pCPT-cGMP or Sp-cGMPS, analogues of cGMP which are not degraded by phosphodiesterases, cAMP synergistically enhanced migration. In contrast, migration in the presence of the phosphodiesterase-resistant cAMP analogue, Sp-cAMPS, was not enhanced by addition of cGMP. Conclusions: Taking into account reports in the literature which show that cGMP-hydrolysing activity can be enhanced by the catalytic subunit of cAMP-dependent protein kinase, it is hypothesized that breakdown of cGMP in neutrophils may be modulated via cAMP.
The effect of cyclic GMP and cyclic AMP on migration by electroporated human neutrophils
European Journal of Pharmacology: …, 1993
Human neutrophils retain the ability to migrate when they are electroporated; this enables the study of potential mediators by direct application to the cell interior. Cyclic GMP strongly enhances random migration by electroporated human neutrophils. The effect is maximal at a concentration of 10 tzM. The potentiating effect of cGMP is chemokinetic. Chemotaxis of electroporated neutrophils activated by formylmethionylleucylphenylalanine (fMLP) is stimulated by cGMP at concentrations up to 10 tzM; higher concentrations inhibit chemotaxis. Cyclic AMP resembles cGMP in that both activation and inhibition may occur. However, activation occurs over a very small concentration range, and inhibition is a predominant feature. Cyclic nucleotide-activated migration is dependent on Ca 2÷, in contrast with activation of migration by fMLP.
British Journal of Pharmacology, 1997
The aim of this study was to establish the role of nitric oxide (NO) and cyclic GMP in chemotaxis and superoxide anion generation (SAG) by human neutrophils, by use of selective inhibitors of NO and cyclic GMP pathways. In addition, inhibition of neutrophil chemotaxis by NO releasing compounds and increases in neutrophil nitrate/nitrite and cyclic GMP levels were examined. The ultimate aim of this work was to resolve the paradox that NO both activates and inhibits human neutrophils. 2 A role for NO as a mediator of N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced chemotaxis was supported by the ®nding that the NO synthase (NOS) inhibitor L-NMMA (500 mM) inhibited chemotaxis; EC 50 for fMLP 28.76+5.62 and 41.13+4.77 pmol/10 6 cells with and without L-NMMA, respectively. Similarly the NO scavenger carboxy-PTIO (100 mM) inhibited chemotaxis; EC 50 for fMLP 19.71+4.23 and 31.68+8.50 pmol/10 6 cells with and without carboxy-PTIO, respectively. 3 A role for cyclic GMP as a mediator of chemotaxis was supported by the ®nding that the guanylyl cyclase inhibitor LY 83583 (100 mM) completely inhibited chemotaxis and suppressed the maximal response; EC 50 for fMLP 32.53+11.18 and 85.21+15.14 pmol/10 6 cells with and without LY 83583, respectively. The same pattern of inhibition was observed with the G-kinase inhibitor KT 5823 (10 mM); EC 50 for fMLP 32.16+11.35 and 4135 pmol/10 6 cells with and without KT 5823, respectively. 4 The phosphatase inhibitor, 2,3-diphosphoglyceric acid (DPG) (100 mM) which inhibits phospholipase D, attenuated fMLP-induced chemotaxis; EC 50 for fMLP 19.15+4.36 and 61.52+16.2 pmol/10 6 cells with and without DPG, respectively. 5 Although the NOS inhibitors L-NMMA and L-canavanine (500 mM) failed to inhibit fMLP-induced SAG, carboxy-PTIO caused signi®cant inhibition (EC 50 for fMLP 36.15+7.43 and 86.31+14.06 nM and reduced the maximal response from 22.14+1.5 to 9.8+1.6 nmol O 2 7 /10 6 cells/10 min with and without carboxy-PTIO, respectively). This suggests NO is a mediator of fMLP-induced SAG. 6 A role for cyclic GMP as a mediator of SAG was supported by the eects of G-kinase inhibitors KT 5823 (10 mM) and Rp-8-pCPT-cGMPS (100 mM) which inhibited SAG giving EC 50 for fMLP of 36.26+8.77 and 200.01+43.26 nM with and without KT 5823, and 28.35+10.8 and 49.25+16.79 nM with and without Rp-8-pCTP-cGMPS. 7 The phosphatase inhibitor DPG (500 mM) inhibited SAG; EC 50 for fMLP 33.93+4.23 and 61.12+14.43 nM with and without DPG, respectively. 8 The NO releasing compounds inhibited fMLP-induced chemotaxis with a rank order of potency of GEA 3162 (IC 50 =14.72+1.6 mM)4GEA 5024 (IC 50 =18.44+0.43 mM)4SIN-1 (IC 50 41000 mM). This order of potency correlated with their ability to increase cyclic GMP levels rather than the release of NO, where SIN-1 was most eective (SIN-1 (EC 50 =37.62+0.9 mM)4GEA 3162 (EC 50 =39.7+0.53 mM)4 GEA 5024 (EC 50 =89.86+1.62 mM)). 9 In conclusion, chemotaxis and SAG induced by fMLP can be attenuated by inhibitors of phospholipase D, NO and cyclic GMP, suggesting a role for these agents in neutrophil activation. However, the increases in cyclic GMP and NO induced by fMLP, which are associated with neutrophil activation, are very small. In contrast much larger increases in NO and cyclic GMP, as observed with NO releasing compounds, inhibit chemotaxis.
Inflammation, 1997
Preincubation of human neutrophils with aurothiomalate had little effect on random migration or chemotactic migration towards the chemotactic peptide fMLP. However, a strong enhancement of migration was observed when aurothiomalate was present in a concentration gradient; the effect of the drug was chemotactic rather than chemokinetic. Thiomalate also caused a chemotactic enhancement of migration but here a tenfold higher concentration of the drug was required as compared with aurothiomalate. Aurothiomalate caused an increase of cellular cGMP level, and inhibitors of guanylate cyclase inhibited the activating effect of aurothiomalate. Three specific antagonists of cGMP-dependent kinase (G-kinase) strongly inhibited aurothiomalate-induced migration of electroporated neutrophils. The results suggest that stimulation of migration by aurothiomalate is mediated by cGMP and a G-kinase. Monoclonal anti-IL-8 inhibited aurothiomalate-induced stimulation of migration. Though no increased release of IL-8 could be established upon exposure of neutrophils to aurothiomalate, it seems conceivable that the stimulating effect of aurothiomalate is mediated by IL-8.
British Journal of Pharmacology, 1997
Angiotensin II had a bimodal effect on human neutrophil migration. Low concentrations of angiotensin II stimulated random migration. At a concentration of 10−10M it caused a maximal increase of migration; migration increased from 47.2±2.1 μm in the absence of angiotensin II, to 73.1±2.2 μm with 10−10M angiotensin II present in the lower compartment of the Boyden chamber (n=5, P<0.001). Stimulation of migration by angiotensin II was partly chemotactic and partly chemokinetic. Angiotensin II concentrations of 10−8M and higher inhibited chemotactic peptide-stimulated chemotaxis.The stimulant effect of angiotensin II on migration was completely dependent on extracellular Ca2+. In the presence of 1 mM Ca2+, angiotensin II stimulated migration to 76.1±1.7 μm, while migration in the absence of Ca2+ was 42.2±1.9 μm (n=4, P<0.001). Different types of calcium channel blockers either moderately or strongly inhibited angiotensin II-activated migration. Stimulation of migration by angiotensin II in intact cells required higher concentrations of Ca2+ than in electroporated cells. This supports the view that there is an influx of Ca2+ through the plasma membrane, and a requirement of calcium for an intracellular target.Angiotensin II-stimulated migration was inhibited by pertussis toxin; from 71.6±2.0 μm in the absence, to 43.6±1.5 μm in the presence of pertussis toxin (n=4, P<0.001). Migration of electroporated neutrophils stimulated by angiotensin II was synergistically enhanced by GTPγS. This suggests that one or more G-proteins are involved in the activating effect of angiotensin II.Inhibitors of soluble guanylate cyclase and antagonists of cyclic GMP-dependent kinase strongly inhibited the activating effect of angiotensin II. The results suggest that the activating effect of angiotensin II is mediated by cyclic GMP and by cyclic GMP-dependent kinase.Angiotensin II had a bimodal effect on human neutrophil migration. Low concentrations of angiotensin II stimulated random migration. At a concentration of 10−10M it caused a maximal increase of migration; migration increased from 47.2±2.1 μm in the absence of angiotensin II, to 73.1±2.2 μm with 10−10M angiotensin II present in the lower compartment of the Boyden chamber (n=5, P<0.001). Stimulation of migration by angiotensin II was partly chemotactic and partly chemokinetic. Angiotensin II concentrations of 10−8M and higher inhibited chemotactic peptide-stimulated chemotaxis.The stimulant effect of angiotensin II on migration was completely dependent on extracellular Ca2+. In the presence of 1 mM Ca2+, angiotensin II stimulated migration to 76.1±1.7 μm, while migration in the absence of Ca2+ was 42.2±1.9 μm (n=4, P<0.001). Different types of calcium channel blockers either moderately or strongly inhibited angiotensin II-activated migration. Stimulation of migration by angiotensin II in intact cells required higher concentrations of Ca2+ than in electroporated cells. This supports the view that there is an influx of Ca2+ through the plasma membrane, and a requirement of calcium for an intracellular target.Angiotensin II-stimulated migration was inhibited by pertussis toxin; from 71.6±2.0 μm in the absence, to 43.6±1.5 μm in the presence of pertussis toxin (n=4, P<0.001). Migration of electroporated neutrophils stimulated by angiotensin II was synergistically enhanced by GTPγS. This suggests that one or more G-proteins are involved in the activating effect of angiotensin II.Inhibitors of soluble guanylate cyclase and antagonists of cyclic GMP-dependent kinase strongly inhibited the activating effect of angiotensin II. The results suggest that the activating effect of angiotensin II is mediated by cyclic GMP and by cyclic GMP-dependent kinase.British Journal of Pharmacology (1997) 121, 643–648; doi:10.1038/sj.bjp.0701167
The effect of pentoxifylline on human neutrophil migration: A possible role for cyclic nucleotides
Biochemical Pharmacology, 1997
Relatively low concentrations of pentoxihlline caused a stimulation of random migration, while high concentrations inhibited chemotactic migration activated by formyl-methionyl-leucyl-phenylalanine (fMLP). The stimulating effect of pentoxyfylline was partly chemokinetic and partly chemotactic, and was dependent on extracellular calcium. Activation of migration by pentoxifylline was not dependent on the pore size of the micropore filter, indicating that the effect was not mediated by the ability of the drug to induce membrane deformability. Inhibitors of guanylate cyclase and antagonists of cGMP-dependent protein kinase (G-kinase) inhibited stimulation of migration by pentoxifylline. Pentoxyfylline caused a transient increase in cGMP level, while only high concentrations of pentoxifylline caused an increase in cyclic adenosine monophosphate (CAMP) level. It is suggested that the increase of migration is caused by cGMP and is mediated by a G-kinase, while the inhibition of migration at high concentrations of pentoxifylline is mediated by CAMP.
N-acetylcysteine causes a transient stimulation of neutrophil migration
Immunopharmacology, 1998
Ž. Random migration of rabbit peritoneal neutrophils was enhanced in a chemokinetic way by N-acetylcysteine NAC in a Ž. small concentration range 10-400 mM. The enhancement was due to the cysteine moiety in the molecule, because cysteine equally caused a stimulation of random migration. The stimulating effect of NAC or cysteine largely disappeared when cells were preincubated with NAC or cysteine for 30 min before submission to chemotaxis, indicating that desensitization occurs. The stimulating effect of NAC was dependent on extracellular calcium. Because the Ca 2q-dependence of migration by electroporated cells differed from that of intact cells, and because calcium channel blockers inhibited the effect of NAC, the calcium-dependent target is probably located inside the cell rather than on the cell surface. In contrast with fMLP, NAC did not cause an upregulation of CD11b expression of cells in suspension. Inhibitors of guanylate cyclase and of cGMP-depen-Ž. dent protein kinase G-kinase inhibited stimulation of migration by NAC, suggesting that cGMP played a decisive role in the stimulatory effect of NAC. q 1998 Elsevier Science B.V.
Immunopharmacology, 1979
The possible involvement in vitro of 3', t'-cyclic GMP (cGMP) in the mechanism of action of the lymphokine, leukocyte migration inhibitory factor (LIF), was investigated. Partially purified LIF-rich supernatants, but not their control counterparts, induced a 2-fold increase in the cGMP levels of purified human polymorphonuclear (NMN) leukocytes. The effect was no influenced by heat-inactivated horse serum; it was manifested within 3 min of exposure to LIF and it subsided within 180 min. LIF and the supernatant factor responsible for the cGMP-generating effect were both rendered inactive by treatment with the serine esterase and protease inhibitor, phenylmethylsulfonyl fluoride, indicating that these factors are closely related, if not identical. A potent phosphodiesterase inhibitor, dipyridamole (2 x 10(-4) M), induced a 3- to 5-fold increase in PMN leukocyte cGMP levels, but combined treatment with purified LIF and dipyridamole did not add to this effect. This suggests that...
Atrial natriuretic factor stimulates migration by human neutrophils
European Journal of Pharmacology: Molecular Pharmacology, 1995
Atrial natriuretic factor causes a strong stimulation of human neutrophil migration in the concentration range of 4 x 10 9 and 10 -7 M. The effect, which depends on the presence of extracellular Mg 2+ but not on extracellular Ca 2+, is composed of a chemokinetic and a chemotactic component. Cyclic GMP level of neutrophils is enhanced by atrial natriuretic factor. Two inhibitors of soluble guanylate cyclase, 6-anilino-5,8-quinolinedione (LY 83583) and methylene blue, have no effect on stimulation of migration by atrial natriuretic factor. Atrial natriuretic factor-activated migration is inhibited by pertussis toxin. Migration by electroporated neutrophils is synergistically enhanced by guanosine-5'-[3,thio]triphosphate (GTP3,[S]) and atrial natriuretic factor or by GTP3,[S] and chemotactic peptide, while GTP3,[S] and dioctanoyl glycerol give an additive effect. The results suggest that besides a modulation via cGMP a part of the effect of atrial natriuretic factor on migration is regulated via the ANF receptor-subtype that does not activate guanylate cyclase.
The effect of endothelin-2 (ET2) on migration and changes in cytosolic free calcium of neutrophils
Naunyn-schmiedebergs Archives of Pharmacology, 1996
The effect of endothelin-2 (ET-2) on neutrophil migration and intracellular calcium was studied. Depending on the concentration, ET-2 enhanced or inhibited neutrophil migration. At low concentrations ET-2 caused a chemotactic stimulation of migration, in contrast with endothelin-1 (ET-1) which caused a chemokinetic stimulation of migration. At higher concentrations ET-2 inhibited formyl-methionylleucyl-phenylalanine(fMLP)-activated migration. Both activation and inhibition by ET-2 were completely dependent on extracellular Ca2+. Unlike ET-1 which caused an increase in cytosolic free Ca2+ at a concentration which stimulated migration, ET-2 caused a measurable increase of cytosolic free Ca2+ at a concentration which did not stimulate migration. This strongly suggests that there is no correlation between maximal stimulation of cytoplasmic free calcium, and maximal stimulation of migration. Influx of extracellular Ca2+ was required for both activation of migration and change in cytosolic free Ca+, because no effect was observed in the absence of extracellular Ca+, and because blockers of Ca2+-influx inhibited ET-2-activated migration. The ETA-receptor antagonist cyclo(-D-Trp-D-Asp-Pro-D-Val-Leu) (BQ123), and the ETB-receptor antagonist [Cys11-Cys15]-endothelin-1(11–21) (IRL1038) antagonized the stimulatory effect of ET-2 on migration, and the inhibitory effect of high concentrations of ET-2 on fMLP-activated chemotaxis. This suggests that both the ETA-receptor and the ETB-receptor are involved in the stimulatory effect of low concentrations of ET-2, and in the inhibitory effect of high concentrations of ET-2.