Manuela Mengozzi - Academia.edu (original) (raw)
Papers by Manuela Mengozzi
Journal of Immunology, Aug 1, 1991
The development of LPS tolerance has been suggested to be mediated by an inhibition of cytokine s... more The development of LPS tolerance has been suggested to be mediated by an inhibition of cytokine synthesis. Here we have studied serum IL-6 and TNF levels in mice after LPS administration. Repeated administration of LPS (35 micrograms daily for 4 days) to mice induced a refractoriness (tolerance) to subsequent administrations of LPS in terms of induction of circulating IL-6 and TNF. To investigate the mechanism by which LPS down-regulates its own induction of cytokine synthesis and the relationship between IL-6 and TNF production, we attempted to revert the inhibition of IL-6 and TNF production using agents like PMA or IFN-gamma, previously reported to activate macrophage production of cytokines. Pretreatment with PMA (4 micrograms, 10 min before LPS) partially restored IL-6 production in LPS-tolerant mice given 2 micrograms LPS. On the other hand, PMA did not restore TNF induction in LPS-tolerant mice, even when administered with high doses of LPS (up to 200 micrograms). A similar reversal of LPS resistance to IL-6, but not TNF, induction by PMA was observed in genetically LPS-resistant C3H/HeJ mice. IFN-gamma also restored, although to a lesser extent than PMA, IL-6 production. However, unlike PMA, IFN-gamma could also partially restore TNF production in LPS-tolerant mice, although only when LPS was administered at high doses. By contrast with PMA, IFN-gamma was clearly more active in restoring TNF synthesis than that of IL-6. Similar results were obtained in genetically LPS-unresponsive C3H/HeJ mice. These data suggest that different mechanisms are implicated in the inhibition of IL-6 and TNF synthesis in LPS-tolerant mice and that part of this inhibition can be overcome by PMA or IFN-gamma.
PLOS ONE, May 18, 2015
<p>Proteins identified from BioGEE-treated samples.</p
International Journal of Immunopharmacology, 1991
Interleukin (IL-I) and tumor necrosis factor (TNF) are thought to play a key role in septic shock... more Interleukin (IL-I) and tumor necrosis factor (TNF) are thought to play a key role in septic shock and inflammation. We had previously shown that chlorpromazine (CPZ) has a protective effect in various models of endotoxic shock and IL-I toxicity. We have tested the effect of CPZ on several activities of IL-I in vivo. CPZ (4 mg/kg) inhibited increases in serum corticosterone, triglycerides and serum amyloid A (SAA). Chlorpromazine also antagonized these same effects when they were induced by endotoxin or TNF, suggesting that this activity could be implicated in the protective effect of CPZ in various models of endotoxic shock and IL-1 lethality.
Endocrinology, Mar 1, 1991
Abstract The effect on food intake, body weight, and survival of mice given recombinant lipopolys... more Abstract The effect on food intake, body weight, and survival of mice given recombinant lipopolysaccharide (LPS), tumor necrosis factor/cachectin (TNF), or interleukin 1 (IL-1)(5 μg/mouse, ip, twice daily) was studied. All agents induced a rapid reduction of food intake ...
PubMed, Mar 1, 1993
Cytokine production is down-regulated in LPS tolerance. 1) This down-regulation has been reported... more Cytokine production is down-regulated in LPS tolerance. 1) This down-regulation has been reported in various animal species and cell types, and can be induced both in vivo and in vitro (indicating a desensitization of the cytokine-producing cells). 2) It can also be induced in humans in vivo and in vitro. 3) It is reversible, since the refractory state can be bypassed by administering LPS along with IFN-gamma or PMA. 4) Under certain conditions, it is specific since cytokines will still be induced in response to non-LPS stimuli (Staphylococcus, zymosan, MDP), meaning a real LPS-desensitization is involved rather than an aspecific blockade of cytokine synthesis. 5) in vivo, due to the complexity of the system, other factors than a simple desensitization of cytokine-producing cells contribute to LPS tolerance (aspecific blockade of cytokine synthesis, corticosteroid-dependent feedback, increased clearance by RES and changes in macrophage populations, inhibitory cytokines.
Molecular Medicine, Jun 25, 2020
PLOS ONE, May 18, 2015
<p>Proteins in the NEM-blocked supernatants from BioGEE-pretreated, LPS-stimulated cells we... more <p>Proteins in the NEM-blocked supernatants from BioGEE-pretreated, LPS-stimulated cells were immunoprecipitated with anti-PRDX1 (A) or anti-TXN1 (B). Immunoprecipitated proteins were run under non-reducing (two lanes on the left) or reducing conditions (the two lanes with DTT, on the right). Proteins were then visualized by Western blot with streptavidin peroxidase. The same blot was stripped and reprobed with anti-PRDX1 or anti-TXN1 antibody to locate the proteins (left, in both A and B). m, monomer; d, dimer.</p
PubMed, Oct 1, 1991
A polypeptide homologous to IL-1, with antagonistic activity (IL-1ra), has recently been describe... more A polypeptide homologous to IL-1, with antagonistic activity (IL-1ra), has recently been described. Given the pleiotropic activity of interleukin (IL-1) it is important to establish the spectrum of inhibitory activity of IL-1ra on in vivo action of IL-1. We tested the effect of IL-1ra on IL-1 induced lethality in adrenalectomized mice, hypoglycemia, and induction of serum corticosterone and interleukin 6. Administration of IL-1ra (200 micrograms/mouse) protected adrenalectomized mice against the lethal effect of 1 microgram of IL-1. IL-1ra at 20 micrograms/mouse completely blocked induction of IL-6 and markedly inhibited hypoglycemia and increase in serum corticosterone induced by 0.1 micrograms of IL-1. These data indicate that IL-1ra inhibits the action of IL-1 on different target tissues involved in the various actions of IL-1 in vivo.
Journal of NeuroVirology, 2000
Human immunode®ciency virus (HIV) infection of the central nervous system (CNS) affects primarily... more Human immunode®ciency virus (HIV) infection of the central nervous system (CNS) affects primarily microglial cells and astrocytes. Infection of these latter cells occurs independently of CD4 and is characterised by preferential accumulation of 2 Kb mRNA, encoding mostly Nef, and by low levels of 4.5 and 9 Kb RNAs. We have investigated the potential role of chronic HIV infection of human astrocytic cells on the expression of pro-in¯ammatory cytokines, chemokines and their receptors by comparing the infected TH4-7-5 with its parental uninfected 85HG66 cell lines. Upregulated levels of tumour necrosis factor-a (TNF-a) and of certain chemokines, namely interleukin-8 (IL-8) and regulated upon activation normal T cell expressed and secreted (RANTES), were observed in the infected versus uninfected cells, whereas monocyte chemotactic protein-1 (MCP-1) was comparably expressed in both cell lines. This pattern of expression was con®rmed in primary foetal astrocytes transiently transfected with HIV. In addition, CXCR1, CXCR2 and CCR2b, receptors for IL-8 and MCP-1, respectively, were also found to be upregulated in TH4-7-5 versus 85HG66. CXCR4, the receptor of stromal cell derived factor-1 (SDF-1) and co-receptor for syncytium inducing HIVs, was comparably expressed in infected and uninfected astrocytic cells, whereas CCR5 was not detected in either cell line. Furthermore, treatment of TH4-7-5 cells with TNF-a or IL-1b stimulated RNA and protein secretion of IL-8, MCP-1, and RANTES as well as HIV expression. Thus, our ®ndings suggest that HIV infection of astrocytic cells can contribute to the establishment of a chronic in¯ammatory state in the CNS, eventually resulting in HIV encephalitis, by increasing the secretion of pro-in¯ammatory cytokines, such as TNF-a and several chemokines. Overexpression of chemokine receptors including CCR2b, CXCR1 and CXCR2 in infected astrocytic cells may contribute to HIV-induced damage of the CNS via autocrine/paracrine activation of astrocytes.
Molecular Medicine, Apr 9, 2020
Background: Vitamin D deficiency increases the risk of developing multiple sclerosis (MS) but it ... more Background: Vitamin D deficiency increases the risk of developing multiple sclerosis (MS) but it is unclear whether vitamin D supplementation improves the clinical course of MS, and there is uncertainty about the dose and form of vitamin D (D2 or D3) to be used. The mechanisms underlying the effects of vitamin D in MS are not clear. Vitamin D3 increases the rate of differentiation of primary oligodendrocyte precursor cells (OPCs), suggesting that it might help remyelination in addition to modulating the immune response. Here we analyzed the transcriptome of differentiating rat CG4 OPCs treated with vitamin D2 or with vitamin D3 at 24 h and 72 h following onset of differentiation. Methods: Gene expression in differentiating CG4 cells in response to vitamin D2 or D3 was quantified using Agilent DNA microarrays (n = 4 replicates), and the transcriptome data were processed and analysed using the R software environment. Differential expression between the experimental conditions was determined using LIMMA, applying the Benjamini and Hochberg multiple testing correction to p-values, and significant genes were grouped into coexpression clusters by hierarchical clustering. The functional significance of gene groups was explored by pathway enrichment analysis using the clusterProfiler package. Results: Differentiation alone changed the expression of about 10% of the genes at 72 h compared to 24 h. Vitamin D2 and D3 exerted different effects on gene expression, with D3 influencing 1272 genes and D2 574 at 24 h. The expression of the vast majority of these genes was either not changed in differentiating cells not exposed to vitamin D or followed the same trajectory as the latter. D3-repressed genes were enriched for Gene Ontology (GO) categories including transcription factors and the Notch pathway, while D3-induced genes were enriched for the Ras pathway. Conclusions: This study shows that vitamin D3, compared with D2, changes the expression of a larger number of genes in OLs. Identification of genes affected by D3 in OLs should help to identify mechanisms mediating its action in MS.
Journal of Leukocyte Biology, Oct 1, 2002
Interleukin (IL)-12, especially in the presence of neutralizing anti-IL-4 monoclonal antibodies, ... more Interleukin (IL)-12, especially in the presence of neutralizing anti-IL-4 monoclonal antibodies, primed CD45RO ؊ T clones for high CCL3/macrophage-inflammatory protein-1␣ (MIP-1␣) and CCL4/MIP-1 levels. In CD4 ؉ and CD8 ؉ clones from two patients deficient for IL-12R1 (IL-12R1 ؊/؊), production of CCL3/MIP-1␣ and CCL4/MIP-1 was defective. CD4 ؉ clones from two patients deficient for interferon-␥ (IFN-␥) R1 (IFN-␥R1 ؊/؊) produced somewhat decreased CCL4/MIP-1 levels. IL-12 failed to prime CD4 ؉ or CD8 ؉ healthy clones for high CCL5/regulated on activation, normal T expressed and secreted (RANTES) production, although its secretion was impaired in CD4 ؉ clones from IL-12R1 ؊/؊ and IFN-␥R1 ؊/؊ patients. CCR5 surface expression was up-regulated in resting peripheral blood mononuclear cells and CD4 ؉ clones from both kinds of patients, rendering them more susceptible to CCR5-dependent (R5) HIV-1 infection. Neutralization of IFN-␥ increased CCR5 expression and decreased CC-chemokine secretion by CD4 ؉ clones from healthy and IL-12R1 ؊/؊ individuals, suggesting an IFN-␥-dependent control of CCR5 expression. These data provide the first documented analysis of chemokine secretion and chemokine receptor expression on T cells from IL-12 and IFN-␥ receptor-deficient patients and dissect the role of IL-12 and IFN-␥ on inducing inflammatory chemokine secretion and down-regulating CCR5 expression in human T cells.
Frontiers in Immunology, Feb 5, 2018
The systemic inflammatory response syndrome (SIRS) is a potentially lethal response triggered by ... more The systemic inflammatory response syndrome (SIRS) is a potentially lethal response triggered by diverse forms of tissue injury and infection. When systemic inflammation is triggered by infection, the term sepsis is used. Understanding how inflammation is mediated and regulated is of enormous medical importance. We previously demonstrated that circulating inflammatory-relevant microRNAs (CIR-miRNAs) are candidate biomarkers for differentiating sepsis from SIRS. Here, we set out to determine how CIR-miRNA levels reflect SIRS severity and whether they derive from activated immune cells. Clinical disease severity scores and markers of red blood cell (RBC) damage or immune cell activation were correlated with CIR-miRNA levels in patients with SIRS and sepsis. The release of CIR-miRNAs modulated during SIRS was assessed in immune cell cultures. We show that severity of non-infective SIRS, but not sepsis is reflected in the levels of miR-378a-3p, miR-30a-5p, miR-30d-5p, and miR-192-5p. These CIR-miRNA levels positively correlate with levels of the redox biomarker, peroxiredoxin-1 (Prdx-1), which has previously been shown to be released by immune cells during inflammation. Furthermore, in vitro activated immune cells produce SIRS-associated miR-378a-3p, miR-30a-5p, miR-30d-5p, and miR-192-5p. Our study furthers the understanding of the origin, role, and trafficking of CIR-miRNAs as potential regulators of inflammation.
Cytokine, Nov 1, 2015
Leukaemia inhibitory factor (LIF) and erythropoietin (EPO) are functionally related, neuroprotect... more Leukaemia inhibitory factor (LIF) and erythropoietin (EPO) are functionally related, neuroprotective cytokines. Previously we found that, in rat CG4 oligodendrocyte precursor cells modified to overexpress the EPOR, EPO induces myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) expression in a dose-dependent fashion (0.4–400 ng/ml), as measured by qPCR. In contrast, LIF demonstrates a bell-shaped induction of MOG; a low dose (0.2 ng/ml) increases MOG whereas a high dose (10 ng/ml) is inhibitory. In this study, we sought to define the key intrinsic determinants of LIF-induced myelin gene expression. Inhibition of LIF-induced phospho-STAT3 with stattic downregulated MOG expression. Inhibition of LIF-induced phospho-ERK1/2 with PD184352 as confirmed by western blot, led to a 2-fold increase in MOG at low and high doses of LIF. This was associated with inhibition of LIF-induced early growth response gene 2 (Egr2), a transcription factor which inhibits myelination in this model. In the absence of LIF, differentiation induced basal levels of phospho-ERK1/2 and consequent inhibition increased MBP expression by over 4-fold. High dose LIF also induced 8-fold more suppressor of cytokine signalling 3 (SOCS3) which inversely correlates with the dose-dependent reduction of MOG. In fitting with its linear dose–response, EPO induced comparatively little phospho-STAT3 and SOCS3. These data suggest that the activation of ERK1/2 and the induction of SOCS3 inhibit myelination downstream of the LIF receptor. Overall, the balance of positive and negative intracellular signals determines the myelinogenic capacity of CG4 oligodendrocytes. Pharmacological inhibition of negative signalling pathways could potentiate remyelination in a host of demyelinating diseases.
Biomarker Insights, 2022
Objective: There is an association between frailty and arterial stiffness. However, arterial stif... more Objective: There is an association between frailty and arterial stiffness. However, arterial stiffness does not uniformly correlate with the spectrum of frailty states. Both oxidative stress and inflammaging contribute to vascular ageing. There are no human studies exploring links between arterial stiffness, oxidative stress, inflammaging and frailty. Our objective is to investigate arterial stiffness and inflammaging as predictors of frailty states. Methods: An observational longitudinal cohort study will be used to examine the association between arterial stiffness, oxidative stress and inflammation in 50 older adults (⩾70 years) with clinical frailty scores (CFS) ⩽6 over 6 months. All study measurements will be taken at baseline. Frailty assessment will include hand-grip strength, timed-up and go test, mini-mental state examination, geriatric depression scale and sarcopenia using body composition measurements with Tanita®. Arterial stiffness measurements will include carotid-femoral pulse wave velocity (cfPWV) and carotid-radial pulse wave velocity (crPWV) using Complior (Alam Medical, France). CAVI device will measure Cardio-ankle vascular index and ankle brachial index (ABI). Oxidative stress blood markers nitrotyrosine (NT) and 8-hydroxy-2’-deoxyguanosin (8-oxo-dG) and inflammation markers high-sensitive C-reactive protein (hs-CRP) and interlukin-6(IL-6) will be measured at baseline and 6 month along with lipid profile and glycated haemoglobin. Results (data analysis plan): Descriptive statistics for continuous data using means and standard deviations for normality distributed variables or medians and inter-quartile ranges for skewed variables will be used. Participants will be categorised into CFS 1-3, and CFS 4-6. Categorical data will use frequencies and comparison between groups. Change in frailty between the groups over 6 months will be compared using paired t-test. Simple linear regression will be done between frailty measures, arterial stiffness, inflammation and oxidative stress biomarkers. Significance will be at P < .05. Conclusion: This study data will inform a larger, multi-centre study exploring further the interplay between frailty, biomarkers and arterial stiffness parameters.
PubMed, 1991
Human recombinant IL-1 beta was able to kill C3H/HeJ mice only when inoculated intravenously at v... more Human recombinant IL-1 beta was able to kill C3H/HeJ mice only when inoculated intravenously at very high doses. IL-1 beta, inoculated at 100 mg/kg i.v. as a bolus, induced a shock-like state characterized by anorexia, severe hypothermia and hypoglycemia and persistent neutrophilia, leading to death in 55% of animals generally between 24 and 48 h. In contrast, the noninflammatory adjuvant IL-1 beta peptide VQGEESNDK (position 163-171) did not induce any toxic effect in vivo, when administered following the same schedule. At variance with what was previously observed in endotoxin induced shock, IL-1 beta induced death was not preceded by appearance of circulating TNF. On the other hand, very high and persistent levels of circulating IL-6 could be detected after lethal IL-1 beta administration. Treatment of mice with ibuprofen or with chlorpromazine, both known to counteract some of the toxic effects of IL-1 in vivo, could protect from IL-1 beta induced mortality. Both drugs, at doses protecting from IL-1 beta induced death, were able to abolish IL-1 beta-induced rise of circulating phospholipase A2 (PLA2) activity, and the subsequent generation of toxic PLA2-derived metabolites.
Molecular Immunology, Feb 1, 2002
We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein ... more We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein cysteinyl residues (glutathionylation) in T cell blasts. Treating the cells with the oxidant diamide induces thiolation of a series of proteins that can be detected by 2D electrophoresis when 35 S-cysteine is used to label the intracellular GSH pool. This thiolation is reversible, proteins are rapidly dethiolated and GSH is released from proteins once the oxidants are washed and the cells are allowed to recover. Dethiolation is dependent on the availability of GSH and thiols, since it is inhibited by GSH-depleting agents and improved by N-acetyl-l-cysteine (NAC). The capacity of these agents to reverse glutathionylation is diminished in T cell blasts infected in vitro with HIV, which is known to cause oxidative stress. Consistent with these findings, the activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an enzyme known to be inhibited by glutathionylation, is inhibited in diamide-treated cells and recovers rapidly when cells are allowed to dethiolate. Further, GAPDH activity is diminished by GSH-depleting agents and augmented by NAC. Thus, reversible glutathionylation of proteins can rapidly shift the activity of a key metabolic enzyme and thereby result in dramatic, reversible changes in cellular metabolism.
The FASEB Journal
Several redox modifications have been described during viral infection, including influenza virus... more Several redox modifications have been described during viral infection, including influenza virus infection, but little is known about glutathionylation and this respiratory virus. Glutathionylation is a reversible, post‐translational modification, in which protein cysteine forms transient disulfides with glutathione (GSH), catalyzed by cellular oxidoreductases and in particular by glutaredoxin (Grx). We show here that (i) influenza virus infection induces protein glutathionylation, including that of viral proteins such as hemagglutinin (HA); (ii) Grx1‐mediated deglutathionylation is important for the viral life cycle, as its inhibition, either with an inhibitor of its enzymatic activity or by siRNA, decreases viral replication. Overall these data contribute to the characterization of the complex picture of redox regulation of the influenza virus replication cycle and could help to identify new targets to control respiratory viral infection.
Genes increased by EPO in differentiating cells at 1 h. Genes changed more than 1.5-fold (absolut... more Genes increased by EPO in differentiating cells at 1 h. Genes changed more than 1.5-fold (absolute log2 FC > 0.58), BH adj. P value
Supplementary Figure S1. Effect of EPO on wound closure in BAECs is optimum at a concentration of... more Supplementary Figure S1. Effect of EPO on wound closure in BAECs is optimum at a concentration of 1 ng/ml and after 24 h incubation. Scratch assay model conditions were optimised using different serum concentration (0,1 and 10%) (A) and different EPO concentration (0-100 ng/mL) (B) under 21% or 5% oxygen. Each data point represent mean ± SEM (n=4), *p<0.05, **p<0.01.
Journal of Immunology, Aug 1, 1991
The development of LPS tolerance has been suggested to be mediated by an inhibition of cytokine s... more The development of LPS tolerance has been suggested to be mediated by an inhibition of cytokine synthesis. Here we have studied serum IL-6 and TNF levels in mice after LPS administration. Repeated administration of LPS (35 micrograms daily for 4 days) to mice induced a refractoriness (tolerance) to subsequent administrations of LPS in terms of induction of circulating IL-6 and TNF. To investigate the mechanism by which LPS down-regulates its own induction of cytokine synthesis and the relationship between IL-6 and TNF production, we attempted to revert the inhibition of IL-6 and TNF production using agents like PMA or IFN-gamma, previously reported to activate macrophage production of cytokines. Pretreatment with PMA (4 micrograms, 10 min before LPS) partially restored IL-6 production in LPS-tolerant mice given 2 micrograms LPS. On the other hand, PMA did not restore TNF induction in LPS-tolerant mice, even when administered with high doses of LPS (up to 200 micrograms). A similar reversal of LPS resistance to IL-6, but not TNF, induction by PMA was observed in genetically LPS-resistant C3H/HeJ mice. IFN-gamma also restored, although to a lesser extent than PMA, IL-6 production. However, unlike PMA, IFN-gamma could also partially restore TNF production in LPS-tolerant mice, although only when LPS was administered at high doses. By contrast with PMA, IFN-gamma was clearly more active in restoring TNF synthesis than that of IL-6. Similar results were obtained in genetically LPS-unresponsive C3H/HeJ mice. These data suggest that different mechanisms are implicated in the inhibition of IL-6 and TNF synthesis in LPS-tolerant mice and that part of this inhibition can be overcome by PMA or IFN-gamma.
PLOS ONE, May 18, 2015
<p>Proteins identified from BioGEE-treated samples.</p
International Journal of Immunopharmacology, 1991
Interleukin (IL-I) and tumor necrosis factor (TNF) are thought to play a key role in septic shock... more Interleukin (IL-I) and tumor necrosis factor (TNF) are thought to play a key role in septic shock and inflammation. We had previously shown that chlorpromazine (CPZ) has a protective effect in various models of endotoxic shock and IL-I toxicity. We have tested the effect of CPZ on several activities of IL-I in vivo. CPZ (4 mg/kg) inhibited increases in serum corticosterone, triglycerides and serum amyloid A (SAA). Chlorpromazine also antagonized these same effects when they were induced by endotoxin or TNF, suggesting that this activity could be implicated in the protective effect of CPZ in various models of endotoxic shock and IL-1 lethality.
Endocrinology, Mar 1, 1991
Abstract The effect on food intake, body weight, and survival of mice given recombinant lipopolys... more Abstract The effect on food intake, body weight, and survival of mice given recombinant lipopolysaccharide (LPS), tumor necrosis factor/cachectin (TNF), or interleukin 1 (IL-1)(5 μg/mouse, ip, twice daily) was studied. All agents induced a rapid reduction of food intake ...
PubMed, Mar 1, 1993
Cytokine production is down-regulated in LPS tolerance. 1) This down-regulation has been reported... more Cytokine production is down-regulated in LPS tolerance. 1) This down-regulation has been reported in various animal species and cell types, and can be induced both in vivo and in vitro (indicating a desensitization of the cytokine-producing cells). 2) It can also be induced in humans in vivo and in vitro. 3) It is reversible, since the refractory state can be bypassed by administering LPS along with IFN-gamma or PMA. 4) Under certain conditions, it is specific since cytokines will still be induced in response to non-LPS stimuli (Staphylococcus, zymosan, MDP), meaning a real LPS-desensitization is involved rather than an aspecific blockade of cytokine synthesis. 5) in vivo, due to the complexity of the system, other factors than a simple desensitization of cytokine-producing cells contribute to LPS tolerance (aspecific blockade of cytokine synthesis, corticosteroid-dependent feedback, increased clearance by RES and changes in macrophage populations, inhibitory cytokines.
Molecular Medicine, Jun 25, 2020
PLOS ONE, May 18, 2015
<p>Proteins in the NEM-blocked supernatants from BioGEE-pretreated, LPS-stimulated cells we... more <p>Proteins in the NEM-blocked supernatants from BioGEE-pretreated, LPS-stimulated cells were immunoprecipitated with anti-PRDX1 (A) or anti-TXN1 (B). Immunoprecipitated proteins were run under non-reducing (two lanes on the left) or reducing conditions (the two lanes with DTT, on the right). Proteins were then visualized by Western blot with streptavidin peroxidase. The same blot was stripped and reprobed with anti-PRDX1 or anti-TXN1 antibody to locate the proteins (left, in both A and B). m, monomer; d, dimer.</p
PubMed, Oct 1, 1991
A polypeptide homologous to IL-1, with antagonistic activity (IL-1ra), has recently been describe... more A polypeptide homologous to IL-1, with antagonistic activity (IL-1ra), has recently been described. Given the pleiotropic activity of interleukin (IL-1) it is important to establish the spectrum of inhibitory activity of IL-1ra on in vivo action of IL-1. We tested the effect of IL-1ra on IL-1 induced lethality in adrenalectomized mice, hypoglycemia, and induction of serum corticosterone and interleukin 6. Administration of IL-1ra (200 micrograms/mouse) protected adrenalectomized mice against the lethal effect of 1 microgram of IL-1. IL-1ra at 20 micrograms/mouse completely blocked induction of IL-6 and markedly inhibited hypoglycemia and increase in serum corticosterone induced by 0.1 micrograms of IL-1. These data indicate that IL-1ra inhibits the action of IL-1 on different target tissues involved in the various actions of IL-1 in vivo.
Journal of NeuroVirology, 2000
Human immunode®ciency virus (HIV) infection of the central nervous system (CNS) affects primarily... more Human immunode®ciency virus (HIV) infection of the central nervous system (CNS) affects primarily microglial cells and astrocytes. Infection of these latter cells occurs independently of CD4 and is characterised by preferential accumulation of 2 Kb mRNA, encoding mostly Nef, and by low levels of 4.5 and 9 Kb RNAs. We have investigated the potential role of chronic HIV infection of human astrocytic cells on the expression of pro-in¯ammatory cytokines, chemokines and their receptors by comparing the infected TH4-7-5 with its parental uninfected 85HG66 cell lines. Upregulated levels of tumour necrosis factor-a (TNF-a) and of certain chemokines, namely interleukin-8 (IL-8) and regulated upon activation normal T cell expressed and secreted (RANTES), were observed in the infected versus uninfected cells, whereas monocyte chemotactic protein-1 (MCP-1) was comparably expressed in both cell lines. This pattern of expression was con®rmed in primary foetal astrocytes transiently transfected with HIV. In addition, CXCR1, CXCR2 and CCR2b, receptors for IL-8 and MCP-1, respectively, were also found to be upregulated in TH4-7-5 versus 85HG66. CXCR4, the receptor of stromal cell derived factor-1 (SDF-1) and co-receptor for syncytium inducing HIVs, was comparably expressed in infected and uninfected astrocytic cells, whereas CCR5 was not detected in either cell line. Furthermore, treatment of TH4-7-5 cells with TNF-a or IL-1b stimulated RNA and protein secretion of IL-8, MCP-1, and RANTES as well as HIV expression. Thus, our ®ndings suggest that HIV infection of astrocytic cells can contribute to the establishment of a chronic in¯ammatory state in the CNS, eventually resulting in HIV encephalitis, by increasing the secretion of pro-in¯ammatory cytokines, such as TNF-a and several chemokines. Overexpression of chemokine receptors including CCR2b, CXCR1 and CXCR2 in infected astrocytic cells may contribute to HIV-induced damage of the CNS via autocrine/paracrine activation of astrocytes.
Molecular Medicine, Apr 9, 2020
Background: Vitamin D deficiency increases the risk of developing multiple sclerosis (MS) but it ... more Background: Vitamin D deficiency increases the risk of developing multiple sclerosis (MS) but it is unclear whether vitamin D supplementation improves the clinical course of MS, and there is uncertainty about the dose and form of vitamin D (D2 or D3) to be used. The mechanisms underlying the effects of vitamin D in MS are not clear. Vitamin D3 increases the rate of differentiation of primary oligodendrocyte precursor cells (OPCs), suggesting that it might help remyelination in addition to modulating the immune response. Here we analyzed the transcriptome of differentiating rat CG4 OPCs treated with vitamin D2 or with vitamin D3 at 24 h and 72 h following onset of differentiation. Methods: Gene expression in differentiating CG4 cells in response to vitamin D2 or D3 was quantified using Agilent DNA microarrays (n = 4 replicates), and the transcriptome data were processed and analysed using the R software environment. Differential expression between the experimental conditions was determined using LIMMA, applying the Benjamini and Hochberg multiple testing correction to p-values, and significant genes were grouped into coexpression clusters by hierarchical clustering. The functional significance of gene groups was explored by pathway enrichment analysis using the clusterProfiler package. Results: Differentiation alone changed the expression of about 10% of the genes at 72 h compared to 24 h. Vitamin D2 and D3 exerted different effects on gene expression, with D3 influencing 1272 genes and D2 574 at 24 h. The expression of the vast majority of these genes was either not changed in differentiating cells not exposed to vitamin D or followed the same trajectory as the latter. D3-repressed genes were enriched for Gene Ontology (GO) categories including transcription factors and the Notch pathway, while D3-induced genes were enriched for the Ras pathway. Conclusions: This study shows that vitamin D3, compared with D2, changes the expression of a larger number of genes in OLs. Identification of genes affected by D3 in OLs should help to identify mechanisms mediating its action in MS.
Journal of Leukocyte Biology, Oct 1, 2002
Interleukin (IL)-12, especially in the presence of neutralizing anti-IL-4 monoclonal antibodies, ... more Interleukin (IL)-12, especially in the presence of neutralizing anti-IL-4 monoclonal antibodies, primed CD45RO ؊ T clones for high CCL3/macrophage-inflammatory protein-1␣ (MIP-1␣) and CCL4/MIP-1 levels. In CD4 ؉ and CD8 ؉ clones from two patients deficient for IL-12R1 (IL-12R1 ؊/؊), production of CCL3/MIP-1␣ and CCL4/MIP-1 was defective. CD4 ؉ clones from two patients deficient for interferon-␥ (IFN-␥) R1 (IFN-␥R1 ؊/؊) produced somewhat decreased CCL4/MIP-1 levels. IL-12 failed to prime CD4 ؉ or CD8 ؉ healthy clones for high CCL5/regulated on activation, normal T expressed and secreted (RANTES) production, although its secretion was impaired in CD4 ؉ clones from IL-12R1 ؊/؊ and IFN-␥R1 ؊/؊ patients. CCR5 surface expression was up-regulated in resting peripheral blood mononuclear cells and CD4 ؉ clones from both kinds of patients, rendering them more susceptible to CCR5-dependent (R5) HIV-1 infection. Neutralization of IFN-␥ increased CCR5 expression and decreased CC-chemokine secretion by CD4 ؉ clones from healthy and IL-12R1 ؊/؊ individuals, suggesting an IFN-␥-dependent control of CCR5 expression. These data provide the first documented analysis of chemokine secretion and chemokine receptor expression on T cells from IL-12 and IFN-␥ receptor-deficient patients and dissect the role of IL-12 and IFN-␥ on inducing inflammatory chemokine secretion and down-regulating CCR5 expression in human T cells.
Frontiers in Immunology, Feb 5, 2018
The systemic inflammatory response syndrome (SIRS) is a potentially lethal response triggered by ... more The systemic inflammatory response syndrome (SIRS) is a potentially lethal response triggered by diverse forms of tissue injury and infection. When systemic inflammation is triggered by infection, the term sepsis is used. Understanding how inflammation is mediated and regulated is of enormous medical importance. We previously demonstrated that circulating inflammatory-relevant microRNAs (CIR-miRNAs) are candidate biomarkers for differentiating sepsis from SIRS. Here, we set out to determine how CIR-miRNA levels reflect SIRS severity and whether they derive from activated immune cells. Clinical disease severity scores and markers of red blood cell (RBC) damage or immune cell activation were correlated with CIR-miRNA levels in patients with SIRS and sepsis. The release of CIR-miRNAs modulated during SIRS was assessed in immune cell cultures. We show that severity of non-infective SIRS, but not sepsis is reflected in the levels of miR-378a-3p, miR-30a-5p, miR-30d-5p, and miR-192-5p. These CIR-miRNA levels positively correlate with levels of the redox biomarker, peroxiredoxin-1 (Prdx-1), which has previously been shown to be released by immune cells during inflammation. Furthermore, in vitro activated immune cells produce SIRS-associated miR-378a-3p, miR-30a-5p, miR-30d-5p, and miR-192-5p. Our study furthers the understanding of the origin, role, and trafficking of CIR-miRNAs as potential regulators of inflammation.
Cytokine, Nov 1, 2015
Leukaemia inhibitory factor (LIF) and erythropoietin (EPO) are functionally related, neuroprotect... more Leukaemia inhibitory factor (LIF) and erythropoietin (EPO) are functionally related, neuroprotective cytokines. Previously we found that, in rat CG4 oligodendrocyte precursor cells modified to overexpress the EPOR, EPO induces myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) expression in a dose-dependent fashion (0.4–400 ng/ml), as measured by qPCR. In contrast, LIF demonstrates a bell-shaped induction of MOG; a low dose (0.2 ng/ml) increases MOG whereas a high dose (10 ng/ml) is inhibitory. In this study, we sought to define the key intrinsic determinants of LIF-induced myelin gene expression. Inhibition of LIF-induced phospho-STAT3 with stattic downregulated MOG expression. Inhibition of LIF-induced phospho-ERK1/2 with PD184352 as confirmed by western blot, led to a 2-fold increase in MOG at low and high doses of LIF. This was associated with inhibition of LIF-induced early growth response gene 2 (Egr2), a transcription factor which inhibits myelination in this model. In the absence of LIF, differentiation induced basal levels of phospho-ERK1/2 and consequent inhibition increased MBP expression by over 4-fold. High dose LIF also induced 8-fold more suppressor of cytokine signalling 3 (SOCS3) which inversely correlates with the dose-dependent reduction of MOG. In fitting with its linear dose–response, EPO induced comparatively little phospho-STAT3 and SOCS3. These data suggest that the activation of ERK1/2 and the induction of SOCS3 inhibit myelination downstream of the LIF receptor. Overall, the balance of positive and negative intracellular signals determines the myelinogenic capacity of CG4 oligodendrocytes. Pharmacological inhibition of negative signalling pathways could potentiate remyelination in a host of demyelinating diseases.
Biomarker Insights, 2022
Objective: There is an association between frailty and arterial stiffness. However, arterial stif... more Objective: There is an association between frailty and arterial stiffness. However, arterial stiffness does not uniformly correlate with the spectrum of frailty states. Both oxidative stress and inflammaging contribute to vascular ageing. There are no human studies exploring links between arterial stiffness, oxidative stress, inflammaging and frailty. Our objective is to investigate arterial stiffness and inflammaging as predictors of frailty states. Methods: An observational longitudinal cohort study will be used to examine the association between arterial stiffness, oxidative stress and inflammation in 50 older adults (⩾70 years) with clinical frailty scores (CFS) ⩽6 over 6 months. All study measurements will be taken at baseline. Frailty assessment will include hand-grip strength, timed-up and go test, mini-mental state examination, geriatric depression scale and sarcopenia using body composition measurements with Tanita®. Arterial stiffness measurements will include carotid-femoral pulse wave velocity (cfPWV) and carotid-radial pulse wave velocity (crPWV) using Complior (Alam Medical, France). CAVI device will measure Cardio-ankle vascular index and ankle brachial index (ABI). Oxidative stress blood markers nitrotyrosine (NT) and 8-hydroxy-2’-deoxyguanosin (8-oxo-dG) and inflammation markers high-sensitive C-reactive protein (hs-CRP) and interlukin-6(IL-6) will be measured at baseline and 6 month along with lipid profile and glycated haemoglobin. Results (data analysis plan): Descriptive statistics for continuous data using means and standard deviations for normality distributed variables or medians and inter-quartile ranges for skewed variables will be used. Participants will be categorised into CFS 1-3, and CFS 4-6. Categorical data will use frequencies and comparison between groups. Change in frailty between the groups over 6 months will be compared using paired t-test. Simple linear regression will be done between frailty measures, arterial stiffness, inflammation and oxidative stress biomarkers. Significance will be at P < .05. Conclusion: This study data will inform a larger, multi-centre study exploring further the interplay between frailty, biomarkers and arterial stiffness parameters.
PubMed, 1991
Human recombinant IL-1 beta was able to kill C3H/HeJ mice only when inoculated intravenously at v... more Human recombinant IL-1 beta was able to kill C3H/HeJ mice only when inoculated intravenously at very high doses. IL-1 beta, inoculated at 100 mg/kg i.v. as a bolus, induced a shock-like state characterized by anorexia, severe hypothermia and hypoglycemia and persistent neutrophilia, leading to death in 55% of animals generally between 24 and 48 h. In contrast, the noninflammatory adjuvant IL-1 beta peptide VQGEESNDK (position 163-171) did not induce any toxic effect in vivo, when administered following the same schedule. At variance with what was previously observed in endotoxin induced shock, IL-1 beta induced death was not preceded by appearance of circulating TNF. On the other hand, very high and persistent levels of circulating IL-6 could be detected after lethal IL-1 beta administration. Treatment of mice with ibuprofen or with chlorpromazine, both known to counteract some of the toxic effects of IL-1 in vivo, could protect from IL-1 beta induced mortality. Both drugs, at doses protecting from IL-1 beta induced death, were able to abolish IL-1 beta-induced rise of circulating phospholipase A2 (PLA2) activity, and the subsequent generation of toxic PLA2-derived metabolites.
Molecular Immunology, Feb 1, 2002
We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein ... more We show here that exposure to oxidative stress induces glutathione (GSH) modification of protein cysteinyl residues (glutathionylation) in T cell blasts. Treating the cells with the oxidant diamide induces thiolation of a series of proteins that can be detected by 2D electrophoresis when 35 S-cysteine is used to label the intracellular GSH pool. This thiolation is reversible, proteins are rapidly dethiolated and GSH is released from proteins once the oxidants are washed and the cells are allowed to recover. Dethiolation is dependent on the availability of GSH and thiols, since it is inhibited by GSH-depleting agents and improved by N-acetyl-l-cysteine (NAC). The capacity of these agents to reverse glutathionylation is diminished in T cell blasts infected in vitro with HIV, which is known to cause oxidative stress. Consistent with these findings, the activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an enzyme known to be inhibited by glutathionylation, is inhibited in diamide-treated cells and recovers rapidly when cells are allowed to dethiolate. Further, GAPDH activity is diminished by GSH-depleting agents and augmented by NAC. Thus, reversible glutathionylation of proteins can rapidly shift the activity of a key metabolic enzyme and thereby result in dramatic, reversible changes in cellular metabolism.
The FASEB Journal
Several redox modifications have been described during viral infection, including influenza virus... more Several redox modifications have been described during viral infection, including influenza virus infection, but little is known about glutathionylation and this respiratory virus. Glutathionylation is a reversible, post‐translational modification, in which protein cysteine forms transient disulfides with glutathione (GSH), catalyzed by cellular oxidoreductases and in particular by glutaredoxin (Grx). We show here that (i) influenza virus infection induces protein glutathionylation, including that of viral proteins such as hemagglutinin (HA); (ii) Grx1‐mediated deglutathionylation is important for the viral life cycle, as its inhibition, either with an inhibitor of its enzymatic activity or by siRNA, decreases viral replication. Overall these data contribute to the characterization of the complex picture of redox regulation of the influenza virus replication cycle and could help to identify new targets to control respiratory viral infection.
Genes increased by EPO in differentiating cells at 1 h. Genes changed more than 1.5-fold (absolut... more Genes increased by EPO in differentiating cells at 1 h. Genes changed more than 1.5-fold (absolute log2 FC > 0.58), BH adj. P value
Supplementary Figure S1. Effect of EPO on wound closure in BAECs is optimum at a concentration of... more Supplementary Figure S1. Effect of EPO on wound closure in BAECs is optimum at a concentration of 1 ng/ml and after 24 h incubation. Scratch assay model conditions were optimised using different serum concentration (0,1 and 10%) (A) and different EPO concentration (0-100 ng/mL) (B) under 21% or 5% oxygen. Each data point represent mean ± SEM (n=4), *p<0.05, **p<0.01.