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Papers by Maria Borghi

Mitochondria morphology in mutant TARDBP and C9ORF72 fibroblasts after 48 h in galactose medium. ... more Mitochondria morphology in mutant TARDBP and C9ORF72 fibroblasts after 48 h in galactose medium. Representative confocal images of mitochondrial network in primary fibroblasts from three healthy controls (C1, C2, C3), three ALS patients carrying mutation in TARDBP gene (p.A382T; T1, T2, T3) and three ALS//FTD patients with pathological expansion of the hexanucleotide repeat in C9ORF72 (C9.2, C9.3, C9.4) transfected with pDsRed2Mito construct. Bar, 10μm. Figure S2. Mitochondria functionality in mutant TARDBP and C9ORF72 fibroblasts. Measurement of mitochondrial membrane potential by flow cytometry analysis of Mitotracker Red (MTR) positive fibroblasts (from 4 healthy controls, 3 TARDBP and 4 C9ORF72 mutated patients) maintained in medium with galactose and without glucose. Median ± SEM, n = 3 different DIV; One-way ANOVA with Dunnett's multiple comparison test; **p

International Journal of Molecular Sciences, 2021

Medullary thyroid carcinoma (MTC) is a tumor deriving from the thyroid C cells. Vandetanib (VAN) ... more Medullary thyroid carcinoma (MTC) is a tumor deriving from the thyroid C cells. Vandetanib (VAN) and cabozantinib (CAB) are two tyrosine kinase inhibitors targeting REarranged during Transfection (RET) and other kinase receptors and are approved for the treatment of advanced MTC. We aim to compare the in vitro and in vivo anti-tumor activity of VAN and CAB in MTC. The effects of VAN and CAB on viability, cell cycle, and apoptosis of TT and MZ-CRC-1 cells are evaluated in vitro using an MTT assay, DNA flow cytometry with propidium iodide, and Annexin V-FITC/propidium iodide staining, respectively. In vivo, the anti-angiogenic potential of VAN and CAB is evaluated in Tg(fli1a:EGFP)y1 transgenic fluorescent zebrafish embryos by analyzing the effects on the physiological development of the sub-intestinal vein plexus and the tumor-induced angiogenesis after TT and MZ-CRC-1 xenotransplantation. VAN and CAB exert comparable effects on TT and MZ-CRC-1 viability inhibition and cell cycle per...

Journal of Translational Autoimmunity, 2021

Arthritis Research & Therapy, 2020

Background Consistently with their diagnostic and prognostic value, autoantibodies specific for s... more Background Consistently with their diagnostic and prognostic value, autoantibodies specific for systemic sclerosis (SSc) embedded in immune complexes (ICs) elicited a pro-inflammatory and pro-fibrotic cascade in healthy skin fibroblasts, engaging Toll-like receptors (TLRs) via their nucleic acid components. The objective of this study was to investigate the pathogenicity of SSc-ICs in endothelial cells. Methods ICs were purified from the sera of SSc patients bearing different autoantibody specificities (antibodies against DNA topoisomerase I, centromeric proteins, RNA polymerase, and Th/To), patients with systemic lupus erythematosus (SLE) and primary anti-phospholipid syndrome (PAPS), or healthy controls (NHS) using polyethylene glycol precipitation. Human umbilical vein endothelial cells (HUVECs) were incubated with ICs, positive and negative controls. mRNA levels of endothelin-1 (et-1), collagenIα1 (colIα1), interferon (IFN)-α, and IFN-β were investigated by real-time PCR; et-1 a...

The Israel Medical Association journal : IMAJ, 2002

Systemic sclerosis is a disease characterized by fibrosis of skin and internal organs. Although a... more Systemic sclerosis is a disease characterized by fibrosis of skin and internal organs. Although at present the classical criteria defining an autoimmune disease are not entirely fulfilled for SSc, several autoimmune abnormalities, both cellular and humoral, have been ...

Arthritis & Rheumatology, 2015

Objective: Rheumatoid arthritis (RA) is associated with accelerated atherosclerosis. The reductio... more Objective: Rheumatoid arthritis (RA) is associated with accelerated atherosclerosis. The reduction of cardiovascular risk induced by methotrexate and anti-TNFα agents in RA is considered secondary to their anti-inflammatory action, but their effects on serum lipoprotein function and foam cell formation are unknown. The reduced high density lipoprotein (HDL) capacity to promote cell cholesterol efflux (CEC) and the increased serum cell cholesterol-loading capacity (CLC) demonstrated in RA may contribute to foam cell development. We investigated the influence of methotrexate and adalimumab treatment on serum CEC and CLC in RA patients. Furthermore, we studied the in vitro effect of the two drugs on macrophage cholesterol handling. Methods: serum from RA patients treated with methotrexate (n=34) or with adalimumab+methotrexate (n=22), drawn before and after 6 weeks and 6 months of treatment, was analyzed for CEC and CLC by radioisotopic and fluorimetric techniques, respectively. Methotrexate and adalimumab influence on macrophage cholesterol efflux and uptake was evaluated in vitro on human THP-1-derived macrophages. Results: methotrexate treatment was associated with serum HDL, LDL and total cholesterol increase, with ATP-binding cassette G1 and Scavenger receptor-BI (SR-BI)-mediated CEC increase and no CLC modification. Adalimumab treatment was associated with serum HDL increase, SR-BI-mediated and ATP-binding cassette A1-mediated CEC transient increase and decrease respectively, and with a significant CLC reduction; in addition, adalimumab reduced macrophage cholesterol uptake in vitro. Conclusion: methotrexate and adalimumab anti-atherosclerotic activity may be mediated by beneficial and complementary effects on lipoprotein functions and on macrophage cholesterol handling, as a whole opposing foam cell formation. This article is protected by copyright. All rights reserved.

Annals of the rheumatic diseases, 2014

The marked cardiovascular risk in autoimmune diseases is only partly explained. The capacity of h... more The marked cardiovascular risk in autoimmune diseases is only partly explained. The capacity of high-density lipoproteins (HDL) to promote cell cholesterol efflux is a property with a well-known anti-atherogenic significance, but is also involved in functional modulation of endothelial and immune cells. The aim of this work was to evaluate HDL functionality with respect to cell cholesterol efflux in rheumatoid arthritis (RA) and systemic lupus erythemathosus (SLE) patients. We evaluated serum cholesterol efflux capacity (CEC) of apoB-depleted serum, which mainly reflects HDL activity, from 30 RA and 30 SLE patients, and from 30 healthy controls by radioisotopic ex-vivo systems discriminating between the specific pathways of cholesterol efflux. RA patients presented impairment of ATP-binding cassette G1-mediated CEC that correlated with disease activity. SLE patients showed a more complex pattern of modifications unrelated to disease activity, with marked reduction of ATP-binding cas...

Trends in Immunology, 2005

Despite the discovery of anti-endothelial cell antibodies (AECA) in a heterogeneous group of diso... more Despite the discovery of anti-endothelial cell antibodies (AECA) in a heterogeneous group of disorders characterized by endothelial damage, their pathogenic role is still debated. Experimental in vitro models indicate that they can either damage endothelial cells or trigger cell signaling by reacting with as yet undefined surface molecules. However, clinical studies suggest that, in addition to AECA, other pathogenic mechanisms are involved in the vasculitic process. Recently, antibodies specific for b2 glycoprotein I, the phospholipid-binding protein targeted by anti-phospholipid antibodies, have been shown to display anti-endothelial activity. These autoantibodies recognize b2 glycoprotein I adhered to the endothelium and induce a cell perturbation that might underlie the thrombophilic state of the antiphospholipid syndrome.

Lupus, 2007

The administration of immunosuppressive drugs during pregnancy is often necessary in women with a... more The administration of immunosuppressive drugs during pregnancy is often necessary in women with autoimmune diseases. Teratogenicity of immunosuppressives during pregnancy has been evaluated, only few data exist about the effects on immune systems. We therefore performed a pilot study on the influence of foetal exposure to immunosuppressives on immune function of babies born to mothers with autoimmune disorders. We investigated serological and cellular parameters as indicators of immune system status. We included in the study 14 babies (mean age 11 months, range 1—24) born to mothers with autoimmune diseases and exposed in utero to different immunosuppressants and, as controls, 14 babies whose mothers had autoimmune manifestations but did not receive immunosuppressive therapy. We evaluated: (i) complete blood count, (ii) immunoglobulin levels and IgG subclasses, (iii) antibody response to hepatitis B vaccine, (iv) leukocyte subpopulations and (v) interleukin-2 and interferon γ in vit...

Journal of Cellular and Molecular Medicine, 2012

Nature reviews. Rheumatology, 2018

Recurrent thrombosis and miscarriages are the main clinical manifestations of antiphospholipid sy... more Recurrent thrombosis and miscarriages are the main clinical manifestations of antiphospholipid syndrome (APS). Although most patients display both clinical signs, some patients can have isolated vascular or obstetric variants. Emerging data raise the question of whether obstetric and vascular APS are the same or different diseases. An important difference between the two conditions is that a thrombophilic state is a common feature in vascular APS, whereas clot occlusions of the decidual spiral arteries are seldom observed in obstetric APS, and infarctions are found in only one-third of APS placentae. Conversely, inflammation, which is undetectable in vascular APS, is frequently observed in the placentae of patients with obstetric APS and has been documented in the placentae of pregnant mice with fetal loss mediated by antiphospholipid antibodies. Attempts to identify different antibodies or epitopes responsible for the two clinical manifestations of APS have so far been unsuccessful...

PloS one, 2017

Domain I (DI) of beta-2-glycoprotein I (β2GPI) contains the immunodominant epitope for pathogenic... more Domain I (DI) of beta-2-glycoprotein I (β2GPI) contains the immunodominant epitope for pathogenic antiphospholipid antibodies (aPL). DI is exposed in the linear form of the molecule but not in the circular form that comprises 90% of serum β2GPI. The majority of circulating β2GPI is biochemically reduced with two free thiols in Domain V. However, increased levels of oxidised β2GPI are found in patients with antiphospholipid syndrome (APS). It is not known whether oxidation of β2GPI favours the linear form of the molecule and thus promotes development of anti-DI antibodies. We investigated whether the proportion of oxidised β2GPI associates with the presence of anti-DI in APS patients. Serum samples from 44 APS patients were screened for IgG, IgM and IgA anti-DI, anti-β2GPI, anti-cardiolipin (anti-CL) and biochemically reduced β2GPI. A negative correlation was found between the proportion of β2GPI in the biochemically reduced form and IgG anti-DI levels (r = -0.54, p = 0.0002), but no...

Microvascular research, 2017

Raynaud's phenomenon (RP) can be the first manifestation of systemic sclerosis (SSc) or other... more Raynaud's phenomenon (RP) can be the first manifestation of systemic sclerosis (SSc) or other connective tissue diseases (CTDs), often preceding an overt disease by years. It is not known if markers of endothelial damage are detectable in those RP patients who subsequently develop a CTD. We studied 82 RP patients at their first evaluation to correlate the levels of endothelial markers with the subsequent development of an overt disease 36months later. We measured plasma levels of tissue-type plasminogen activator (t-PA) and von Willebrand factor (vWF), two markers of endothelial damage, and interleukin-6 (IL-6), a pro-inflammatory cytokine. Thirty sex- and age-matched healthy subjects (HS) served as controls. At baseline, 67 patients showed capillaroscopic normal pattern (CNP) and 15 patients, of which 11 were very early SSc, had capillaroscopic scleroderma pattern (CSP). Plasma levels of t-PA, vWF and IL-6 were higher in patients with CNP (p=0.0001) than in HS and even much hig...

Molecular oncology, Jan 28, 2017

Medullary thyroid cancer (MTC) is a tumor highly resistant to chemo- and radiotherapy. Drug resis... more Medullary thyroid cancer (MTC) is a tumor highly resistant to chemo- and radiotherapy. Drug resistance can be induced by epigenetic changes such as aberrant DNA methylation. To overcome drug resistance, we explored a promising approach based on the use of 5-aza-2'-deoxycytidine (AZA), a demethylating agent, in combination with the mTOR inhibitor everolimus in MTC cells (MZ-CRC-1 and TT). This combined treatment showed a strong synergistic antiproliferative activity through the induction of apoptosis. The effect of everolimus and/or AZA on genome-wide expression profiling was evaluated by Illumina BeadChip in MZ-CRC-1 cells. An innovative bioinformatic pipeline identified four potential molecular pathways implicated in the synergy between AZA and everolimus: PI3K-Akt signaling, the neurotrophin pathway, ECM-receptor interaction and focal adhesion. Among these, the neurotrophin signaling pathway was most directly involved in apoptosis, through the overexpression of NGFR and Bax ge...

The Journal of Immunology, 2017

Journal of acquired immune deficiency syndromes

Sera from patients affected either by the classic/Mediterranean form (5 patients) and human immun... more Sera from patients affected either by the classic/Mediterranean form (5 patients) and human immune deficiency virus (HIV)-associated (12 patients) form of Kaposi's sarcoma (KS) were compared in supporting endothelial cell in-vitro proliferation. Healthy blood donors (15) and a group of 7 HIV-positive drug addicts with no dermatological involvement were used as control groups. Endothelial cell growth was assessed by means of a [3H]thymidine incorporation and by an optical density direct cell-counting assay. Our results indicate that, at the highest concentrations, sera obtained from patients affected by KS with no sign of HIV infection induced significantly higher levels of endothelial cell proliferation when compared with HIV-associated KS and with normal controls. This growth-promoting activity was apparently cell selective, being present in endothelial cell but not in fibroblast cultures.

Journal of clinical & laboratory immunology

The present report investigates the synergistic effects of different agents (phytohaemoagglutinin... more The present report investigates the synergistic effects of different agents (phytohaemoagglutinin (PHA), anti-CD3, phorbol-12-myristate-13-acetate (PMA) and Ionomycin on T cell activation. Results indicate that in normal lymphocytes PMA causes a dose-dependent decrease of PHA-induced blastogenesis and a marked enhancement of anti-CD3-induced 3H-thymidine uptake. There is positive synergism when cells are costimulated with PMA and the calcium ionophore Ionomycin. These data show that phorbol esters can exert either positive or negative synergisms depending on the experimental conditions. We also studied the simultaneous stimulation with PHA and anti-CD3 which gives an addictive effect for the minimal doses of the two stimuli, and an appreciable negative synergism for their highest concentrations. In order to evaluate T lymphocyte functions in the age-associated immune defect, PHA plus anti-CD3 and PMA plus Ionomycin costimulation assays were applied to a population of healthy elderly subjects. An additive effect is found after costimulation with the highest doses of PHA and anti-CD3, at variance with the negative synergism found in younger adult controls. Following costimulation with PMA plus Ionomycin, elderly subjects also display an impaired response, as compared to adult controls. No correlation was found with PHA-, anti-CD3- and PMA plus Ionomycin-induced T cell proliferation. These results underline the complexity of age-associated immune defects and suggest that alterations in different mechanisms of lymphocyte activation are responsible for the immune deficiency of aging.

Infection

The biological response modifying activities of cefodizime (CDZ), a new third-generation cephalos... more The biological response modifying activities of cefodizime (CDZ), a new third-generation cephalosporin, were investigated in vitro and ex vivo. In vitro investigations using cells isolated from the blood of young healthy donors showed no stimulating activity of CDZ on peripheral blood lymphocytes, natural killer cell activity, IL-1 production by adherent mononuclear cells, PMN chemiluminescence or PMN chemotaxis. A slight but statistically insignificant increase in PMN phagocytosis and phagocytic index was observed in the same population. IL-1 production was increased in three subjects with low resting state values. In a controlled ex vivo study, 20 healthy elderly subjects selected on the basis of depressed phagocytic function were treated with CDZ 1 g i.m. b.i.d. or placebo for eight days. PMN function was determined at baseline and on the day after the last dose. In the CDZ group a significant increase in both phagocytosis and phagocytic index was found, while there were no chang...

Annals of the Rheumatic Diseases, 2014

ABSTRACT Background Raynaud's phenomenon (RP) may predate an overt Connective Tissue Dise... more ABSTRACT Background Raynaud's phenomenon (RP) may predate an overt Connective Tissue Disease (CTD) of the “Scleroderma spectrum” such as Systemic sclerosis (SSc). The predictive role of auto-antibodies (auto-Abs) such as ANA for the development of a CTD in patients with isolated RP (i.e. with no other signs or symptoms of CTD) is well known, whereas data regarding the prevalence of anti-ENA in such patients are lacking. Objectives Our aim was to evaluate the profile of anti-ENA in patients with isolated RP. Methods Sera were analyzed for anti-ENA by EUROLINE “Scleroderma profile” (antigens: Scl70, CENP A, CENP B, RP11, RP155, fibrillarin (Fib), NOR90, Th/To, PM-Scl100, PM-Scl75, Ku, PDGFR, Ro-52) and “Myosites profile” (antigens: Mi-2, Ku, PM-Scl100, PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ, Ro-52) [EUROIMMUN AG Leuback, Germany]. Results A total of 115 adults (105 females and 10 males) with isolated RP were enrolled in two Italian Rheumatology Centers (G. Pini Institute, Milan and Moriggia-Pelascini Hospital, Gravedona) from March 2011 to September 2013. Forty-four of 115 (38.0%) subjects were positive for at least one anti-ENA; 71 out of 110 (61.7%) were negative both for Scleroderma and Myosites profiles (see table 1 and 2). Conclusions Up to 38% of RP subjects were positive for at least one anti-ENA. The detection of anti-ENA since the first Rheumatologic evaluation may aid the specialist in achieving an earlier diagnosis. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5955

Arthritis & Rheumatology, 2015

It was suggested that only antibodies against Domain 1 (D1) of β2glycoprotein I (β2 GPI) are path... more It was suggested that only antibodies against Domain 1 (D1) of β2glycoprotein I (β2 GPI) are pathogenic and diagnostic. Still debated is the role of antibodies against other β2GPI domains. Our aim was to evaluate the clinical relevance of domain specificity profiling of anti-β2GPI IgG antibodies not only in Antiphospholipid Syndrome (APS) patients but also in control groups of Systemic Autoimmune Rheumatic Diseases (SARDs) and asymptomatic Antiphospholipid Antibody (aPL) carriers. 159 subjects with persistently positive, medium/high titer anti-β2GPI IgG were included: 1) 56 thrombotic (+/- obstetric) primary APS (PAPS); 2) 31 women with pure obstetric PAPS; 3) 42 aPL-positive SARDs; 4) 30 asymptomatic aPL carriers; 5) 100 healthy donors. Anti-β2GPI-D1 and -D4/5 IgG were tested on research ELISAs containing recombinant β2GPI domains. As compared to other groups, aPL carriers displayed higher frequency/titer of anti-D4/5 IgG. At variance of anti-D4/5, anti-D1 IgG were more frequent and at higher titer in triple than in single/double aPL positive subjects. Anti-D1:anti-D4/5 ratio was predictive of systemic autoimmunity when equal or above 1.5 (OR 3.25, 95% CI 1.45-5.36, p=0.005). Neither anti-D1 nor anti-D4/5 antibodies were associated with APS clinical criteria. Anti-D1 IgG is the preferential specificity not only in vascular and obstetric PAPS, but also in SARD patients with no clinical features of APS. Conversely, aPL carriers do not have a polarized profile toward D1. Combined testing for anti-β2GPI IgG with different domain specificity allows a more accurate aPL profiling, being a polarization toward anti-D1 IgG a possible fingerprint of systemic autoimmunity. This article is protected by copyright. All rights reserved.

Mitochondria morphology in mutant TARDBP and C9ORF72 fibroblasts after 48 h in galactose medium. ... more Mitochondria morphology in mutant TARDBP and C9ORF72 fibroblasts after 48 h in galactose medium. Representative confocal images of mitochondrial network in primary fibroblasts from three healthy controls (C1, C2, C3), three ALS patients carrying mutation in TARDBP gene (p.A382T; T1, T2, T3) and three ALS//FTD patients with pathological expansion of the hexanucleotide repeat in C9ORF72 (C9.2, C9.3, C9.4) transfected with pDsRed2Mito construct. Bar, 10μm. Figure S2. Mitochondria functionality in mutant TARDBP and C9ORF72 fibroblasts. Measurement of mitochondrial membrane potential by flow cytometry analysis of Mitotracker Red (MTR) positive fibroblasts (from 4 healthy controls, 3 TARDBP and 4 C9ORF72 mutated patients) maintained in medium with galactose and without glucose. Median ± SEM, n = 3 different DIV; One-way ANOVA with Dunnett's multiple comparison test; **p

International Journal of Molecular Sciences, 2021

Medullary thyroid carcinoma (MTC) is a tumor deriving from the thyroid C cells. Vandetanib (VAN) ... more Medullary thyroid carcinoma (MTC) is a tumor deriving from the thyroid C cells. Vandetanib (VAN) and cabozantinib (CAB) are two tyrosine kinase inhibitors targeting REarranged during Transfection (RET) and other kinase receptors and are approved for the treatment of advanced MTC. We aim to compare the in vitro and in vivo anti-tumor activity of VAN and CAB in MTC. The effects of VAN and CAB on viability, cell cycle, and apoptosis of TT and MZ-CRC-1 cells are evaluated in vitro using an MTT assay, DNA flow cytometry with propidium iodide, and Annexin V-FITC/propidium iodide staining, respectively. In vivo, the anti-angiogenic potential of VAN and CAB is evaluated in Tg(fli1a:EGFP)y1 transgenic fluorescent zebrafish embryos by analyzing the effects on the physiological development of the sub-intestinal vein plexus and the tumor-induced angiogenesis after TT and MZ-CRC-1 xenotransplantation. VAN and CAB exert comparable effects on TT and MZ-CRC-1 viability inhibition and cell cycle per...

Journal of Translational Autoimmunity, 2021

Arthritis Research & Therapy, 2020

Background Consistently with their diagnostic and prognostic value, autoantibodies specific for s... more Background Consistently with their diagnostic and prognostic value, autoantibodies specific for systemic sclerosis (SSc) embedded in immune complexes (ICs) elicited a pro-inflammatory and pro-fibrotic cascade in healthy skin fibroblasts, engaging Toll-like receptors (TLRs) via their nucleic acid components. The objective of this study was to investigate the pathogenicity of SSc-ICs in endothelial cells. Methods ICs were purified from the sera of SSc patients bearing different autoantibody specificities (antibodies against DNA topoisomerase I, centromeric proteins, RNA polymerase, and Th/To), patients with systemic lupus erythematosus (SLE) and primary anti-phospholipid syndrome (PAPS), or healthy controls (NHS) using polyethylene glycol precipitation. Human umbilical vein endothelial cells (HUVECs) were incubated with ICs, positive and negative controls. mRNA levels of endothelin-1 (et-1), collagenIα1 (colIα1), interferon (IFN)-α, and IFN-β were investigated by real-time PCR; et-1 a...

The Israel Medical Association journal : IMAJ, 2002

Systemic sclerosis is a disease characterized by fibrosis of skin and internal organs. Although a... more Systemic sclerosis is a disease characterized by fibrosis of skin and internal organs. Although at present the classical criteria defining an autoimmune disease are not entirely fulfilled for SSc, several autoimmune abnormalities, both cellular and humoral, have been ...

Arthritis & Rheumatology, 2015

Objective: Rheumatoid arthritis (RA) is associated with accelerated atherosclerosis. The reductio... more Objective: Rheumatoid arthritis (RA) is associated with accelerated atherosclerosis. The reduction of cardiovascular risk induced by methotrexate and anti-TNFα agents in RA is considered secondary to their anti-inflammatory action, but their effects on serum lipoprotein function and foam cell formation are unknown. The reduced high density lipoprotein (HDL) capacity to promote cell cholesterol efflux (CEC) and the increased serum cell cholesterol-loading capacity (CLC) demonstrated in RA may contribute to foam cell development. We investigated the influence of methotrexate and adalimumab treatment on serum CEC and CLC in RA patients. Furthermore, we studied the in vitro effect of the two drugs on macrophage cholesterol handling. Methods: serum from RA patients treated with methotrexate (n=34) or with adalimumab+methotrexate (n=22), drawn before and after 6 weeks and 6 months of treatment, was analyzed for CEC and CLC by radioisotopic and fluorimetric techniques, respectively. Methotrexate and adalimumab influence on macrophage cholesterol efflux and uptake was evaluated in vitro on human THP-1-derived macrophages. Results: methotrexate treatment was associated with serum HDL, LDL and total cholesterol increase, with ATP-binding cassette G1 and Scavenger receptor-BI (SR-BI)-mediated CEC increase and no CLC modification. Adalimumab treatment was associated with serum HDL increase, SR-BI-mediated and ATP-binding cassette A1-mediated CEC transient increase and decrease respectively, and with a significant CLC reduction; in addition, adalimumab reduced macrophage cholesterol uptake in vitro. Conclusion: methotrexate and adalimumab anti-atherosclerotic activity may be mediated by beneficial and complementary effects on lipoprotein functions and on macrophage cholesterol handling, as a whole opposing foam cell formation. This article is protected by copyright. All rights reserved.

Annals of the rheumatic diseases, 2014

The marked cardiovascular risk in autoimmune diseases is only partly explained. The capacity of h... more The marked cardiovascular risk in autoimmune diseases is only partly explained. The capacity of high-density lipoproteins (HDL) to promote cell cholesterol efflux is a property with a well-known anti-atherogenic significance, but is also involved in functional modulation of endothelial and immune cells. The aim of this work was to evaluate HDL functionality with respect to cell cholesterol efflux in rheumatoid arthritis (RA) and systemic lupus erythemathosus (SLE) patients. We evaluated serum cholesterol efflux capacity (CEC) of apoB-depleted serum, which mainly reflects HDL activity, from 30 RA and 30 SLE patients, and from 30 healthy controls by radioisotopic ex-vivo systems discriminating between the specific pathways of cholesterol efflux. RA patients presented impairment of ATP-binding cassette G1-mediated CEC that correlated with disease activity. SLE patients showed a more complex pattern of modifications unrelated to disease activity, with marked reduction of ATP-binding cas...

Trends in Immunology, 2005

Despite the discovery of anti-endothelial cell antibodies (AECA) in a heterogeneous group of diso... more Despite the discovery of anti-endothelial cell antibodies (AECA) in a heterogeneous group of disorders characterized by endothelial damage, their pathogenic role is still debated. Experimental in vitro models indicate that they can either damage endothelial cells or trigger cell signaling by reacting with as yet undefined surface molecules. However, clinical studies suggest that, in addition to AECA, other pathogenic mechanisms are involved in the vasculitic process. Recently, antibodies specific for b2 glycoprotein I, the phospholipid-binding protein targeted by anti-phospholipid antibodies, have been shown to display anti-endothelial activity. These autoantibodies recognize b2 glycoprotein I adhered to the endothelium and induce a cell perturbation that might underlie the thrombophilic state of the antiphospholipid syndrome.

Lupus, 2007

The administration of immunosuppressive drugs during pregnancy is often necessary in women with a... more The administration of immunosuppressive drugs during pregnancy is often necessary in women with autoimmune diseases. Teratogenicity of immunosuppressives during pregnancy has been evaluated, only few data exist about the effects on immune systems. We therefore performed a pilot study on the influence of foetal exposure to immunosuppressives on immune function of babies born to mothers with autoimmune disorders. We investigated serological and cellular parameters as indicators of immune system status. We included in the study 14 babies (mean age 11 months, range 1—24) born to mothers with autoimmune diseases and exposed in utero to different immunosuppressants and, as controls, 14 babies whose mothers had autoimmune manifestations but did not receive immunosuppressive therapy. We evaluated: (i) complete blood count, (ii) immunoglobulin levels and IgG subclasses, (iii) antibody response to hepatitis B vaccine, (iv) leukocyte subpopulations and (v) interleukin-2 and interferon γ in vit...

Journal of Cellular and Molecular Medicine, 2012

Nature reviews. Rheumatology, 2018

Recurrent thrombosis and miscarriages are the main clinical manifestations of antiphospholipid sy... more Recurrent thrombosis and miscarriages are the main clinical manifestations of antiphospholipid syndrome (APS). Although most patients display both clinical signs, some patients can have isolated vascular or obstetric variants. Emerging data raise the question of whether obstetric and vascular APS are the same or different diseases. An important difference between the two conditions is that a thrombophilic state is a common feature in vascular APS, whereas clot occlusions of the decidual spiral arteries are seldom observed in obstetric APS, and infarctions are found in only one-third of APS placentae. Conversely, inflammation, which is undetectable in vascular APS, is frequently observed in the placentae of patients with obstetric APS and has been documented in the placentae of pregnant mice with fetal loss mediated by antiphospholipid antibodies. Attempts to identify different antibodies or epitopes responsible for the two clinical manifestations of APS have so far been unsuccessful...

PloS one, 2017

Domain I (DI) of beta-2-glycoprotein I (β2GPI) contains the immunodominant epitope for pathogenic... more Domain I (DI) of beta-2-glycoprotein I (β2GPI) contains the immunodominant epitope for pathogenic antiphospholipid antibodies (aPL). DI is exposed in the linear form of the molecule but not in the circular form that comprises 90% of serum β2GPI. The majority of circulating β2GPI is biochemically reduced with two free thiols in Domain V. However, increased levels of oxidised β2GPI are found in patients with antiphospholipid syndrome (APS). It is not known whether oxidation of β2GPI favours the linear form of the molecule and thus promotes development of anti-DI antibodies. We investigated whether the proportion of oxidised β2GPI associates with the presence of anti-DI in APS patients. Serum samples from 44 APS patients were screened for IgG, IgM and IgA anti-DI, anti-β2GPI, anti-cardiolipin (anti-CL) and biochemically reduced β2GPI. A negative correlation was found between the proportion of β2GPI in the biochemically reduced form and IgG anti-DI levels (r = -0.54, p = 0.0002), but no...

Microvascular research, 2017

Raynaud's phenomenon (RP) can be the first manifestation of systemic sclerosis (SSc) or other... more Raynaud's phenomenon (RP) can be the first manifestation of systemic sclerosis (SSc) or other connective tissue diseases (CTDs), often preceding an overt disease by years. It is not known if markers of endothelial damage are detectable in those RP patients who subsequently develop a CTD. We studied 82 RP patients at their first evaluation to correlate the levels of endothelial markers with the subsequent development of an overt disease 36months later. We measured plasma levels of tissue-type plasminogen activator (t-PA) and von Willebrand factor (vWF), two markers of endothelial damage, and interleukin-6 (IL-6), a pro-inflammatory cytokine. Thirty sex- and age-matched healthy subjects (HS) served as controls. At baseline, 67 patients showed capillaroscopic normal pattern (CNP) and 15 patients, of which 11 were very early SSc, had capillaroscopic scleroderma pattern (CSP). Plasma levels of t-PA, vWF and IL-6 were higher in patients with CNP (p=0.0001) than in HS and even much hig...

Molecular oncology, Jan 28, 2017

Medullary thyroid cancer (MTC) is a tumor highly resistant to chemo- and radiotherapy. Drug resis... more Medullary thyroid cancer (MTC) is a tumor highly resistant to chemo- and radiotherapy. Drug resistance can be induced by epigenetic changes such as aberrant DNA methylation. To overcome drug resistance, we explored a promising approach based on the use of 5-aza-2'-deoxycytidine (AZA), a demethylating agent, in combination with the mTOR inhibitor everolimus in MTC cells (MZ-CRC-1 and TT). This combined treatment showed a strong synergistic antiproliferative activity through the induction of apoptosis. The effect of everolimus and/or AZA on genome-wide expression profiling was evaluated by Illumina BeadChip in MZ-CRC-1 cells. An innovative bioinformatic pipeline identified four potential molecular pathways implicated in the synergy between AZA and everolimus: PI3K-Akt signaling, the neurotrophin pathway, ECM-receptor interaction and focal adhesion. Among these, the neurotrophin signaling pathway was most directly involved in apoptosis, through the overexpression of NGFR and Bax ge...

The Journal of Immunology, 2017

Journal of acquired immune deficiency syndromes

Sera from patients affected either by the classic/Mediterranean form (5 patients) and human immun... more Sera from patients affected either by the classic/Mediterranean form (5 patients) and human immune deficiency virus (HIV)-associated (12 patients) form of Kaposi's sarcoma (KS) were compared in supporting endothelial cell in-vitro proliferation. Healthy blood donors (15) and a group of 7 HIV-positive drug addicts with no dermatological involvement were used as control groups. Endothelial cell growth was assessed by means of a [3H]thymidine incorporation and by an optical density direct cell-counting assay. Our results indicate that, at the highest concentrations, sera obtained from patients affected by KS with no sign of HIV infection induced significantly higher levels of endothelial cell proliferation when compared with HIV-associated KS and with normal controls. This growth-promoting activity was apparently cell selective, being present in endothelial cell but not in fibroblast cultures.

Journal of clinical & laboratory immunology

The present report investigates the synergistic effects of different agents (phytohaemoagglutinin... more The present report investigates the synergistic effects of different agents (phytohaemoagglutinin (PHA), anti-CD3, phorbol-12-myristate-13-acetate (PMA) and Ionomycin on T cell activation. Results indicate that in normal lymphocytes PMA causes a dose-dependent decrease of PHA-induced blastogenesis and a marked enhancement of anti-CD3-induced 3H-thymidine uptake. There is positive synergism when cells are costimulated with PMA and the calcium ionophore Ionomycin. These data show that phorbol esters can exert either positive or negative synergisms depending on the experimental conditions. We also studied the simultaneous stimulation with PHA and anti-CD3 which gives an addictive effect for the minimal doses of the two stimuli, and an appreciable negative synergism for their highest concentrations. In order to evaluate T lymphocyte functions in the age-associated immune defect, PHA plus anti-CD3 and PMA plus Ionomycin costimulation assays were applied to a population of healthy elderly subjects. An additive effect is found after costimulation with the highest doses of PHA and anti-CD3, at variance with the negative synergism found in younger adult controls. Following costimulation with PMA plus Ionomycin, elderly subjects also display an impaired response, as compared to adult controls. No correlation was found with PHA-, anti-CD3- and PMA plus Ionomycin-induced T cell proliferation. These results underline the complexity of age-associated immune defects and suggest that alterations in different mechanisms of lymphocyte activation are responsible for the immune deficiency of aging.

Infection

The biological response modifying activities of cefodizime (CDZ), a new third-generation cephalos... more The biological response modifying activities of cefodizime (CDZ), a new third-generation cephalosporin, were investigated in vitro and ex vivo. In vitro investigations using cells isolated from the blood of young healthy donors showed no stimulating activity of CDZ on peripheral blood lymphocytes, natural killer cell activity, IL-1 production by adherent mononuclear cells, PMN chemiluminescence or PMN chemotaxis. A slight but statistically insignificant increase in PMN phagocytosis and phagocytic index was observed in the same population. IL-1 production was increased in three subjects with low resting state values. In a controlled ex vivo study, 20 healthy elderly subjects selected on the basis of depressed phagocytic function were treated with CDZ 1 g i.m. b.i.d. or placebo for eight days. PMN function was determined at baseline and on the day after the last dose. In the CDZ group a significant increase in both phagocytosis and phagocytic index was found, while there were no chang...

Annals of the Rheumatic Diseases, 2014

ABSTRACT Background Raynaud's phenomenon (RP) may predate an overt Connective Tissue Dise... more ABSTRACT Background Raynaud's phenomenon (RP) may predate an overt Connective Tissue Disease (CTD) of the “Scleroderma spectrum” such as Systemic sclerosis (SSc). The predictive role of auto-antibodies (auto-Abs) such as ANA for the development of a CTD in patients with isolated RP (i.e. with no other signs or symptoms of CTD) is well known, whereas data regarding the prevalence of anti-ENA in such patients are lacking. Objectives Our aim was to evaluate the profile of anti-ENA in patients with isolated RP. Methods Sera were analyzed for anti-ENA by EUROLINE “Scleroderma profile” (antigens: Scl70, CENP A, CENP B, RP11, RP155, fibrillarin (Fib), NOR90, Th/To, PM-Scl100, PM-Scl75, Ku, PDGFR, Ro-52) and “Myosites profile” (antigens: Mi-2, Ku, PM-Scl100, PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ, Ro-52) [EUROIMMUN AG Leuback, Germany]. Results A total of 115 adults (105 females and 10 males) with isolated RP were enrolled in two Italian Rheumatology Centers (G. Pini Institute, Milan and Moriggia-Pelascini Hospital, Gravedona) from March 2011 to September 2013. Forty-four of 115 (38.0%) subjects were positive for at least one anti-ENA; 71 out of 110 (61.7%) were negative both for Scleroderma and Myosites profiles (see table 1 and 2). Conclusions Up to 38% of RP subjects were positive for at least one anti-ENA. The detection of anti-ENA since the first Rheumatologic evaluation may aid the specialist in achieving an earlier diagnosis. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5955

Arthritis & Rheumatology, 2015

It was suggested that only antibodies against Domain 1 (D1) of β2glycoprotein I (β2 GPI) are path... more It was suggested that only antibodies against Domain 1 (D1) of β2glycoprotein I (β2 GPI) are pathogenic and diagnostic. Still debated is the role of antibodies against other β2GPI domains. Our aim was to evaluate the clinical relevance of domain specificity profiling of anti-β2GPI IgG antibodies not only in Antiphospholipid Syndrome (APS) patients but also in control groups of Systemic Autoimmune Rheumatic Diseases (SARDs) and asymptomatic Antiphospholipid Antibody (aPL) carriers. 159 subjects with persistently positive, medium/high titer anti-β2GPI IgG were included: 1) 56 thrombotic (+/- obstetric) primary APS (PAPS); 2) 31 women with pure obstetric PAPS; 3) 42 aPL-positive SARDs; 4) 30 asymptomatic aPL carriers; 5) 100 healthy donors. Anti-β2GPI-D1 and -D4/5 IgG were tested on research ELISAs containing recombinant β2GPI domains. As compared to other groups, aPL carriers displayed higher frequency/titer of anti-D4/5 IgG. At variance of anti-D4/5, anti-D1 IgG were more frequent and at higher titer in triple than in single/double aPL positive subjects. Anti-D1:anti-D4/5 ratio was predictive of systemic autoimmunity when equal or above 1.5 (OR 3.25, 95% CI 1.45-5.36, p=0.005). Neither anti-D1 nor anti-D4/5 antibodies were associated with APS clinical criteria. Anti-D1 IgG is the preferential specificity not only in vascular and obstetric PAPS, but also in SARD patients with no clinical features of APS. Conversely, aPL carriers do not have a polarized profile toward D1. Combined testing for anti-β2GPI IgG with different domain specificity allows a more accurate aPL profiling, being a polarization toward anti-D1 IgG a possible fingerprint of systemic autoimmunity. This article is protected by copyright. All rights reserved.