| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
*
Kidney Disease Program and
Department of Biochemistry and Molecular Biology, University of Louisville and Veterans Affairs Medical Center, Louisville, KY 40292; and
Franz Volhard Clinic at the Max Delbrück Center for Molecular Medicine, Virchow-Klinikum-Charité, Humboldt University of Berlin, Berlin, Germany
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We studied the effect of GM-CSF on constitutive PMN apoptosis after in
vitro culture in the presence of 10% FCS. We report that GM-CSF
inhibits PMN spontaneous apoptosis by 
65%. Pharmacologic inhibition
of the ERK pathway and PI 3-kinase significantly attenuated
GM-CSF-induced apoptosis delay. GM-CSF caused rapid phosphorylation of
Akt and BAD proteins. To determine whether the PI 3-kinase and ERK
pathways are used by other agonists, we examined their role in
IL-8-dependent delay of constitutive PMN apoptosis. Pharmacologic
inhibition of the ERK and PI 3-kinase pathways resulted in attenuation
of IL-8-induced apoptosis delay. PI 3-kinase and ERK activation play a
key role in the delay of PMN apoptosis by the inflammatory cytokines
GM-CSF and IL-8.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Endotoxin-free reagents and plastic disposables were used in all experiments. Recombinant human IL-8 and recombinant human GM-CSF were obtained from Genzyme (Cambridge, MA). The ERK inhibitor PD98059 was purchased from Biomol (Plymouth Meeting, PA) and the PI 3-kinase inhibitors wortmannin and LY294002 were obtained from Calbiochem (San Diego, CA). Anti-Akt and anti-phospho-Akt Abs were obtained from New England Biolabs (Beverly, MA). Anti-BAD and anti-phospho-BAD Abs were purchased from Upstate Biotechnology (Lake Placid, NY).
Isolation of PMNs and culture conditions
PMN were isolated from venous blood obtained from healthy
volunteers collected in ACD tubes. Cells were pelleted for 20 min at
175 x g and protein-rich plasma was aspirated.
Protein-rich plasma was further centrifuged for 15 min at 1500 x
g to produce protein-poor plasma (PPP). Erythrocytes were
separated from the cell suspension by dextran sedimentation and the
leukocyte-enriched fraction was centrifuged for 10 min at 175 x
g. Pelleted cells were resuspended in 3 ml PPP and
underlayered with 3 ml each of 42% Percoll in PPP and 50.5% Percoll
in PPP. Following centrifugation for 10 min at 180 x
g, PMNs were aspirated from the 42 to 51% interface and
washed in PPP. Any remaining RBCs were removed by hypotonic lysis and
PMNs were resuspended in cation-free Krebs-Ringer-phosphate solution.
PMN preparations routinely contained >95% PMNs, as determined by
morphology, and were >99% viable. Apoptosis was induced by extended
culture in the presence of medium supplemented with FCS. Briefly, PMNs
were cultured for 18 h in RPMI 1640 supplemented with 10% FCS,
L-glutamine, penicillin, and streptomycin and
placed in a 5% CO2/air incubator. GM-CSF (20
ng/ml) was added to PMN cultures as indicated. In experiments where
kinase inhibitors were used, the inhibitors were added to PMNs for 60
min before the addition of GM-CSF. PD98059, wortmannin, and LY294002
had no effect on cell viability or the rate of constitutive apoptosis
at the indicated concentrations.
Flow cytometric measurement of PMN apoptosis
Flow cytometry was used to measure DNA content at the single-cell level using a modified method of Kettritz et al. (6). The method takes advantage of the fact that activated endonucleases generate low m.w. DNA fragments in apoptotic cells. After membrane permeabilization, these fragments leak out, resulting in decreased DNA content in apoptotic cells, whereas DNA content in nonapoptotic cells remains unchanged. In brief, isolated PMNs were pelleted and in PBS containing 0.5 mM EDTA. Chilled 95% ethanol was added to a final concentration of 70% to permeabilize the cells and the cell mixtures were stored at -20°C for 1–2 days. PMNs were resuspended in a PBS/EDTA/1% BSA solution. DNase-free RNase and propidium iodide were added. Cells were held for 15 min in the dark at room temperature, then incubated for 20 min at 30°C, and then stored at 4°C for 6–8 h in the staining mixture, allowing low molecular DNA fragments to leave permeabilized cells. PMN DNA content was analyzed using a Coulter Epics Profile II flow cytometer (Palo Alto, CA).
Immunoblotting of phosphorylated Akt and BAD
Phosphorylated Akt was identified using a rabbit polyclonal Ab directed against phosphorylated Ser473 (New England Biolabs). Phosphorylated BAD Ab (anti-phospho-Ser136) was obtained from Upstate Biotechnology. Immunoblotting was performed using a method previously described, with modification (19). Briefly, PMNs (2 x 107 cells/ml) were incubated with GM-CSF (20 ng/ml) in the presence and absence of inhibitors for the described time periods. Cells were then pelleted and resuspended in a lysis buffer containing Tris-HCl, NaCl, Triton X-100, Nonidet P-40, EDTA, EGTA, Na2VO4, NaF, aprotinin, leupeptin, and PMSF. The cells were incubated on ice in lysis buffer for 5 min and repelleted, and the supernatants were denatured in Laemmli buffer. Protein samples were then resolved on a 15% SDS-acrylamide gel and transferred to nitrocellulose membranes and washed in a blocking solution containing 5% dried milk in PBS. The nitrocellulose membranes were then incubated overnight in HRP-conjugated primary Ab (anti-phospho-BAD, Ser136, 1:750 dilution, anti-phospho-Akt, 1:150 dilution) in 5% BSA/PBS. The nitrocellulose was then washed and incubated with HRP-conjugated goat anti-rabbit Ab (1:1000 dilution in 5% BSA in PBS). Bands were detected using an enhanced chemiluminescence technique and exposure to photographic film. Autoradiographs were then scanned using a personal densitometer (Molecular Dynamics, Sunnyvale, CA). Equal loading of protein in lanes was confirmed in all experiments by stripping and reprobing the blots for either total BAD or Akt.
Statistical analysis
Statistical analysis was performed using GraphPad Instat software (GraphPad Software, San Diego, CA). ANOVA for repeated measures was performed to detect differences between multiple groups. Differences between groups were determined by use of a Tukey posttest. Data are presented as means ± SEM. Differences were considered to be significant when the p value was < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
PI 3-kinase controls several survival pathways. Therefore, we
examined the role of PI 3-kinase in delay of PMN apoptosis by GM-CSF.
PMNs were pretreated with LY294002, a PI 3-kinase inhibitor, for 1
h before overnight incubation with 20 ng/ml GM-CSF. As shown in Fig. 1
, 10 µM LY294002 partially reversed
the protective effect of GM-CSF. PI 3-kinase inhibition attenuated
GM-CSF-dependent apoptosis delay by 55% (percent PMN apoptosis:
63% untreated vs 41% GM-CSF treated vs 51% GM-CSF + LY294002
apoptosis). Similar inhibition of GM-CSF protection from apoptosis was
observed with 50 µM wortmannin, an alternative inhibitor of PI
3-kinase (data not shown). These data indicate that delay of
spontaneous apoptosis by GM-CSF depends in part on PI 3-kinase
activation.
|
One mechanism used by PI 3-kinase to prevent apoptosis is
activation of the protein kinase B/Akt pathway (3, 20, 21). Fig. 2 is a representative
immunoblot (n = 6) of phosphorylated Akt in
GM-CSF-stimulated PMNs. PMNs were incubated with 20 ng/ml GM-CSF for 1,
2, 4, and 5 min. Minimal phosphorylated Akt was seen at zero time, but
significant Akt phosphorylation was observed after 1 min. PI 3-kinase
or its products can phosphorylate Akt (22). To confirm
that PI 3-kinase controls GM-CSF-dependent Akt phosphorylation in PMNs,
we performed immunoblots of phospho-Akt in the presence of LY294002. As
shown in Fig. 2, PI 3-kinase inhibition by 10 µM LY294002 blocked
GM-CSF-induced Akt phosphorylation. Similar results were obtained with
50 µM wortmannin (data not shown).
|
Phosphorylated Akt can exert an anti-apoptotic effect by, in
turn, phosphorylating the protein BAD. In many cell types, BAD
phosphorylation causes dissociation of BAD from anti-apoptotic
proteins of the Bcl family, resulting in prolonged survival. We
examined whether GM-CSF activation of Akt was associated with
phosphorylation of BAD. Shown in Fig. 3
is a representative immunoblot of phosphorylated BAD (n
= 6). BAD can be phosphorylated at Ser112 and
Ser136 (23, 24). Akt phosphorylates
Ser136 and therefore we performed immunoblots
using Abs against the relevant Ser136
phosphorylation site (23). Immunoblotting of PMNs exposed
to 20 ng/ml GM-CSF for various times was performed. BAD phosphorylation
can occur via PI 3-kinase-dependent and -independent pathways
(24, 25). To determine whether PI 3-kinase activation is
necessary for BAD phosphorylation, we examined the effect of LY294002
on GM-CSF-induced BAD phosphorylation. As shown in Fig. 3A,
10 µM LY294002 significantly attenuated GM-CSF-dependent BAD. Fig. 3B summarizes the pooled data from six separate
experiments.
|
Activation of the ERK pathway also exerts an anti-apoptotic
effect in many cells. Therefore, we used the ERK pathway inhibitor PD
98059 to examine whether ERK played a role in GM-CSF-induced delay of
PMN apoptosis. PMNs were pretreated for 1 h with 50 µM PD98059
and incubated for 18 h with 20 ng/ml GM-CSF. Fifty micromolar
PD98059 alone had no significant effect on the rate of spontaneous
apoptosis. As shown in Fig. 4, 20 ng/ml
GM-CSF decreased the percentage of apoptotic PMNs from 59 to 31%.
PD98059 attenuated the GM-CSF protective effect, resulting in
apoptosis in 48.0 ± 1.2% of the cells, a 61% reversal of
the GM-CSF effect.
|
To determine whether simultaneous inhibition of ERK and PI 3-kinase had an additive effect, we measured apoptosis in GM-CSF-treated PMNs that were pretreated with both PD98059 and LY294002. PMNs were incubated in the presence and absence of GM-CSF (20 ng/ml) with and without the ERK inhibitor PD98059 (50 µM), the PI 3-kinase inhibitor LY294002 (10 µM), or both inhibitors. PMN apoptosis was determined by flow cytometric analysis of DNA content. Combined treatment with PD98059 and LY294002 did not further inhibit GM-CSF apoptosis delay when compared with the effect of each agent (data not shown).
Activation of Akt by IL-8
To determine whether PI 3-kinase and ERK are used by other ligands
that delay neutrophil apoptosis, we examined the role of these pathways
in IL-8-induced apoptosis delay. IL-8 caused rapid phosphorylation of
Akt as shown in Fig. 5, and this was
markedly attenuated by 10 µM LY294002.
|
We examined whether IL-8 activation of Akt was associated with
phosphorylation of BAD. Shown in Fig. 6A is a representative
immunoblot of phosphorylated BAD, and pooled data from six experiments
are shown in B. PMNs were incubated with 100 nM IL-8 for the
times indicated and immunoblots with ant-phospho-BAD Ab were performed.
As shown in Fig. 6, IL-8 rapidly stimulated BAD phosphorylation in
PMNs, and this was markedly attenuated by LY294002.
|
LY294002 also caused reversal of IL-8-dependent apoptosis delay as
demonstrated in Fig. 7A.
Additionally, ERK pathway inhibition by 50 µM PD98059 significantly
attenuated IL-8-induced delay of PMN apoptosis (Fig. 7B).
These data indicate that IL-8 and GM-CSF use similar signaling
mechanisms to inhibit constitutive PMN apoptosis.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Reports in the literature indicate that two inhibitors of tyrosine kinases, herbimycin A and genistein, can attenuate the effect of GM-CSF on PMN apoptosis (22). An additional inhibitor of tyrosine phosphatase, phenylarsine oxide, promotes PMN survival (31). Wei et al. (8) used antisense oligonucleotides to deplete PMNs of specific tyrosine kinases associated with the GM-CSF receptor. Antisense depletion of Lyn kinase, but not Hck kinase, resulted in reversal of GM-CSF-induced apoptosis delay. Nonreceptor-associated tyrosine kinases such as Lyn activate a number of signal transduction pathways that can inhibit apoptosis. First, tyrosine kinase activation can lead to activation of ERK that in turn has been shown in many cells to exert an anti-apoptotic effect (32). Second, tyrosine kinases can activate PI 3-kinase that then can activate PKB/Akt, a potent anti-apoptotic signaling pathway. We observed that inhibition of PI 3-kinase partially reversed the GM-CSF-dependent delay of PMN apoptosis. GM-CSF also activated the PI 3-kinase target Akt that has been shown to inhibit apoptosis in a number of reports (25, 33), and GM-CSF-dependent Akt phosphorylation was blocked by the PI 3-kinase inhibitors LY294002 and wortmannin. Our data indicate that PI 3-kinase activation clearly results in phosphorylation of Akt and phosphorylated Akt may inhibit apoptosis by several actions. First, phosphorylated Akt functions as a serine-threonine kinase and can phosphorylate BAD, a member of the Bcl-2 family. BAD, when not phosphorylated, promotes apoptosis by forming an inhibiting heterodimer with anti-apoptotic members of the Bcl-2 family (34). Phosphorylated BAD dissociates from Bcl-2, thereby increasing the anti-apoptotic effects of the Bcl-2 family. Although the anti-apoptotic Bcl-2-like proteins in PMNs that form heterodimers with BAD are not clearly delineated, our data demonstrate that GM-CSF stimulates BAD phosphorylation. Akt can potentially inhibit apoptosis by two other mechanisms. Phosphorylated Akt causes activation of ERK and Akt has been shown to phosphorylate and inactivate an important member of the cell death pathway, procaspase 9 (35). Therefore, GM-CSF activation of Akt in PMNs could inhibit apoptosis by inactivating caspases, stimulating ERK activity, or disinhibiting anti-apoptotic proteins in the Bcl-2 family.
Several studies have reported that MAPKs play a key role in regulating apoptosis. In general, p38 MAPK and ERK appear to promote and inhibit programmed cell death, respectively (12, 13, 14, 15, 16). Exceptions to this observation have been documented. For example, Frasch et al. (15) observed that neutrophils undergo apoptosis by p38 MAPK-dependent and -independent mechanisms. As noted above, protein tyrosine kinase-dependent pathways can activate ERK, but additional activation pathways have been observed. Several studies have reported that PI 3-kinase is an element in the signal transduction pathway leading to ERK activation (36, 37, 38). We have reported previously that GM-CSF-dependent ERK activation in PMNs is attenuated by wortmannin, a PI 3-kinase inhibitor (39). In the present study, neither ERK nor PI 3-kinase inhibition could fully reverse GM-CSF-dependent apoptosis delay, suggesting an additional mechanism is at work. Additionally, combined PI 3-kinase and ERK inhibition did not completely reverse the effect of GM-CSF on apoptosis. This indicates that these pathways are not activated in parallel or working independently in delaying apoptosis. We have previously reported that GM-CSF activates PMN p38 MAPK and others have observed that p38 MAPK plays a role in constitutive PMN apoptosis (39). Therefore, we examined the effect of SB203580 on GM-CSF-induced apoptosis delay and found no effect (data not shown)
The role of PI 3-kinase and ERK in IL-8-induced apoptosis delay was examined to determine whether the PI 3-kinase and ERK pathways are shared by other agonists that delay PMN apoptosis. The activation of these pathways plays a functional role in some effects of IL-8 on PMNs (38). As observed with GM-CSF, IL-8 stimulated Akt phosphorylation that was blocked by PI 3-kinase inhibitors. PI 3-kinase and ERK pathway inhibitors significantly attenuated IL-8-induced delay of PMN apoptosis. IL-8 also stimulated phosphorylation of BAD at the Ser136 site and this was inhibited by LY294002. Collectively, our data indicate that GM-CSF and IL-8 delay PMN apoptosis, in part, by activating ERK and PI 3-kinase. GM-CSF and IL-8 activation of PMN are associated with stimulation of the PI 3-kinase/Akt/BAD anti-apoptotic pathway. Pharmacologic blockade of the ERK and PI 3-kinase/Akt pathway may potentially exert an anti-inflammatory effect by promoting PMN apoptosis.
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Jon B. Klein, Kidney Disease Program, University of Louisville, 615 South Preston Street, Louisville, KY 40292.
3 Abbreviations used in this paper: PMN, polymorphonuclear neutrophil; MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinase; PI 3-kinase, phosphoinositide 3-kinase; PPP, protein-poor plasma.
Received for publication April 20, 1999. Accepted for publication February 3, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-mediated apoptosis. J. Am. Soc. Nephrol. 8:1091.[Abstract]
-mediated apoptosis of human neutrophils. Kidney Int. 53:84.[Medline]
and GM-CSF. J. Leukocyte Biol. 64:537.[Abstract]
-receptors mediate activation of Akt/protein kinase B in human phagocytes. J. Biol. Chem. 272:28096. stimulates increased plasma membrane guanine
nucleotide binding protein activity in polymorphonuclear leukocytes
[published erratum appears in 1995 J. Leukocyte Biol.
57:803.]. J. Leukocyte Biol. 57:500.
to the protein kinases PKB and MAPK. Science 282:293.RIII) expression. J. Immunol. 153:1254.[Abstract]
-mediated mitogen-activated protein kinase signaling pathway. J. Biol. Chem. 271:12133. and GM-CSF. J. Leukocyte Biol. 64:537.
This article has been cited by other articles:
|
|
 |
|
  A. Paunel-Gorgulu, M. Zornig, T. Logters, J. Altrichter, U. Rabenhorst, J. Cinatl, J. Windolf, and M. Scholz Mcl-1-Mediated Impairment of the Intrinsic Apoptosis Pathway in Circulating Neutrophils from Critically Ill Patients Can Be Overcome by Fas Stimulation J. Immunol., November 15, 2009; 183(10): 6198 - 6206. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-N. Li, P. G. Barlow, J. Bylund, A. Mackellar, A. Bjorstad, J. Conlon, P. S. Hiemstra, C. Haslett, M. Gray, A. J. Simpson, et al. Secondary necrosis of apoptotic neutrophils induced by the human cathelicidin LL-37 is not proinflammatory to phagocytosing macrophages J. Leukoc. Biol., October 1, 2009; 86(4): 891 - 902. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Binet and D. Girard Novel human neutrophil agonistic properties of arsenic trioxide: involvement of p38 mitogen-activated protein kinase and/or c-jun NH2-terminal MAPK but not extracellular signal-regulated kinases-1/2 J. Leukoc. Biol., December 1, 2008; 84(6): 1613 - 1622. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Didichenko, N. Spiegl, T. Brunner, and C. A. Dahinden IL-3 induces a Pim1-dependent antiapoptotic pathway in primary human basophils Blood, November 15, 2008; 112(10): 3949 - 3958. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Himpe, C. Degaillier, A. Coppens, and R. Kooijman Insulin-like growth factor-1 delays Fas-mediated apoptosis in human neutrophils through the phosphatidylinositol-3 kinase pathway J. Endocrinol., October 1, 2008; 199(1): 69 - 80. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. El Kebir, L. Jozsef, and J. G. Filep Opposing regulation of neutrophil apoptosis through the formyl peptide receptor-like 1/lipoxin A4 receptor: implications for resolution of inflammation J. Leukoc. Biol., September 1, 2008; 84(3): 600 - 606. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Pluskota, D. A. Soloviev, D. Szpak, C. Weber, and E. F. Plow Neutrophil Apoptosis: Selective Regulation by Different Ligands of Integrin {alpha}M{beta}2 J. Immunol., September 1, 2008; 181(5): 3609 - 3619. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. El Kebir, L. Jozsef, W. Pan, and J. G. Filep Myeloperoxidase Delays Neutrophil Apoptosis Through CD11b/CD18 Integrins and Prolongs Inflammation Circ. Res., August 15, 2008; 103(4): 352 - 359. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. I. Kasahara, M. E. Poynter, Z. Othman, D. Hemenway, and A. van der Vliet Acrolein Inhalation Suppresses Lipopolysaccharide-Induced Inflammatory Cytokine Production but Does Not Affect Acute Airways Neutrophilia J. Immunol., July 1, 2008; 181(1): 736 - 745. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Uddin, G. Seumois, L. C. Lau, P. Rytila, D. E. Davies, and R. Djukanovic Enhancement of neutrophil function by the bronchial epithelium stimulated by epidermal growth factor Eur. Respir. J., April 1, 2008; 31(4): 714 - 724. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Wu, H. Kausar, P. Johnson, D. E. Montoya-Durango, M. Merchant, and M. J. Rane Hsp27 Regulates Akt Activation and Polymorphonuclear Leukocyte Apoptosis by Scaffolding MK2 to Akt Signal Complex J. Biol. Chem., July 27, 2007; 282(30): 21598 - 21608. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. El Kebir, L. Jozsef, T. Khreiss, W. Pan, N. A. Petasis, C. N. Serhan, and J. G. Filep Aspirin-Triggered Lipoxins Override the Apoptosis-Delaying Action of Serum Amyloid A in Human Neutrophils: A Novel Mechanism for Resolution of Inflammation J. Immunol., July 1, 2007; 179(1): 616 - 622. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Watanabe, W. Matsuyama, Y. Shirahama, H. Mitsuyama, K.-i. Oonakahara, S. Noma, I. Higashimoto, M. Osame, and K. Arimura Dual Effect of AMD3100, a CXCR4 Antagonist, on Bleomycin-Induced Lung Inflammation J. Immunol., May 1, 2007; 178(9): 5888 - 5898. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. D. Wrann, N. A. Tabriz, T. Barkhausen, A. Klos, M. van Griensven, H. C. Pape, D. O. Kendoff, R. Guo, P. A. Ward, C. Krettek, et al. The Phosphatidylinositol 3-Kinase Signaling Pathway Exerts Protective Effects during Sepsis by Controlling C5a-Mediated Activation of Innate Immune Functions J. Immunol., May 1, 2007; 178(9): 5940 - 5948. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Brkovic, M. Pelletier, D. Girard, and M. G. Sirois Angiopoietin chemotactic activities on neutrophils are regulated by PI-3K activation J. Leukoc. Biol., April 1, 2007; 81(4): 1093 - 1101. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. E. Kilpatrick, S. Sun, D. Mackie, F. Baik, H. Li, and H. M. Korchak Regulation of TNF mediated antiapoptotic signaling in human neutrophils: role of {delta}-PKC and ERK1/2 J. Leukoc. Biol., December 1, 2006; 80(6): 1512 - 1521. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R.-F. Guo, L. Sun, H. Gao, K. X. Shi, D. Rittirsch, V. J. Sarma, F. S. Zetoune, and P. A. Ward In vivo regulation of neutrophil apoptosis by C5a during sepsis J. Leukoc. Biol., December 1, 2006; 80(6): 1575 - 1583. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Tortorella, O. Simone, G. Piazzolla, I. Stella, V. Cappiello, and S. Antonaci Role of Phosphoinositide 3-Kinase and Extracellular Signal-Regulated Kinase Pathways in Granulocyte Macrophage-Colony-Stimulating Factor Failure to Delay Fas-Induced Neutrophil Apoptosis in Elderly Humans J. Gerontol. A Biol. Sci. Med. Sci., November 1, 2006; 61(11): 1111 - 1118. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. G. Barlow, Y. Li, T. S. Wilkinson, D. M. E. Bowdish, Y. E. Lau, C. Cosseau, C. Haslett, A. J. Simpson, R. E. W. Hancock, and D. J. Davidson The human cationic host defense peptide LL-37 mediates contrasting effects on apoptotic pathways in different primary cells of the innate immune system J. Leukoc. Biol., September 1, 2006; 80(3): 509 - 520. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Li, D. J. Allendorf, R. Hansen, J. Marroquin, C. Ding, D. E. Cramer, and J. Yan Yeast beta-Glucan Amplifies Phagocyte Killing of iC3b-Opsonized Tumor Cells via Complement Receptor 3-Syk-Phosphatidylinositol 3-Kinase Pathway J. Immunol., August 1, 2006; 177(3): 1661 - 1669. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Rosas, P. F. Dijkers, C. L. Lindemans, J.-W. J. Lammers, L. Koenderman, and P. J. Coffer IL-5-mediated eosinophil survival requires inhibition of GSK-3 and correlates with {beta}-catenin relocalization J. Leukoc. Biol., July 1, 2006; 80(1): 186 - 195. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. A. Lindemans, P. J. Coffer, I. M. M. Schellens, P. M. A. de Graaff, J. L. L. Kimpen, and L. Koenderman Respiratory Syncytial Virus Inhibits Granulocyte Apoptosis through a Phosphatidylinositol 3-Kinase and NF-{kappa}B-Dependent Mechanism J. Immunol., May 1, 2006; 176(9): 5529 - 5537. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Kettritz, M. Choi, B. Salanova, M. Wellner, S. Rolle, and F. C. Luft Fever-Like Temperatures Affect Neutrophil NF-{kappa}B Signaling, Apoptosis, and ANCA-Antigen Expression J. Am. Soc. Nephrol., May 1, 2006; 17(5): 1345 - 1353. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. M. McGettrick, J. M. Lord, K.-Q. Wang, G. E. Rainger, C. D. Buckley, and G. B. Nash Chemokine- and adhesion-dependent survival of neutrophils after transmigration through cytokine-stimulated endothelium J. Leukoc. Biol., April 1, 2006; 79(4): 779 - 788. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Tabary, H. Corvol, E. Boncoeur, K. Chadelat, C. Fitting, J. M. Cavaillon, A. Clement, and J. Jacquot Adherence of airway neutrophils and inflammatory response are increased in CF airway epithelial cell-neutrophil interactions Am J Physiol Lung Cell Mol Physiol, March 1, 2006; 290(3): L588 - L596. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. A. Sawatzky, D. A. Willoughby, P. R. Colville-Nash, and A. G. Rossi The Involvement of the Apoptosis-Modulating Proteins ERK 1/2, Bcl-xL and Bax in the Resolution of Acute Inflammation in Vivo Am. J. Pathol., January 1, 2006; 168(1): 33 - 41. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K.-S. Choi, J. T. Park, and J. S. Dumler Anaplasma phagocytophilum Delay of Neutrophil Apoptosis through the p38 Mitogen-Activated Protein Kinase Signal Pathway Infect. Immun., December 1, 2005; 73(12): 8209 - 8218. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. D. Kobayashi, J. M. Voyich, A. R. Whitney, and F. R. DeLeo Spontaneous neutrophil apoptosis and regulation of cell survival by granulocyte macrophage-colony stimulating factor J. Leukoc. Biol., December 1, 2005; 78(6): 1408 - 1418. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. von Gunten, S. Yousefi, M. Seitz, S. M. Jakob, T. Schaffner, R. Seger, J. Takala, P. M. Villiger, and H.-U. Simon Siglec-9 transduces apoptotic and nonapoptotic death signals into neutrophils depending on the proinflammatory cytokine environment Blood, August 15, 2005; 106(4): 1423 - 1431. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Sakamoto, F. Hato, T. Kato, C. Sakamoto, M. Akahori, M. Hino, and S. Kitagawa Type I and type II interferons delay human neutrophil apoptosis via activation of STAT3 and up-regulation of cellular inhibitor of apoptosis 2 J. Leukoc. Biol., July 1, 2005; 78(1): 301 - 309. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Bruno, S. Conus, I. Schmid, and H.-U. Simon Apoptotic Pathways Are Inhibited by Leptin Receptor Activation in Neutrophils J. Immunol., June 15, 2005; 174(12): 8090 - 8096. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. C. Parker, M. K. B. Whyte, S. K. Dower, and I. Sabroe The expression and roles of Toll-like receptors in the biology of the human neutrophil J. Leukoc. Biol., June 1, 2005; 77(6): 886 - 892. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Rane, D. Gozal, W. Butt, E. Gozal, W. M. Pierce Jr, S. Z. Guo, R. Wu, A. D. Goldbart, V. Thongboonkerd, K. R. McLeish, et al. {gamma}-Amino Butyric Acid Type B Receptors Stimulate Neutrophil Chemotaxis during Ischemia-Reperfusion J. Immunol., June 1, 2005; 174(11): 7242 - 7249. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Pinho, D. G. Souza, M. M. Barsante, F. P. Hamer, M. S. De Freitas, A. G. Rossi, and M. M. Teixeira Phosphoinositide-3 kinases critically regulate the recruitment and survival of eosinophils in vivo: importance for the resolution of allergic inflammation J. Leukoc. Biol., May 1, 2005; 77(5): 800 - 810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Machida, H. Inoue, K. Matsumoto, M. Tsuda, S. Fukuyama, H. Koto, H. Aizawa, Y. Kureishi, N. Hara, and Y. Nakanishi Activation of PI3K-Akt pathway mediates antiapoptotic effects of {beta}-adrenergic agonist in airway eosinophils Am J Physiol Lung Cell Mol Physiol, May 1, 2005; 288(5): L860 - L867. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Hu and M. M. Sayeed Activation of PI3-kinase/PKB contributes to delay in neutrophil apoptosis after thermal injury Am J Physiol Cell Physiol, May 1, 2005; 288(5): C1171 - C1178. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Bennouna and E. Y. Denkers Microbial Antigen Triggers Rapid Mobilization of TNF-{alpha} to the Surface of Mouse Neutrophils Transforming Them into Inducers of High-Level Dendritic Cell TNF-{alpha} Production J. Immunol., April 15, 2005; 174(8): 4845 - 4851. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. R. Katsumoto, J. Duda, A. Kim, Z. Wardak, G. Dranoff, D. W. Clapp, and K. Shannon Granulocyte/macrophage colony-stimulating factor and accessory cells modulate radioprotection by purified hematopoietic cells J. Exp. Med., March 21, 2005; 201(6): 853 - 858. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Francois, J. El Benna, P. M. C. Dang, E. Pedruzzi, M.-A. Gougerot-Pocidalo, and C. Elbim Inhibition of Neutrophil Apoptosis by TLR Agonists in Whole Blood: Involvement of the Phosphoinositide 3-Kinase/Akt and NF-{kappa}B Signaling Pathways, Leading to Increased Levels of Mcl-1, A1, and Phosphorylated Bad J. Immunol., March 15, 2005; 174(6): 3633 - 3642. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Walmsley, C. Print, N. Farahi, C. Peyssonnaux, R. S. Johnson, T. Cramer, A. Sobolewski, A. M. Condliffe, A. S. Cowburn, N. Johnson, et al. Hypoxia-induced neutrophil survival is mediated by HIF-1{alpha}-dependent NF-{kappa}B activity J. Exp. Med., January 3, 2005; 201(1): 105 - 115. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kotone-Miyahara, K. Yamashita, K.-K. Lee, S. Yonehara, T. Uchiyama, M. Sasada, and A. Takahashi Short-term delay of Fas-stimulated apoptosis by GM-CSF as a result of temporary suppression of FADD recruitment in neutrophils: evidence implicating phosphatidylinositol 3-kinase and MEK1-ERK1/2 pathways downstream of classical protein kinase C J. Leukoc. Biol., November 1, 2004; 76(5): 1047 - 1056. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Li, R. Shu, G. Filippatos, and B. D. Uhal Apoptosis in lung injury and remodeling J Appl Physiol, October 1, 2004; 97(4): 1535 - 1542. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. E. Kilpatrick, S. Sun, and H. M. Korchak Selective regulation by {delta}-PKC and PI 3-kinase in the assembly of the antiapoptotic TNFR-1 signaling complex in neutrophils Am J Physiol Cell Physiol, September 1, 2004; 287(3): C633 - C642. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Derouet, L. Thomas, A. Cross, R. J. Moots, and S. W. Edwards Granulocyte Macrophage Colony-stimulating Factor Signaling and Proteasome Inhibition Delay Neutrophil Apoptosis by Increasing the Stability of Mcl-1 J. Biol. Chem., June 25, 2004; 279(26): 26915 - 26921. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Bozinovski, J. Jones, S.-J. Beavitt, A. D. Cook, J. A. Hamilton, and G. P. Anderson Innate immune responses to LPS in mouse lung are suppressed and reversed by neutralization of GM-CSF via repression of TLR-4 Am J Physiol Lung Cell Mol Physiol, April 1, 2004; 286(4): L877 - L885. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Kettritz, M. Choi, S. Rolle, M. Wellner, and F. C. Luft Integrins and Cytokines Activate Nuclear Transcription Factor-{kappa}B in Human Neutrophils J. Biol. Chem., January 23, 2004; 279(4): 2657 - 2665. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Paine III, S. E. Wilcoxen, S. B. Morris, C. Sartori, C. E. O. Baleeiro, M. A. Matthay, and P. J. Christensen Transgenic Overexpression of Granulocyte Macrophage-Colony Stimulating Factor in the Lung Prevents Hyperoxic Lung Injury Am. J. Pathol., December 1, 2003; 163(6): 2397 - 2406. [Abstract] [Full Text]
|
|
|
|
|
|
M. Choi, S. Rolle, M. Wellner, M. C. Cardoso, C. Scheidereit, F. C. Luft, and R. Kettritz Inhibition of NF-{kappa}B by a TAT-NEMO-binding domain peptide accelerates constitutive apoptosis and abrogates LPS-delayed neutrophil apoptosis Blood, September 15, 2003; 102(6): 2259 - 2267. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. W. Powell, M. J. Rane, B. A. Joughin, R. Kalmukova, J.-H. Hong, B. Tidor, W. L. Dean, W. M. Pierce, J. B. Klein, M. B. Yaffe, et al. Proteomic Identification of 14-3-3{zeta} as a Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 Substrate: Role in Dimer Formation and Ligand Binding Mol. Cell. Biol., August 1, 2003; 23(15): 5376 - 5387. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Rane, Y. Pan, S. Singh, D. W. Powell, R. Wu, T. Cummins, Q. Chen, K. R. McLeish, and J. B. Klein Heat Shock Protein 27 Controls Apoptosis by Regulating Akt Activation J. Biol. Chem., July 18, 2003; 278(30): 27828 - 27835. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Faderl, D. Harris, Q. Van, H. M. Kantarjian, M. Talpaz, and Z. Estrov Granulocyte-macrophage colony-stimulating factor (GM-CSF) induces antiapoptotic and proapoptotic signals in acute myeloid leukemia Blood, July 15, 2003; 102(2): 630 - 637. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Chen, D. W. Powell, M. J. Rane, S. Singh, W. Butt, J. B. Klein, and K. R. McLeish Akt Phosphorylates p47phox and Mediates Respiratory Burst Activity in Human Neutrophils J. Immunol., May 15, 2003; 170(10): 5302 - 5308. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Park, J. Arcaroli, H.-K. Yum, H. Yang, H. Wang, K.-Y. Yang, K.-H. Choe, D. Strassheim, T. M. Pitts, K. J. Tracey, et al. Activation of gene expression in human neutrophils by high mobility group box 1 protein Am J Physiol Cell Physiol, April 1, 2003; 284(4): C870 - C879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Hasegawa, K. Suzuki, C. Sakamoto, K. Ohta, S. Nishiki, M. Hino, N. Tatsumi, and S. Kitagawa Expression of the inhibitor of apoptosis (IAP) family members in human neutrophils: up-regulation of cIAP2 by granulocyte colony-stimulating factor and overexpression of cIAP2 in chronic neutrophilic leukemia Blood, February 1, 2003; 101(3): 1164 - 1171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. MacKinnon, A. Buckley, E. R. Chilvers, A. G. Rossi, C. Haslett, and T. Sethi Sphingosine Kinase: A Point of Convergence in the Action of Diverse Neutrophil Priming Agents J. Immunol., December 1, 2002; 169(11): 6394 - 6400. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Y. Channon, K. A. Miselis, L. A. Minns, C. Dutta, and L. H. Kasper Toxoplasma gondii Induces Granulocyte Colony-Stimulating Factor and Granulocyte-Macrophage Colony-Stimulating Factor Secretion by Human Fibroblasts: Implications for Neutrophil Apoptosis Infect. Immun., November 1, 2002; 70(11): 6048 - 6057. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Osawa, M. Nagaki, Y. Banno, D. A. Brenner, T. Asano, Y. Nozawa, H. Moriwaki, and S. Nakashima Tumor Necrosis Factor Alpha-Induced Interleukin-8 Production via NF-{kappa}B and Phosphatidylinositol 3-Kinase/Akt Pathways Inhibits Cell Apoptosis in Human Hepatocytes Infect. Immun., November 1, 2002; 70(11): 6294 - 6301. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Saba, G. Soong, S. Greenberg, and A. Prince Bacterial Stimulation of Epithelial G-CSF and GM-CSF Expression Promotes PMN Survival in CF Airways Am. J. Respir. Cell Mol. Biol., November 1, 2002; 27(5): 561 - 567. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Yasui, Y. Sekiguchi, M. Ichikawa, H. Nagumo, T. Yamazaki, A. Komiyama, and H. Suzuki Granulocyte macrophage-colony stimulating factor delays neutrophil apoptosis and primes its function through Ia-type phosphoinositide 3-kinase J. Leukoc. Biol., November 1, 2002; 72(5): 1020 - 1026. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Bozinovski, J. E. Jones, R. Vlahos, J. A. Hamilton, and G. P. Anderson Granulocyte/Macrophage-Colony-stimulating Factor (GM-CSF) Regulates Lung Innate Immunity to Lipopolysaccharide through Akt/Erk Activation of NFkappa B and AP-1 in Vivo J. Biol. Chem., November 1, 2002; 277(45): 42808 - 42814. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Khreiss, L. Jozsef, S. Hossain, J. S. D. Chan, L. A. Potempa, and J. G. Filep Loss of Pentameric Symmetry of C-reactive Protein Is Associated with Delayed Apoptosis of Human Neutrophils J. Biol. Chem., October 18, 2002; 277(43): 40775 - 40781. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Cowburn, K. A. Cadwallader, B. J. Reed, N. Farahi, and E. R. Chilvers Role of PI3-kinase-dependent Bad phosphorylation and altered transcription in cytokine-mediated neutrophil survival Blood, September 18, 2002; 100(7): 2607 - 2616. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-J. Hsu, S.-S. Lee, and W.-W. Lin Polysaccharide purified from Ganoderma lucidum inhibits spontaneous and Fas-mediated apoptosis in human neutrophils through activation of the phosphatidylinositol 3 kinase/Akt signaling pathway J. Leukoc. Biol., July 1, 2002; 72(1): 207 - 216. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Kettritz, M. Choi, W. Butt, M. Rane, S. Rolle, F. C. Luft, and J. B. Klein Phosphatidylinositol 3-Kinase Controls Antineutrophil Cytoplasmic Antibodies--Induced Respiratory Burst in Human Neutrophils J. Am. Soc. Nephrol., July 1, 2002; 13(7): 1740 - 1749. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. E. Kilpatrick, J. Y. Lee, K. M. Haines, D. E. Campbell, K. E. Sullivan, and H. M. Korchak A role for PKC-delta and PI 3-kinase in TNF-alpha -mediated antiapoptotic signaling in the human neutrophil Am J Physiol Cell Physiol, July 1, 2002; 283(1): C48 - C57. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Hoontrakoon, H. W. Chu, S. J. Gardai, S. E. Wenzel, P. McDonald, V. A. Fadok, P. M. Henson, and D. L. Bratton Interleukin-15 Inhibits Spontaneous Apoptosis in Human Eosinophils via Autocrine Production of Granulocyte Macrophage-Colony Stimulating Factor and Nuclear Factor-kappa B Activation Am. J. Respir. Cell Mol. Biol., April 1, 2002; 26(4): 404 - 412. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Kooijman, A. Coppens, and E. Hooghe-Peters IGF-I Inhibits Spontaneous Apoptosis in Human Granulocytes Endocrinology, April 1, 2002; 143(4): 1206 - 1212. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-S. Bae, Y. Kim, J. C. Park, P.-G. Suh, and S. H. Ryu The synthetic chemoattractant peptide, Trp-Lys-Tyr-Met-Val-D-Met, enhances monocyte survival via PKC-dependent Akt activation J. Leukoc. Biol., February 1, 2002; 71(2): 329 - 338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Heyer, S. Saba, R. Adamo, W. Rush, G. Soong, A. Cheung, and A. Prince Staphylococcus aureusagr and sarA Functions Are Required for Invasive Infection but Not Inflammatory Responses in the Lung Infect. Immun., January 1, 2002; 70(1): 127 - 133. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Nijhuis, J.-W. J Lammers, L. Koenderman, and P. J. Coffer Src kinases regulate PKB activation and modulate cytokine and chemoattractant-controlled neutrophil functioning J. Leukoc. Biol., January 1, 2002; 71(1): 115 - 124. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Bouchon, C. Hernandez-Munain, M. Cella, and M. Colonna A Dap12-Mediated Pathway Regulates Expression of Cc Chemokine Receptor 7 and Maturation of Human Dendritic Cells J. Exp. Med., October 15, 2001; 194(8): 1111 - 1122. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Rane, P. Y. Coxon, D. W. Powell, R. Webster, J. B. Klein, W. Pierce, P. Ping, and K. R. McLeish p38 Kinase-dependent MAPKAPK-2 Activation Functions as 3-Phosphoinositide-dependent Kinase-2 for Akt in Human Neutrophils J. Biol. Chem., January 26, 2001; 276(5): 3517 - 3523. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Martin, I. Dransfield, C. Haslett, and A. G. Rossi Cyclic AMP Regulation of Neutrophil Apoptosis Occurs via a Novel Protein Kinase A-independent Signaling Pathway J. Biol. Chem., November 21, 2001; 276(48): 45041 - 45050. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
