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1.
Mucosal Immunol ; 9(6): 1372-1383, 2016 11.
Article in English | MEDLINE | ID: mdl-26906405

ABSTRACT

Innate lymphoid cells (ILCs) have a central role in innate defenses against pathogens, lymphoid organogenesis, and tissue remodeling. They have been detected in human decidua, however, their role in this tissue remains unclear. Successful pregnancy requires an early inflammatory phase favoring implantation and tissue remodeling as well as a subsequent regulatory phase to prevent fetal rejection and supporting neoangiogenesis. Here, we show that, during the first trimester of pregnancy, neutrophils infiltrate decidua basalis and are more abundant in normal pregnancy than in spontaneous miscarriages. Decidual neutrophils localize in proximity of NCR+ILC3, which may influence neutrophil migration and survival given their production of CXCL8 and granulocyte macrophage colony-stimulating factor (GM-CSF). Moreover, NCR+ILC3-derived GM-CSF was found to induce the expression of heparin-binding EGF-like growth factor and IL1ra in neutrophils, two proteins/cytokines involved in tissue remodeling and maintenance of pregnancy. Our data suggest that the simultaneous presence of NCR+ILC3 and neutrophils in decidual tissues and their possible cross talk, may have a role in the early phases of pregnancy.


Subject(s)
Chemotaxis, Leukocyte/immunology , Decidua/immunology , Decidua/metabolism , Immunity, Innate , Lymphocyte Subsets/immunology , Lymphocyte Subsets/metabolism , Neutrophils/immunology , Neutrophils/metabolism , Antigens, CD/metabolism , Biomarkers , Cell Survival/immunology , Cytokines/metabolism , Female , Gene Expression , Heparin-binding EGF-like Growth Factor/biosynthesis , Heparin-binding EGF-like Growth Factor/genetics , Humans , Lymphocyte Activation/immunology , Neutrophil Infiltration/immunology , Phenotype , Pregnancy , Pregnancy Trimester, First
2.
Clin Mol Allergy ; 13(1): 16, 2015.
Article in English | MEDLINE | ID: mdl-26240517

ABSTRACT

BACKGROUND: TNF-like ligand 1A (TL1A), a recently recognized member of the TNF superfamily, and its death domain receptor 3 (DR3), firstly identified for their relevant role in T lymphocyte homeostasis, are now well-known mediators of several immune-inflammatory diseases, ranging from rheumatoid arthritis to inflammatory bowel diseases to psoriasis, whereas no data are available on their involvement in sarcoidosis, a multisystemic granulomatous disease where a deregulated T helper (Th)1/Th17 response takes place. METHODS: In this study, by flow cytometry, real-time PCR, confocal microscopy and immunohistochemistry analyses, TL1A and DR3 were investigated in the pulmonary cells and the peripheral blood of 43 patients affected by sarcoidosis in different phases of the disease (29 patients with active sarcoidosis, 14 with the inactive form) and in 8 control subjects. RESULTS: Our results demonstrated a significant higher expression, both at protein and mRNA levels, of TL1A and DR3 in pulmonary T cells and alveolar macrophages of patients with active sarcoidosis as compared to patients with the inactive form of the disease and to controls. In patients with sarcoidosis TL1A was strongly more expressed in the lung than the blood, i.e., at the site of the involved organ. Additionally, zymography assays showed that TL1A is able to increase the production of matrix metalloproteinase 9 by sarcoid alveolar macrophages characterized, in patients with the active form of the disease, by reduced mRNA levels of the tissue inhibitor of metalloproteinase (TIMP)-1. CONCLUSIONS: These data suggest that TL1A/DR3 interactions are part of the extended and complex immune-inflammatory network that characterizes sarcoidosis during its active phase and may contribute to the pathogenesis and to the progression of the disease.

4.
Clin Exp Immunol ; 178(1): 129-41, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24853271

ABSTRACT

The role of dendritic cells (DCs) and macrophages in allogeneic haematopoietic stem cell transplant (HSCT) is critical in determining the extent of graft-versus-host response. The goal of this study was to analyse slanDCs, a subset of human proinflammatory DCs, in haematopoietic stem cell (HSC) sources, as well as to evaluate their 1-year kinetics of reconstitution, origin and functional capacities in peripheral blood (PB) and bone marrow (BM) of patients who have undergone HSCT, and their presence in graft-versus-host disease (GVHD) tissue specimens. slanDCs were also compared to myeloid (m)DCs, plasmacytoid (p)DCs and monocytes in HSC sources and in patients' PB and BM throughout reconstitution. slanDCs accounted for all HSC sources. In patients' PB and BM, slanDCs were identified from day +21, showing median frequencies comparable to healthy donors, donor origin and kinetics of recovery similar to mDCs, pDCs, and monocytes. Under cyclosporin treatment, slanDCs displayed a normal pattern of maturation, and maintained an efficient chemotactic activity and capacity of releasing tumour necrosis factor (TNF)-α upon lipopolysaccharide (LPS) stimulation. None the less, they were almost undetectable in GVHD tissue specimens, being present only in intestinal acute GVHD samples. slanDCs reconstitute early, being donor-derived and functionally competent. The absence of slanDCs from most of the GVHD-targeted tissue specimens seems to rule out the direct participation of these cells in the majority of the local reactions characterizing GVHD.


Subject(s)
Dendritic Cells/immunology , Hematopoietic Stem Cells/immunology , Adult , Female , Graft vs Host Disease/immunology , Hematopoietic Stem Cell Transplantation/methods , Humans , Male , Middle Aged , Monocytes/immunology , Tissue Donors , Transplantation, Homologous/methods , Tumor Necrosis Factor-alpha/immunology , Young Adult
5.
Br J Haematol ; 137(6): 553-9, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17539776

ABSTRACT

B-lymphocyte stimulator (BLyS) acts as survival factor for B lymphocytes. As Hodgkin and Reed-Sternberg (HRS) cells express receptors through which BLyS promotes their growth and chemotherapy resistance, we investgated whether this molecule was increased in sera from patients with classical Hodgkin lymphoma (cHL) and whether it correlates with clinical-pathological features and outcomes. Enzyme-linked immunosorbent assay was used to measure soluble BLyS (sBLyS) in sera from 87 patients and 33 donors; higher levels were detected in patients (mean +/- standard error 4493.9 +/- 264.9 pg/ml vs. 2687.0 +/- 200.9 pg/ml; P < 0.0001). Levels above the median value (4242.0 pg/ml) were associated with age > or = 45 years (P = 0.042), advanced stages of disease (P = 0.005), systemic symptoms (P = 0.014) and extranodal involvement (P = 0.009). Five-year failure-free survival (FFS) of patients with sBLyS below or equal to median levels was 88.6% as compared to 65.1% of those with levels above the median (P = 0.009). Statistical analyses confirmed the prognostic significance of sBLyS (P = 0.046). When patients were analysed according to variables associated with high levels, sBLyS showed an independent predictive power in terms of FFS. Our findings support the involvement of BLyS in cHL pathogenesis. The association between high serum levels and an inferior FFS indicates that sBLyS is a possible prognostic predictor with a potential significance as a therapeutic target.


Subject(s)
B-Cell Activating Factor/blood , Biomarkers, Tumor/blood , Hodgkin Disease/blood , Adolescent , Adult , Aged , Case-Control Studies , Enzyme-Linked Immunosorbent Assay/methods , Female , Hodgkin Disease/mortality , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Predictive Value of Tests , Prognosis , Treatment Outcome
6.
J Immunol ; 167(4): 2312-22, 2001 Aug 15.
Article in English | MEDLINE | ID: mdl-11490020

ABSTRACT

We have recently shown that IL-10 fails to trigger Stat3 and Stat1 tyrosine phosphorylation in freshly isolated human neutrophils. In this study, we report that IL-10 can nonetheless induce Stat3 tyrosine phosphorylation and the binding of Stat1 and Stat3 to the IFN-gamma response region or the high-affinity synthetic derivative of the c-sis-inducible element in neutrophils that have been cultured for at least 3 h with LPS. Similarly, the ability of IL-10 to up-regulate suppressor of cytokine signaling (SOCS)-3 mRNA was dramatically enhanced in cultured neutrophils and, as a result, translated into the SOCS-3 protein. Since neutrophils' acquisition of responsiveness to IL-10 required de novo protein synthesis, we assessed whether expression of IL-10R1 or IL-10R2 was modulated in cultured neutrophils. We detected constitutive IL-10R1 mRNA and protein expression in circulating neutrophils, at levels which were much lower than those observed in autologous monocytes or lymphocytes. In contrast, IL-10R2 expression was comparable in both cell types. However, IL-10R1 (but not IL-10R2) mRNA and protein expression was substantially increased in neutrophils stimulated by LPS. The ability of IL-10 to activate Stat3 tyrosine phosphorylation and SOCS-3 synthesis and to regulate IL-1 receptor antagonist and macrophage-inflammatory protein 1beta release in LPS-treated neutrophils correlated with this increased IL-10R1 expression, and was abolished by neutralizing anti-IL-10R1 and anti-IL-10R2 Abs. Our results demonstrate that the capacity of neutrophils to respond to IL-10, as assessed by Stat3 tyrosine phosphorylation, SOCS-3 expression, and modulation of cytokine production, is very dependent on the level of expression of IL-10R1.


Subject(s)
Interleukin-10/metabolism , Neutrophils/immunology , Neutrophils/metabolism , Receptors, Interleukin/biosynthesis , Repressor Proteins , Transcription Factors , Up-Regulation/immunology , Cell Separation , Cells, Cultured , Cytokines/metabolism , DNA-Binding Proteins/metabolism , Humans , Immune Sera/pharmacology , Interleukin-10/pharmacology , Interleukin-10/physiology , Lipopolysaccharides/pharmacology , Phosphorylation , Protein Binding/immunology , Protein Biosynthesis , Proteins/genetics , Proteins/physiology , RNA, Messenger/biosynthesis , Receptors, Interleukin/antagonists & inhibitors , Receptors, Interleukin/genetics , Receptors, Interleukin/immunology , Receptors, Interleukin-10 , STAT3 Transcription Factor , Signal Transduction/genetics , Signal Transduction/immunology , Suppressor of Cytokine Signaling 3 Protein , Suppressor of Cytokine Signaling Proteins , Trans-Activators/metabolism , Tyrosine/metabolism
7.
Eur J Immunol ; 31(7): 1981-8, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11449350

ABSTRACT

Macrophage inflammatory protein-3alpha (MIP-3alpha)/CCL20 and MIP-3beta/CCL19 are members of the CC chemokine subfamily which exert their effects through specific receptors, CCR6 and CCR7, respectively. Previously, we have reported that human neutrophils have the capacity to produce a number of chemokines, including IL-8/CXCL8, GROalpha/CXCL1, IP-10/CXCL10, and MIG/CXCL9. Herein, we show that neutrophils also have the ability to express and release MIP-3alpha/CCL20 and MIP-3beta/CCL19 when cultured with either LPS or TNF-alpha. We also report that MIP-3alpha/CCL20 and MIP-3beta/CCL19 production by LPS-stimulated neutrophils is negatively modulated by IL-10. Remarkably, we found that supernatants harvested from stimulated neutrophils not only induced chemotaxis of both immature and mature dendritic cells (DC), but also triggered rapid integrin-dependent adhesion of CCR6- and CCR7-expressing lymphocytes to purified VCAM-1 and ICAM-1, respectively. Importantly, both chemotaxis and rapid integrin-dependent adhesion were dramatically suppressed by anti-MIP-3alpha/CCL20 and anti-MIP-3beta/CCL19 neutralizing antibodies, indicating that MIP-3alpha/CCL20 and MIP-3beta/CCL19 present in the supernatants were both biologically active. As these chemokines are primarily chemotactic for DC and specific lymphocyte subsets, the ability of neutrophils to produce MIP-3alpha/CCL20 and MIP-3beta/CCL19 might be significant in orchestrating the recruitment of these cell types to the inflamed sites and therefore in contributing to the regulation of the immune response.


Subject(s)
Chemokines, CC/biosynthesis , Macrophage Inflammatory Proteins/biosynthesis , Neutrophils/immunology , Receptors, Chemokine , Animals , Cell Adhesion , Cell Line , Cells, Cultured , Chemokine CCL19 , Chemokine CCL20 , Chemokines, CC/genetics , Chemotaxis , Culture Media, Conditioned , Dendritic Cells/immunology , Humans , Intercellular Adhesion Molecule-1/physiology , Lipopolysaccharides/pharmacology , Macrophage Inflammatory Proteins/genetics , Mice , Mice, Inbred BALB C , Neutrophils/drug effects , RNA, Messenger/biosynthesis , Receptors, CCR6 , Tumor Necrosis Factor-alpha/pharmacology , Vascular Cell Adhesion Molecule-1/physiology
8.
Infect Immun ; 68(12): 6917-23, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11083814

ABSTRACT

Accumulation of polymorphonuclear neutrophils (PMN) into the subarachnoidal space is one of the hallmarks of Neisseria meningitidis infection. In this study, we evaluated the ability of outer membrane vesicles (OMV) from N. meningitidis B to stimulate cytokine production by neutrophils. We found that PMN stimulated in vitro by OMV produce proinflammatory cytokines and chemokines including tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-8, macrophage inflammatory protein 1alpha (MIP-1alpha), and MIP-1beta. A considerable induction of gamma interferon (IFN-gamma)-inducible protein 10 (IP-10) mRNA transcripts, as well as extracellular IP-10 release, was also observed when neutrophils were stimulated by OMV in combination with IFN-gamma. Furthermore, PMN stimulated by OMV in the presence of IFN-gamma demonstrated an enhanced capacity to release TNF-alpha, IL-1beta, IL-8, and MIP-1beta compared to stimulation with OMV alone. In line with its downregulatory effects on neutrophil-derived proinflammatory cytokines, IL-10 potently inhibited TNF-alpha, IL-1beta, IL-8, and MIP-1beta production triggered by OMV. Finally, a neutralizing anti-TNF-alpha monoclonal antibody (MAb) did not influence the release of IL-8 and MIP-1beta induced by OMV, therefore excluding a role for endogenous TNF-alpha in mediating the induction of chemokine release by OMV. In contrast, the ability of lipopolysaccharide from N. meningitidis B to induce the production of IL-8 and MIP-1beta was significantly inhibited by anti-TNF-alpha MAb. Our results establish that, in response to OMV, neutrophils produce a proinflammatory profile of cytokines and chemokines which may not only play a role in the pathogenesis of meningitis but may also contribute to the development of protective immunity to serogroup B meningococci.


Subject(s)
Chemokines, CXC/biosynthesis , Interleukin-1/biosynthesis , Interleukin-8/biosynthesis , Macrophage Inflammatory Proteins/biosynthesis , Neisseria meningitidis/immunology , Neutrophils/metabolism , Tumor Necrosis Factor-alpha/biosynthesis , Chemokine CCL3 , Chemokine CCL4 , Chemokine CXCL10 , Chemokines, CXC/genetics , Humans , Interferon-gamma/pharmacology , Interleukin-1/genetics , Interleukin-10/pharmacology , Interleukin-8/genetics , Macrophage Inflammatory Proteins/genetics , Meningitis, Meningococcal/etiology , RNA, Messenger/analysis , Tumor Necrosis Factor-alpha/genetics
9.
Curr Opin Hematol ; 7(3): 174-7, 2000 May.
Article in English | MEDLINE | ID: mdl-10786655

ABSTRACT

Interleukin-15 is a recently discovered cytokine produced by several cell types (including fibroblasts, keratinocytes, endothelial cells, and macrophages) in response to endotoxin or microbial infection. In turn, interleukin-15 has been shown to act on various cells of the immune system, including T and B lymphocytes, natural killer cells, monocytes, eosinophils, and circulating neutrophils. In the latter instance, interleukin-15 was initially observed to induce cytoskeletal rearrangements, to enhance phagocytosis, to increase the synthesis of several cellular proteins, and to delay apoptosis. Recently, interleukin-15 has been found to elicit other functional responses in neutrophils, such as chemokine production. This review recapitulates advances made in the area of interleukin-15/neutrophil interactions.


Subject(s)
Interleukin-15/pharmacology , Neutrophils/drug effects , Animals , Humans , Inflammation , Interleukin-15/blood , Interleukin-15/physiology , Neutrophil Activation/drug effects , Neutrophil Activation/genetics , Neutrophils/cytology , Neutrophils/physiology
10.
Immunol Rev ; 177: 195-203, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11138776

ABSTRACT

Neutrophils are known to play an important role in inflammatory responses by virtue of their ability to perform a series of effector functions that collectively represent a major mechanism of innate immunity against injury and infection. In recent years, however, it has become obvious that the contribution of neutrophils to host defence and natural immunity extends well beyond their traditional role as professional phagocytes. Indeed, neutrophils can be induced to express a number of genes whose products lie at the core of inflammatory and immune responses. These include not only Fc receptors, complement components, cationic antimicrobial and NADPH oxidase proteins, but also a variety of cytokines (including tumour necrosis factor-alpha, interleukin (IL)-1beta, IL-1R alpha, IL-12 and vascular endothelial growth factor), and chemokines such as IL-8, growth-related gene product, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, interferon-gamma-inducible protein of 10 kDa and monokine induced by interferon-gamma. Because these chemokines are primarily chemotactic for neutrophils, monocytes, immature dendritic cells and T-lymphocyte subsets, a potential role for neutrophils in orchestrating the sequential recruitment of distinct leukocyte types to the inflamed tissue is likely to occur. The purpose of this review is to summarize the essential features of the production of chemokines by polymorphonuclear neutrophil leukocytes and the contribution that we have made to characterize some aspects of this newly discovered crucial function of neutrophils.


Subject(s)
Chemokines/immunology , Neutrophils/immunology , Animals , Humans , Inflammation , Neutrophil Activation
11.
Blood ; 94(9): 3169-77, 1999 Nov 01.
Article in English | MEDLINE | ID: mdl-10556204

ABSTRACT

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a widely expressed EGF superfamily member that induces mitogenic and/or chemotactic activities toward different cell types through binding to EGF receptors 1 or 4. Membrane-bound HB-EGF exerts growth activity and adhesion capabilities and possesses the unique property of being the receptor for diphtheria toxin (DT). Using molecular and functional techniques, we show that human polymorphonuclear granulocytes (PMN), which did not express HB-EGF in resting conditions, expressed it at mRNA and protein level, following incubation with granulocyte-macrophage colony-stimulating factor (GM-CSF). Other classic agonists for PMN (including lipopolysaccharide, phagocytable particles, tumor necrosis factor-alpha, or G-CSF) failed to induce HB-EGF. The effects of GM-CSF on HB-EGF mRNA levels were concentration-dependent, reached a plateau after 1 to 2 hours of stimulation, and did not require protein synthesis. After GM-CSF treatment, membrane-bound HB-EGF was detected by flow cytometry. At the same time, PMN acquired sensitivity to the apoptosis-promoting effect of DT, which, moreover, specifically suppressed the GM-CSF-induced priming of formyl-methionyl-leucyl-phenylalanine-stimulated superoxide anion release. Finally, soluble HB-EGF was detected in the PMN culture medium by a specific enzyme-linked immunosorbent assay. Thus, we provide evidence that HB-EGF is specifically inducible by GM-CSF in PMN and represents a novel peptide to be included in the repertoire of PMN-derived cytokines.


Subject(s)
Diphtheria Toxin/pharmacology , Epidermal Growth Factor/biosynthesis , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Neutrophils/metabolism , Receptors, Cell Surface/biosynthesis , Flow Cytometry , Heparin-binding EGF-like Growth Factor , Humans , Intercellular Signaling Peptides and Proteins , RNA, Messenger/biosynthesis
12.
Blood ; 94(8): 2880-9, 1999 Oct 15.
Article in English | MEDLINE | ID: mdl-10515892

ABSTRACT

We have recently shown that, in human neutrophils, interleukin-10 (IL-10) fails to induce specific DNA-binding activities to the gamma-interferon response region (GRR), a regulatory element located in the FcgammaRI gene promoter, which is required for transcriptional activation by IL-10 and interferon gamma (IFNgamma) in monocytic cells. In this study, we report that IL-10 is also unable to induce the binding of STAT1 or STAT3 to the serum-inducible element (hSIE/m67), despite the fact that both proteins are expressed in neutrophils. Whereas IFNgamma and granulocyte colony-stimulating factor (G-CSF) are efficient inducers of STAT1 and STAT3 tyrosine phosphorylation in polymorphonuclear neutrophils (PMN), IL-10 fails to trigger STAT1 and STAT3 tyrosine and serine phosphorylation, therefore explaining its inability to induce the FcgammaRI expression in these cells. By contrast, we demonstrate that IL-10 alone represents an efficient stimulus of CIS3/SOCS3 mRNA expression in neutrophils. CIS3/SOCS3 belongs to the recently cloned cytokine-inducible SH2-containing protein (CIS) gene family (which also includes CIS1, CIS2, CIS4, CIS5, and JAB) that is believed to be, at least in part, under the control of STAT transcription factors and whose products are potential modulators of cytokine signaling. Moreover, IL-10 synergizes with lipopolysaccharide (LPS) in upregulating CIS3/SOCS3 mRNA expression in PMN through a mechanism that involves mRNA stabilization. In contrast to CIS3/SOCS3, mRNA transcripts encoding other family members are unaffected by IL-10 in neutrophils. Finally, transfection of CIS3/SOCS3 in murine M1 myeloid cells suppresses LPS-induced growth arrest, macrophage-like differentiation, and nitric oxide synthesis, but not IL-6 mRNA expression. Collectively, our data suggest that, in neutrophils, the activation of STAT1 and STAT3 phosphorylation is neither required for CIS3/SOCS3 induction by IL-10 nor involved in the regulatory effects of IL-10 on cytokine production.


Subject(s)
Gene Expression Regulation/drug effects , Interleukin-10/pharmacology , Neutrophils/drug effects , Protein Biosynthesis , RNA, Messenger/biosynthesis , Repressor Proteins , Signal Transduction/drug effects , Transcription Factors , Animals , Cell Differentiation/drug effects , Cells, Cultured , DNA-Binding Proteins/metabolism , Enzyme Activation/drug effects , Granulocyte Colony-Stimulating Factor/pharmacology , Humans , Interferon-gamma/pharmacology , Lipopolysaccharides/pharmacology , Mice , Multigene Family , Neutrophils/metabolism , Nitric Oxide/biosynthesis , Phosphorylation , Protein Processing, Post-Translational/drug effects , Proteins/genetics , RNA, Messenger/genetics , STAT1 Transcription Factor , STAT3 Transcription Factor , Suppressor of Cytokine Signaling 3 Protein , Suppressor of Cytokine Signaling Proteins , Tetradecanoylphorbol Acetate/pharmacology , Trans-Activators/metabolism , Transfection , src Homology Domains
14.
Int J Clin Lab Res ; 29(1): 41-5, 1999.
Article in English | MEDLINE | ID: mdl-10356663

ABSTRACT

In this study, we investigated the expression of Bak, a member of the Bcl-2 protein family, in human polymorphonuclear neutrophils. Northern blot and Western blot analyses revealed that Bak messenger RNA and protein were constitutively expressed in peripheral polymorphonuclear neutrophils and mononuclear cells, as well as in several hematopoietic cell lines. Remarkably, culturing neutrophils for 24 h in the presence or absence of interferon-gamma or tumor necrosis factor-alpha, which have been described to modulate the survival rate of these cells, did not influence the expression of antigenic Bak. Taken together, our data indicate that the expression of the pro-apoptotic protein Bak in polymorphonuclear neutrophils is constitutive, is not subject to modulation, and does not correlate with the neutrophil life span in culture.


Subject(s)
Apoptosis/immunology , Membrane Proteins/analysis , Neutrophils/chemistry , Neutrophils/immunology , Antibodies , Blotting, Western , Cell Survival/drug effects , Cell Survival/immunology , Cells, Cultured , Humans , Interferon-gamma/pharmacology , Membrane Proteins/immunology , Neutrophils/cytology , Tumor Necrosis Factor-alpha/pharmacology , bcl-2 Homologous Antagonist-Killer Protein
15.
J Neuroimmunol ; 93(1-2): 45-52, 1999 Jan 01.
Article in English | MEDLINE | ID: mdl-10378868

ABSTRACT

Growing evidence indicates that amyloid (A beta) deposition and phagocyte activation participate in inflammatory reactions in the brain during the course of Alzheimer's disease. To further investigate the effects of A beta-phagocyte interaction, we examined the production of proinflammatory (IL-1beta, IL-6), chemotactic (MIP-1alpha, IP-10) and inhibitory (IL-1Ra, IL-10 and TGFbeta1) cytokines by cultured human monocytes and mouse microglial cells upon stimulation with A beta[25-35]. Northern blot analysis and specific immunoassays demonstrated that A beta[25-35] triggers mRNA expression and release of IL-1beta, IL-1Ra and MIP-1alpha but not of IL-6, IL-10, TGFbeta1 and IP-10 from human monocytes. Similar results were obtained by examining the production of IL-1beta, IL-6 and IL-10 from mouse microglial cells in the same experimental conditions. Taken together, these data indicate that A beta-phagocyte interaction can drive a different response towards cytokine production by monocytes and microglia, with a particular proinflammatory trend, and further support a role for A beta deposition as a triggering factor of inflammatory events in Alzheimer's disease.


Subject(s)
Amyloid beta-Peptides/pharmacology , Microglia/immunology , Monocytes/immunology , Peptide Fragments/pharmacology , Alzheimer Disease/immunology , Animals , Antineoplastic Agents/immunology , Antirheumatic Agents/immunology , Cell Line , Chemokine CCL3 , Chemokine CCL4 , Chemokine CXCL10 , Chemokines, CXC/immunology , Gene Expression Regulation/immunology , Humans , Interleukin 1 Receptor Antagonist Protein , Interleukin-1/genetics , Interleukin-1/immunology , Interleukin-10/genetics , Interleukin-10/immunology , Interleukin-6/genetics , Interleukin-6/immunology , Macrophage Inflammatory Proteins/genetics , Macrophage Inflammatory Proteins/immunology , Mice , Microglia/cytology , Microglia/drug effects , Monocytes/cytology , Monocytes/drug effects , Neuritis/immunology , Plaque, Amyloid/immunology , RNA, Messenger/metabolism , Sialoglycoproteins/genetics , Sialoglycoproteins/immunology , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/immunology
16.
J Immunol ; 162(8): 4928-37, 1999 Apr 15.
Article in English | MEDLINE | ID: mdl-10202039

ABSTRACT

Monokine induced by IFN-gamma (MIG), IFN-inducible T cell alpha chemoattractant (I-TAC), and IFN-gamma-inducible protein of 10 kDa (IP-10) are related members of the CXC chemokine subfamily that bind to a common receptor, CXCR3, and that are produced by different cell types in response to IFN-gamma. We have recently reported that human polymorphonuclear neutrophils (PMN) have the capacity to release IP-10. Herein, we show that PMN also have the ability to produce MIG and to express I-TAC mRNA in response to IFN-gamma in combination with either TNF-alpha or LPS. While IFN-gamma, alone or in association with agonists such as fMLP, IL-8, granulocyte (G)-CSF and granulocyte-macrophage (GM)-CSF, failed to influence MIG, IP-10, and I-TAC gene expression, IFN-alpha, in combination with TNF-alpha, LPS, or IL-1beta, resulted in a considerable induction of IP-10 release by neutrophils. Furthermore, IL-10 and IL-4 significantly suppressed the expression of MIG, IP-10, and I-TAC mRNA and the extracellular production of MIG and IP-10 in neutrophils stimulated with IFN-gamma plus either LPS or TNF-alpha. Finally, supernatants harvested from stimulated PMN induced migration and rapid integrin-dependent adhesion of CXCR3-expressing lymphocytes; these activities were significantly reduced by neutralizing anti-MIG and anti-IP-10 Abs, suggesting that they were mediated by MIG and IP-10 present in the supernatants. Since MIG, IP-10, and I-TAC are potent chemoattractants for NK cells and Th1 lymphocytes, the ability of neutrophils to produce these chemokines might contribute not only to the progression and evolution of the inflammatory response, but also to the regulation of the immune response.


Subject(s)
Chemokines, CXC/genetics , Gene Expression Regulation/immunology , Intercellular Signaling Peptides and Proteins , Interferon-gamma/pharmacology , Neutrophils/metabolism , Apoptosis/drug effects , Apoptosis/immunology , Cell Adhesion/immunology , Cell-Free System/chemistry , Cell-Free System/immunology , Cells, Cultured , Chemokine CXCL10 , Chemokine CXCL11 , Chemokine CXCL9 , Chemokines, CXC/biosynthesis , Chemokines, CXC/metabolism , Chemotaxis, Leukocyte/immunology , Eosinophils/metabolism , Humans , Interferon-alpha/pharmacology , Interleukin-10/pharmacology , Interleukin-4/pharmacology , Leukocytes, Mononuclear/metabolism , Lipopolysaccharides/pharmacology , Neutrophil Activation , Neutrophils/cytology , Neutrophils/drug effects , RNA, Messenger/biosynthesis , Tumor Necrosis Factor-alpha/pharmacology
17.
18.
Blood ; 93(5): 1715-23, 1999 Mar 01.
Article in English | MEDLINE | ID: mdl-10029601

ABSTRACT

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is an EGF family member expressed by numerous cell types that binds to EGF receptor 1 (HER-1) or 4 (HER-4) inducing mitogenic and/or chemotactic activities. Membrane-bound HB-EGF retains growth activity and adhesion capabilities and the unique property of being the receptor for diphtheria toxin (DT). The interest in studying HB-EGF in acute leukemia stems from these mitogenic, chemotactic, and receptor functions. We analyzed the expression of HB-EGF in L428, Raji, Jurkat, Karpas 299, L540, 2C8, HL-60, U937, THP-1, ML-3, and K562 cell lines and in primary blasts from 12 acute myeloid leukemia (AML) cases, by reverse-transcriptase polymerase chain reaction (RT-PCR) and Northern blot and by the evaluation of sensitivity to DT. The release of functional HB-EGF was assessed by evaluation of its proliferative effects on the HB-EGF-sensitive Balb/c 3T3 cell line. HB-EGF was expressed by all myeloid and T, but not B (L428, Raji), lymphoid cell lines tested, as well as by the majority (8 of 12) of ex vivo AML blasts. Cell lines (except for the K562 cell line) and AML blasts expressing HB-EGF mRNA underwent apoptotic death following exposure to DT, thus demonstrating the presence of the HB-EGF molecule on their membrane. Leukemic cells also released a fully functional HB-EGF molecule that was mitogenic for the Balb/c 3T3 cell line. Factors relevant to the biology of leukemic growth, such as tumor necrosis factor-alpha (TNF-alpha), 1alpha,25-(OH)2D3, and especially all-trans retinoic acid (ATRA), upregulated HB-EGF mRNA in HL-60 or ML-3 cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) induced HB-EGF mRNA and acquisition of sensitivity to DT in one previously HB-EGF-negative leukemia case. Moreover, the U937 and Karpas 299 cell lines expressed HER-4 mRNA. This work shows that HB-EGF is a growth factor produced by primary leukemic cells and regulated by ATRA, 1alpha, 25-(OH)2D3, and GM-CSF.


Subject(s)
Epidermal Growth Factor/biosynthesis , Leukemia, Myeloid/metabolism , Acute Disease , Adult , Aged , Apoptosis , Female , HL-60 Cells , Heparin-binding EGF-like Growth Factor , Humans , Intercellular Signaling Peptides and Proteins , Jurkat Cells , K562 Cells , Leukemia, Myeloid/pathology , Male , Middle Aged , Polymerase Chain Reaction , RNA, Messenger/analysis , U937 Cells
19.
J Immunol Methods ; 232(1-2): 121-9, 1999 Dec 17.
Article in English | MEDLINE | ID: mdl-10618514

ABSTRACT

In recent years, several investigators have addressed the question of whether mature polymorphonuclear neutrophils (PMN) are able to secrete cytokines. Their studies have brought forward new and exciting discoveries, by establishing that the release of inflammatory cytokines constitutes a novel and important aspect of the neutrophil biology, thereby emphasizing that PMN should no longer be regarded as cells that only release preformed mediators. Although it is still premature to assess the true biological significance of cytokine production by neutrophils, this new aspect of neutrophil biology opens novel perspectives as to the potential role of these cells in the inflammatory and immune responses. In this context, a correct methodological analysis and a detailed molecular investigation of the mechanisms regulating cytokine production by neutrophils in vitro is a critical and fundamental step to better understand how the release of cytokines by PMN may influence pathophysiological processes in vivo. We now describe and discuss the approach that we typically used throughout most of the last decade to characterize cytokine production by human neutrophils, as illustrated herein for a protein that is expressed and released by PMN, namely, vascular endothelial growth factor (VEGF).


Subject(s)
Endothelial Growth Factors/analysis , Lymphokines/analysis , Neutrophils/chemistry , Cells, Cultured , Endothelial Growth Factors/biosynthesis , Endothelial Growth Factors/metabolism , Humans , Interleukin-10/pharmacology , Lipopolysaccharides/pharmacology , Lymphokines/biosynthesis , Lymphokines/metabolism , Neutrophil Activation/immunology , Neutrophils/immunology , Neutrophils/metabolism , Protein Isoforms/analysis , Protein Isoforms/metabolism , RNA, Messenger/biosynthesis , Tumor Necrosis Factor-alpha/pharmacology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors
20.
Blood ; 92(12): 4828-35, 1998 Dec 15.
Article in English | MEDLINE | ID: mdl-9845550

ABSTRACT

Interleukin-2 (IL-2) and IL-15 exert similar biological actions, which largely reflect the fact that their receptors share common beta and gamma subunits; in contrast, distinct subunits are required for high-affinity binding of either cytokine to a heterotrimeric receptor complex. Human neutrophils are known to express both the beta and gamma subunits of the IL-2/IL-15 receptor complex, and we now report that they also constitutively express messenger RNA transcripts encoding the IL-15 receptor chain, suggesting that they possess functional, heterotrimeric IL-15 receptors. Accordingly, we show that in neutrophils, IL-15 elicits several functional responses. In particular, neutrophils synthesize and release IL-8 in response to IL-15, but not to IL-2. Moreover, a nuclear factor-kappaB (NF-kappaB) DNA-binding activity was enhanced in nuclear extracts of IL-15-treated neutrophils, which could be supershifted by antibodies to p50 or RelA. Again, no detectable effect of IL-2 was observed on this response. In peripheral blood lymphocytes (PBL), however, both IL-2 and IL-15 were potent inducers of NF-kappaB activation. Conversely, neither IL-15 nor IL-2 elicited the formation of activator protein-1 (AP-1) DNA-binding complexes in neutrophils, even though both cytokines were found to activate these DNA-binding activities in PBL. Collectively, these observations establish neutrophils as a useful cellular model to discriminate between the actions of IL-15 and IL-2. More importantly, this is the first demonstration that IL-15 has the ability to induce NF-kappaB and AP-1 activation, which further emphasizes the potential relevance of this newly discovered cytokine to immune and inflammatory processes.


Subject(s)
Interleukin-15/pharmacology , Interleukin-8/biosynthesis , NF-kappa B/metabolism , Neutrophils/drug effects , Neutrophils/metabolism , Receptors, Interleukin-2/genetics , Gene Expression , Humans , Interleukin-2/pharmacology , Lymphocytes/drug effects , Lymphocytes/metabolism , Protein Binding , RNA, Messenger/analysis , Receptors, Interleukin-15 , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factor AP-1/metabolism
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