A Novel Role of CD40/CD40Ligand Dyad in Regulation of Bone Marrow Granulopoiesis.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4556-4556
Author(s):  
Irene Mavroudi ◽  
Vassiliki Papadaki ◽  
Katerina Pyrovolaki ◽  
Aristides G. Eliopoulos ◽  
Helen A. Papadaki
Keyword(s):  
Tumor Necrosis ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Precursor Cells ◽  
Cell Populations ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Tnfr Superfamily ◽  
Stimulating Factor ◽  
Necrosis Factor

Abstract Abstract 4556 Members of the tumor necrosis factor / tumor necrosis factor receptor (TNF/TNFR) superfamily are involved in bone marrow (BM) homeostasis by regulating the survival, apoptosis, proliferation, and differentiation of hematopoietic progenitor and precursor cells. The CD40 Ligand (CD40L)/CD40 molecules belong to the TNF/TNFR superfamily; we have recently showed that the CD40/CD40L interaction on BM CD34+ cells accelerates the apoptotic cell death. The aim of the present study is to investigate the distribution and function of the CD40/CD40L molecules in the BM granulocytic progenitor and precursor cell populations as well as the effect of their interactions on the stromal release of cytokines related to granulopoiesis. BM samples were obtained from posterior iliac crest aspirates from 19 hematologically healthy subjects after informed consent. We evaluated: (a) the surface expression of CD40 and CD40L on immunomagnetically sorted CD34+, CD34-/CD33+, CD34-/CD33-/CD15+ cells representing sequential stages of the granulocytic development, using flow cytometry in steady state conditions and following 20-hour incubation with 25ng/ml recombinant-human (rh) TNFα; (b) the proportion of apoptotic cells and the level of caspase-3 activation following CD40 engagement in the above cell populations using flow cytometry and a chromo-enzymatic method, respectively; (c) the production of granulocyte-colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) by long-term BM culture (LTBMC) stromal cells upon induction with rhCD40L (1μg/ml), using an enzyme-linked immunoabsorbent assay (ELISA) in culture supernatants. We found that the basal expression of CD40 and CD40L on the CD34+, CD34-/CD33+, CD34-/CD33-/CD15+ cells was 3.34%±3.90% and 1.8%±1.03%, 16.74%±14.69% and 6.96%±4.68%, 3.22%±3.36% and 2.26%±2.71%, respectively. Following incubation with rhTNFα a substantial increase was obtained in the expression of CD40 in all the above cell populations compared to baseline (37.18%±12.51%, 29.25%±14.81%, 15.84%±6.28%, respectively) (p<0.0001, p=0.0003, p=0.0002 respectively). Upon engagement of the TNF-induced CD40 in the CD34+, CD34-/CD33+, CD34-/CD33-/CD15+ cells, a significant increase was obtained in the proportion of apoptotic cells (13.98%±10.43%, 27.24%±19.30%, 25.64%±14.82%, respectively) in comparison to TNF-induced cells without CD40 ligation (5.25%±4.04%, 11.6%±9.04%, 12.42%±5.22%, respectively) (p=0.0071, p=0.0296, p=0.0169, respectively). In keeping with this finding was the significant increase in the caspase-3 activation in all the above populations upon ligation of CD40 compared to baseline (p<0.0001, p=0.0281, p=0.0046, respectively). CD40 was expressed in both CD45+ (8.25%±8.31%) and CD45- (8.46%±4.93%) adherent cell populations (hematopoietic and non hematopoietic, respectively) of confluent LTBMCs (week-3). Following 48-hour incubation of confluent LTBMC stromal layers with rhCD40L, a significant increase was obtained in the production of G-CSF and GM-CSF (2171±2161 pg/ml and 84.10±84.25 pg/ml, respectively) compared to unstimulated cultures (885±1099 pg/ml and 26.57±28.25 pg/ml, respectively) (p=0.0041 and p=0.0074, respectively). These data suggest that the CD40/CD40L interactions, although minimal under steady state conditions, display a dual role in regulation of granulopoiesis under inflammatory conditions by affecting both the BM granylocytic progenitor/precursor cells and the BM microenvironmental cells. More specifically, the expression of CD40 is upregulated in all stages of granulocytic development under the influence of TNFα and upon ligation with CD40L results in acceleration of the apoptotic cell death. The induction of G-CSF and GM-CSF production by BM stroma may represent a compensatory mechanism to rescue granulocytic progenitor/precursor cells from apoptosis. These mechanisms may have a role in the pathogenesis of chronic idiopathic neutropenia where there is evidence for both increased BM TNFα and serum CD40L levels. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Administration of recombinant human granulocyte-macrophage colony- stimulating factor after chemotherapy regulates the expression and secretion of monocyte tumor necrosis factor (TNF) and TNF receptors p55 and p75

Blood ◽  
1995 ◽  
Vol 86 (11) ◽  
pp. 4234-4242 ◽  
Author(s):  
MA Williams ◽  
I Kouroumoussis ◽  
D Syndercombe-Court ◽  
L Hendry ◽  
AC Newland ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Biologically Active ◽  
Colony Stimulating Factor ◽  
Tnf Receptors ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Necrosis Factor

Monocyte expression and secretion of tumor necrosis factor (TNF) and TNF receptors (TNF-R) p55 and p75 was studied in patients receiving granulocyte-macrophage colony-stimulating factor (GM-CSF) after intensive chemotherapy. TNF expression and secretion of biologically active TNF was increased at regeneration compared with that of patients who had received chemotherapy alone. This effect persisted for several weeks after cessation of growth factor therapy. GM-CSF restored the responsiveness of monocytes to bacterial lipopolysaccharide (LPS), which appeared to be diminished after chemotherapy alone. Expression and secretion of TNF-R p55 and p75 by monocytes was augmented by GM-CSF therapy in association with the increase in TNF protein. We propose that GM-CSF administration after chemotherapy restores the normal responsiveness of monocytes to a secondary stimulus such as LPS and primes monocytes to respond to LPS with increased expression and secretion of TNF and TNF-R.

scholarly journals Functional NF-IL6/CCAAT enhancer-binding protein is required for tumor necrosis factor alpha-inducible expression of the granulocyte colony-stimulating factor (CSF), but not the granulocyte/macrophage CSF or interleukin 6 gene in human fibroblasts.

1995 ◽  
Vol 181 (2) ◽  
pp. 793-798 ◽  
Author(s):  
M Kiehntopf ◽  
F Herrmann ◽  
M A Brach
Keyword(s):  
Tumor Necrosis ◽  
Human Fibroblasts ◽  
Transient Transfection ◽  
Tnf Alpha ◽  
Colony Stimulating Factor ◽  
Nf Kappa B ◽  
Gm Csf ◽  
Stimulating Factor ◽  
Necrosis Factor

Tumor necrosis factor (TNF) alpha participates in the regulation of the acute-phase, immune, and inflammatory responses. Target genes known to be transcriptionally activated by TNF-alpha include the granulocyte (G)-colony-stimulating factor (CSF) gene, the granulocyte/macrophage (GM)-CSF gene, as well as the interleukin (IL) 6 gene. Functional nuclear factor (NF)-IL6 recognition sites have been identified in regulatory regions of these genes by transient transfection studies using deleted promoter constructs. In addition, NF-IL6 is known to form heterodimeric complexes with the NF-kappa B transcription factor, which is also engaged in the transcriptional regulation of these genes. The indispensable importance of NF-IL6 for regulating gene expression of proinflammatory cytokine genes in response to inflammatory stimuli in vivo remains, however, unclear. We here report, by using both antisense (AS) oligodesoxyribonucleotide (ODN) and ribozyme (RZ)-mediated specific elimination of NF-IL6 transcripts in human fibroblasts, that TNF-alpha-induced synthesis of G-CSF, but not of GM-CSF or IL-6, is abolished in the absence of functional NF-IL6 in vivo. Both AS ODN and RZ targeting of the NF-IL6 transcript eliminate NF-IL6 protein, as shown in Western blot analysis and electrophoretic mobility shift assays. Similarly, fibroblasts exposed to either the AS NF-IL6 ODN or the NF-IL6 RZ, but not to the sense or nonsense ODN or a mutated ribozyme, also failed to respond with functional activation of NF-IL6 as assayed in transient transfection studies using heterologous promoter constructs harboring the NF-IL6 recognition site. In contrast, protein synthesis, DNA-binding activity, and transcriptional activation capacity of the NF-kappa B transcription factor is not impaired upon exposure to either ODN or RZ. Fibroblasts that had been cultured in the presence of the AS NF-IL6 ODN or the NF-IL6RZ failed to synthesize G-CSF protein in response to TNF-alpha, while TNF-alpha-inducible transcription and release of GM-CSF and IL-6 was preserved.

scholarly journals Mechanisms of tumor necrosis factor-granulocyte-macrophage colony- stimulating factor-induced dendritic cell development

Blood ◽  
1993 ◽  
Vol 82 (10) ◽  
pp. 3019-3028 ◽  
Author(s):  
F Santiago-Schwarz ◽  
N Divaris ◽  
C Kay ◽  
SE Carsons
Keyword(s):  
Dendritic Cell ◽  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Cell Development ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Necrosis Factor

Abstract In a previous report, we described that tumor necrosis factor (TNF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) synergistically enhanced the development of dendritic cell (DC) progeny from early stem cells and that there is a common monocyte-DC progenitor cell. Low levels of DC were obtained with GM-CSF alone, and TNF by itself failed to induce stem cell development. Here, we investigate mechanisms by which TNF and GM-CSF institute increases in DC, and how these same molecules support later stages of DC differentiation. We show that TNF is required as the first signal, that there is upregulation of GM-CSF receptors (GM-CSFRs), and that TNF inhibits the differentiation of colony-forming units-granulocyte. High levels of GM- CSFR were always associated with conditions yielding a large number of DC, and a kinetic analysis showed a close ontogenic relationship between DC and GM-CSFR levels. The addition of anti-GM-CSF or anti-TNF antibodies blocked synergistic responses related to DC development, including high levels of GM-CSFRs. Anti-GM-CSF was the most potent inhibitor of proliferation (80%) and macrophage, DC, and polymorphonuclear (PMN) cell development. With polyclonal anti-TNF, inhibition was less (35%), and there was a shift from myelomonocytic and DC to PMN progeny. Our results support the concept that receptor upregulation is an important mechanism for growth factor synergy. Our data also indicate that the opposing effects of TNF on hematopoiesis contribute to the selection of the DC pathway and emphasize the importance of GM-CSFRs not only in initiated DC development, but also in controlling DC viability and function.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) induces antiapoptotic and proapoptotic signals in acute myeloid leukemia

Blood ◽  
2003 ◽  
Vol 102 (2) ◽  
pp. 630-637 ◽  
Author(s):  
Stefan Faderl ◽  
David Harris ◽  
Quin Van ◽  
Hagop M. Kantarjian ◽  
Moshe Talpaz ◽  
...  
Keyword(s):  
Myeloid Leukemia ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Protein Levels ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Acute Myeloid

AbstractHigh levels of cytokines are associated with a poor prognosis in acute myeloid leukemia (AML). However, cytokines may induce, on one hand, survival factor expression and cell proliferation and, on the other hand, expression of inhibitory signals such as up-regulation of suppressors of cytokine signaling (SOCS) and induce apoptotic cell death. Because blasts from patients with AML express high procaspase protein levels, we asked whether granulocyte-macrophage colony-stimulating factor (GM-CSF) enhances procaspase protein production in AML cells. In the GM-CSF–responsive OCIM2 AML cell line, GM-CSF induced signal transducer and activator of transcription 5 (Stat 5) phosphorylation, up-regulated cyclin D2, and stimulated cell cycle progression. Concurrently, GM-CSF stimulated expression of SOCS-2 and -3 and of procaspases 2 and 3 and induced caspase 3 activation, poly(ADP[adenosine 5′-diphosphate]-ribose) polymerase (PARP) cleavage, and apoptotic cell death. The Janus kinase (Jak)–Stat inhibitor AG490 abrogated GM-CSF–induced expression of procaspase 3 and activation of caspase 3. Under the same conditions GM-CSF up-regulated production of BAX as well as Bcl-2, Bcl-XL, survivin, and XIAP. GM-CSF also increased procaspase 3 protein levels in OCI/AML3 and Mo7e cells, suggesting that this phenomenon is not restricted to a single leukemia cell line. Our data suggest that GM-CSF exerts a dual effect: it stimulates cell division but contemporaneously up-regulates Jak-Stat–dependent proapoptotic proteins. Up-regulation of procaspase levels in AML is thus a beacon for an ongoing growth-stimulatory signal.

scholarly journals Cytokine regulation of colony-stimulating factor production in cultured human synovial fibroblasts: I. Induction of GM-CSF and G-CSF production by interleukin-1 and tumor necrosis factor

Blood ◽  
1990 ◽  
Vol 76 (10) ◽  
pp. 1989-1996 ◽  
Author(s):  
T Leizer ◽  
J Cebon ◽  
JE Layton ◽  
JA Hamilton
Keyword(s):  
Growth Factor ◽  
Tumor Necrosis ◽  
Interleukin 1 ◽  
Synovial Fibroblasts ◽  
Synovial Cells ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Stimulating Factor ◽  
Necrosis Factor ◽  
Csf Production

Abstract The cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF), induce a dose-dependent production of both granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte CSF (G-CSF) in cultured human synovial cells, as measured by immunoassay. With IL-1, significant levels of both CSFs were first detected within 6 to 12 hours, with a maximum reached 24 to 48 hours after commencement of stimulation. A synergistic effect was detected between IL-1 and TNF in production of both CSFs in these cells. No evidence was obtained for the IL-1-induced effect to be mediated by induction of endogenous TNF nor for the TNF-induced stimulation to involve IL-1. IL-1-stimulated synovial cells were shown to secrete biologically active GM-CSF and G- CSF, which were specifically inhibited by their respective monoclonal antibodies. The transcription inhibitor, actinomycin D, and protein synthesis inhibitor, cycloheximide, inhibited the increase in GM-CSF and G-CSF production induced by IL-1 and TNF. Finally, other cytokines, IL-3, interferon gamma (IFN gamma), IL-2, platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and transforming growth factor alpha (TGF alpha), failed to stimulate either GM-CSF or G-CSF production, whether alone or in the presence of IL-1. These results suggest that cytokine-stimulated synovial fibroblasts may be a major source of intraarticular CSF production in the joints of patients with inflammatory arthritis; as a result, monocyte/macrophages and granulocytes may be activated, leading to perpetuation of the inflammation and destructive events occurring in these lesions.

scholarly journals Granulocyte-Macrophage Colony-Stimulating Factor- and Tumor Necrosis Factor Alpha-Mediated Matrix Metalloproteinase Production by Human Osteoblasts and Monocytes after Infection withBrucella abortus

2010 ◽  
Vol 79 (1) ◽  
pp. 192-202 ◽  
Author(s):  
Romina Scian ◽  
Paula Barrionuevo ◽  
Guillermo H. Giambartolomei ◽  
Carlos A. Fossati ◽  
Pablo C. Baldi ◽  
...  
Keyword(s):  
Tumor Necrosis ◽  
Colony Stimulating Factor ◽  
Necrosis Factor Alpha ◽  
Gm Csf ◽  
Tnf Α ◽  
Factor Alpha ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Necrosis Factor

ABSTRACTOsteoarticular complications are common in human brucellosis, but the pathogenic mechanisms involved are largely unknown. Since matrix metalloproteinases (MMPs) are involved in joint and bone damage in inflammatory and infectious diseases, we investigated the production of MMPs by human osteoblasts and monocytes, either uponBrucella abortusinfection or upon reciprocal stimulation with factors produced by each infected cell type.B. abortusinfection of the normal human osteoblastic cell line hFOB 1.19 triggered a significant release of MMP-2, which was mediated in part by granulocyte-macrophage colony-stimulating factor (GM-CSF) acting on these same cells. Supernatants from infected osteoblasts exhibited increased levels of monocyte chemoattractant protein 1 and induced the migration of human monocytes (THP-1 cell line). Infection withB. abortusinduced a high MMP-9 secretion in monocytes, which was also induced by heat-killedB. abortusand by the Omp19 lipoprotein fromB. abortus. These effects were mediated by Toll-like receptor 2 and by the action of tumor necrosis factor alpha (TNF-α) produced by these same cells. Supernatants fromB. abortus-infected monocytes induced MMP-2 secretion in uninfected osteoblasts, and this effect was mediated by TNF-α. Similarly, supernatants from infected osteoblasts induced MMP-9 secretion in uninfected monocytes. This effect was mediated by GM-CSF, which induced TNF-α production by monocytes, which in turn induced MMP-9 in these cells. These results suggest that MMPs could be potentially involved in the tissue damage observed in osteoarticular brucellosis.

scholarly journals Recombinant tumor necrosis factor enhances the proliferative responsiveness of murine peripheral macrophages to macrophage colony- stimulating factor but inhibits their proliferative responsiveness to granulocyte-macrophage colony-stimulating factor

Blood ◽  
1990 ◽  
Vol 75 (8) ◽  
pp. 1627-1632
Author(s):  
BD Chen ◽  
M Mueller
Keyword(s):  
Tumor Necrosis Factor ◽  
Proliferative Response ◽  
Tumor Necrosis ◽  
Colony Stimulating Factor ◽  
Short Term Treatment ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Necrosis Factor

Tumor necrosis factor (TNF) is a protein produced by activated macrophages in response to endotoxin. The effect of recombinant murine TNF (rMuTNF) on the growth of murine tissue-derived macrophage colony- forming units (CFU-M) which are responsive to both macrophage and granulocyte-macrophage colony-stimulating factors (M-CSF and GM-CSF), was studied. TNF alone did not stimulate macrophage proliferation but did prolong their survival in vitro. The proliferative response of CFU- M to M-CSF, however, was greatly enhanced by the presence of TNF. The enhancement effect of TNF is dose-dependent, reaching a maximum at approximately 50 U/mL. In contrast, the proliferative responsiveness of CFU-M to GM-CSF was inhibited by the concurrent addition of rMuTNF. Both effects appear to be caused directly by rMuTNF, rather than by the secondary factor(s) produced by TNF-treated macrophages. TNF treatment also induced a transient downmodulation of M-CSF receptors in cultured macrophages and accelerated their uptake and use of exogenous M-CSF, which may account for, at least in part, the enhanced proliferative activity in response to M-CSF. Short-term treatment (24 hours) was not sufficient to induce either an enhancing or an inhibitory effect upon CFU-M. This study suggests an autoregulatory role for TNF in the production of mature tissue macrophages by selectively enhancing their proliferative response to lineage specific growth factor, M-CSF.

scholarly journals Recombinant human TNF alpha stimulates production of granulocyte colony- stimulating factor

Blood ◽  
1987 ◽  
Vol 70 (1) ◽  
pp. 55-59 ◽  
Author(s):  
HP Koeffler ◽  
J Gasson ◽  
J Ranyard ◽  
L Souza ◽  
M Shepard ◽  
...  
Keyword(s):  
Tumor Necrosis ◽  
Tnf Alpha ◽  
Fibroblast Cells ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Necrosis Factor Alpha ◽  
Gm Csf ◽  
Factor Alpha ◽  
Stimulating Factor ◽  
Necrosis Factor

Tumor necrosis factor alpha (TNF alpha) stimulates production of granulocyte colony-stimulating factor (G-CSF) protein and mRNA in fibroblast cells. In contrast, TNF beta is at least 1,000-fold less potent than TNF alpha in increasing levels of G-CSF and GM-CSF activity and mRNA. The fibroblasts produce G-CSF and GM-CSF mRNA in a coordinate fashion after exposure to TNF. The ability of TNF alpha to stimulate production of CSF in fibroblasts may be pivotal to regulation of hematopoiesis.

scholarly journals Production of interleukin-1 alpha, interleukin-1 beta and tumor necrosis factor by human mononuclear cells stimulated with granulocyte- macrophage colony-stimulating factor [see comments]

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1368-1374 ◽  
Author(s):  
SD Sisson ◽  
CA Dinarello
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Mononuclear Cells ◽  
Interleukin 1 ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Necrosis Factor

Abstract Recent studies have examined the synergistic effects of granulocyte- macrophage colony-stimulating factor (GM-CSF) and hematopoietin-1 (now identified as Interleukin-1, IL-1) on bone marrow colony formation. In the present report, human peripheral blood mononuclear cells (MNCs) were stimulated in vitro with recombinant human GM-CSF (rGM-CSF) and production of IL-1 alpha, IL-1 beta, and tumor necrosis factor (TNF) was measured by specific radioimmunoassays. In the MNCs of 20 individuals, rGM-CSF's ability to induce the three cytokines was variable. Nearly all donors responded to low-dose rGM-CSF (0.02 to 2 ng/mL) with production of TNF, whereas some individuals did not produce IL-1 alpha or IL-1 beta. The MNCs from some subjects stimulated with high-dose rGM-CSF (10 to 80 ng/mL) produced as much cytokine as in response to 10 ng/mL endotoxin. Localization (ie, extracellular or cell- associated cytokine) was specific for the cytokine rather than the stimulus. Indomethacin increased the amount of cytokine produced in response to rGM-CSF for IL-1 beta and TNF but not for IL-1 alpha. In addition, interferon-gamma (INF-gamma) upregulated the amount of TNF induced by rGM-CSF in all donors examined, with variable effect on IL-1 alpha and IL-1 beta. Suboptimal levels of endotoxin incubated with rGM- CSF did not alter the amount of IL-1 produced as compared with cells stimulated with rGM-CSF alone, whereas TNF production showed either no change or a slight decrease in production. These data suggest that GM- CSF may play an important role in the host defense response by stimulating production of these cytokines.

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