Marco Cardone - Academia.edu (original) (raw)

Papers by Marco Cardone

Journal of Allergy and Clinical Immunology

The Journal of Immunology

Immune sensing of fungi occurs via multiple pathogen recognition receptors such as Dectin-1/2, ma... more Immune sensing of fungi occurs via multiple pathogen recognition receptors such as Dectin-1/2, mannose receptor and TLR2. The β-glucans and mannans in fungal cell walls activate these receptors. Stimulation of DCs via TLR2 results in the production of IL-10 and little IL-12, while Dectin-1 utilizes a hemi-ITAM (immunoreceptor tyrosine based activation motif) and Syk to facilitate IL-23 production leading to Th17 polarization. We have recently shown that the linker for activation of B cells/Non-T cell activating linker (LAB/NTAL), a known regulator of ITAM-mediated signaling, is expressed in BMDCs. Here we examine whether LAB regulates the response of BMDC to zymosan by affecting Dectin-1 signaling. We find that LAB, Syk, Cbl, MAPK and Akt are rapidly and transiently phosphorylated following zymosan stimulation. Moreover, zymosan stimulated Lat2-/- BMDC produced decreased IL-10 and IL-12p40 and increased TNF levels suggesting the involvement of LAB in regulating BMDC cytokine product...

The Journal of Immunology

We have previously demonstrated in human monocyte-derived dendritic cells (DCs), interleukin-23 (... more We have previously demonstrated in human monocyte-derived dendritic cells (DCs), interleukin-23 (IL-23) is efficiently induced by the Dectin-1 ligands alone or in combination with Toll receptor 2/6 (TLR2/6) ligand Pam2C but not by the TLR4 ligand LPS alone. We found that the ability of various stimuli to induce Th17 cells depends not only on their induction of IL-23, IL-6, and TGF-β production in DCs, but also on their ability to induce bioactive IL-1β. We considered whether the Dectin-1 receptor ligands were able directly or indirectly to activate the inflammasome. We found that only particulate but not soluble forms of β-glucan induced secretion of IL-1β in human DCs, and caspase-1 inhibitors blocked its secretion. LPS did not induce IL-1β secretion unless ATP was added to activate the inflammasome. Moreover, treatment with oxidized ATP or apyrase impaired the secretion of IL-1β by β-glucan-treated DCs. We also observed that caspase-1 inhibitors or an IL-1 receptor antagonist sign...

The Journal of Immunology

The molecular networks controlling the DC programming in response to C-type lectin receptor agoni... more The molecular networks controlling the DC programming in response to C-type lectin receptor agonists including beta-glucan, a fungal and bacterial component and ligand for the Dectin-1 receptor, are still poorly characterized. We studied these immunological aspects by comparing the response of human mono-DCs to the Dectin-1 ligand, beta-glucan, and to the TLR4 ligand, LPS using a gene expression/perturbation approach. Gene expression analysis of mono-DCs stimulated by beta-glucan identified IL-1, TNF, and type I IFN encoded by “early/immediate” type genes as predicted regulators of the beta-glucan-induced “late” transcriptional response. A perturbation analysis revealed that endogenous IL-1, via the nuclear factor IkappaB-zeta, selectively promoted the expression of beta-glucan-induced “late” genes such as those encoding IL-6 and IL-23, thus programming DCs to initiate Th17 responses. TNF and type I IFN were found to modulate the response to both beta-glucan and LPS. IFN-gamma count...

The Journal of Immunology

Adverse drug reactions (ADR) are a major obstacle to drug development. Genome-wide association st... more Adverse drug reactions (ADR) are a major obstacle to drug development. Genome-wide association studies identified several human leukocyte antigens (HLA)-class I alleles as risk factors for ADR. The HLA-B*57:01 allele has been found to be associated with the development of abacavir (ABC) hypersensitivity syndrome (AHS) and with the induction of liver injury by the β-lactam antibiotic flucloxacillin. The nucleoside analog ABC, an inhibitor of the HIV reverse transcriptase, can induce severe multi-organ toxicity in >50% of HLA-B*57:01+ patients with HIV infection. In a previous study we showed that ABC induced binding to the HLA-B*57:01 of altered self-peptides containing predominantly isoleucine or leucine residues at the carboxyl terminus. Recognition of these self-peptides drives in vitro activation of cytotoxic CD8+ T cells. However, the early immune events/danger signals required to overcome the immune tolerance that otherwise suppress ADR are still unknown. In order to study H...

The Journal of Immunology

Signaling through the adaptor protein MyD88 promotes carcinogenesis in several chemically induced... more Signaling through the adaptor protein MyD88 promotes carcinogenesis in several chemically induced cancer models. In contrast, in the AOM/DSS model of chronic colitis induced cancer, we observed a protective role for MyD88 in the development of colitis-associated cancer (CAC). The inability of Myd88-/- mice to heal ulcers generated upon DSS treatment creates an altered inflammatory environment that induces profound early alterations in expression of genes encoding pro-inflammatory factors as well as pathways regulating cell proliferation and apoptosis. Additionally, AOM/DSS treatment induced alterations in DNA repair in MyD88 deficient mice, resulting in increased frequency of mutations and a dramatic increase in adenoma formation and progression to invasive phenotype. Others have reported that Tlr4 deficient mice share similar susceptibility to colitis as Myd88 deficient mice but, unlike the latter, are resistant to CAC. We observed that mice deficient for Tlr2 or IL-1R do not show ...

Journal of Immunology, May 1, 2014

Triggering Receptor Expressed on Myeloid cells (TREM) family receptors modulate responses of a va... more Triggering Receptor Expressed on Myeloid cells (TREM) family receptors modulate responses of a variety of leukocytes. We found that TREM-like 4 (TREML4) mRNA is elevated in the peripheral blood of patients with advanced coronary artery calcification (CAC) and that a single nucleotide polymorphism (SNP), rs2803496, tracks with the level of TREML4 mRNA in these patients. In an effort to elucidate the mechanism through which TREML4 in peripheral blood could affect CAC, we began by determining what cells express it. Monocytes and lymphocytes of donors with this SNP expressed very low levels of TREML4 mRNA whereas their neutrophils (PMNs) were strongly positive. Consistent with its association with CAC, we found variation in the levels of TREML4 mRNA in PMNs that correlated rs2803496, and a second SNP rs2803495. Unlike murine Treml4, which has the typical TREM family structure, human TREML4 has a noncanonical leader, lacks a portion of the transmembrane domain (TM) and has no cytoplasmic tail. Characterization of the Treml4 cDNA suggests that this truncated TM domain is sufficient to anchor the protein in the plasma membrane but that the TREML4 native leader sequence is ineffective. Lastly, immunostaining of human atherosclerotic plaques shows TREML4 in macrophage-rich regions, as well as on the endothelium. Together our findings suggest that TREML4 may play an important role in atherosclerosis and CAC but that it is unlikely to function as a typical, DAP12-coupled TREM.

Frontiers in Immunology, 2021

Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-... more Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-peptide complexes can lead to severe hypersensitivity reactions. Flucloxacillin (FLX), a β-lactam antibiotic for narrow-spectrum gram-positive bacterial infections, has been associated with severe immune-mediated drug-induced liver injury caused by an influx of T-lymphocytes targeting liver cells potentially recognizing drug-haptenated peptides in the context of HLA-B*57:01. To identify immunopeptidome changes that could lead to drug-driven immunogenicity, we used mass spectrometry to characterize the proteome and immunopeptidome of B-lymphoblastoid cells solely expressing HLA-B*57:01 as MHC-I molecules. Selected drug-conjugated peptides identified in these cells were synthesized and tested for their immunogenicity in HLA-B*57:01-transgenic mice. T cell responses were evaluated in vitro by immune assays. The immunopeptidome of FLX-treated cells was more diverse than that of untreated cel...

Archives of Medical Research, 2018

Background. The diuretic response has been shown to be a robust independent marker of cardiovascu... more Background. The diuretic response has been shown to be a robust independent marker of cardiovascular outcomes in acute heart failure (ADHF) patients. The objectives of this clinical research, will aim are to: a) include diuresis in the formula for diuretic response (R-to-D); b) add to R-to-D the value of a pre-discharged determination of galectin-3 and BNP in predicting mid-term clinical outcome. Methods. Consecutive patients discharged alive after an ADHF were enrolled. All patients underwent BNP and galectin-3, a 6 min walk test and an echocardiogram together with diuresis and body weight during diuretic administration. Death by any cause, cardiac transplantation and worsening HF requiring readmission to the hospital were considered cardiovascular events. Results. 141 patients (98 males, age 73.8) were analysed (follow-up 17 months). During the follow-up 45 (31.9%) events were scheduled (19 cardiac deaths, 26 re-hospitalisation for HF). Patients who experienced CV-event had a worst renal function (p 5 0.003), an higher BNP (p 5 0.006) and galectin-3 (p 5 0.008). At multivariate analysis, only R-to-D, galectin-3 and BNP showed a significant correlation with worst clinical prognosis (respectively p 5 0.043; OR 6.01; p 5 0.01; OR 8.9; p 5 0.02 OR 10.38), independently of age and renal function. Kaplan-Meier curves depicted a powerful stratification using an R-to-D !1.2 kg/40 mg furosemide (log rank 10.96; p 5 0.0009). Adding R-to-D! 1.2 mg/40 mg furosemide to galectin-3O17.6 pg/mL and BNPO500 pg/mL the predictive value improved (log rank 23.59; p 5 0.0001). Conclusion. Adding R-to-D to Gal-3 and BNP, a single pre-discharge strategy testing seemed to obtain a satisfactorily predictive value in alive HF patients discharged after an ADHF episode.

Frontiers in Immunology, 2020

The first case of human transmission of SARS-CoV-2 was reported in China in December 2019. A few ... more The first case of human transmission of SARS-CoV-2 was reported in China in December 2019. A few months later, this viral infection had spread worldwide and became a pandemic. The disease caused by SARS-CoV-2, termed COVID-19, is multifactorial and associated with both specific antiviral as well as inflammatory responses, the extent of which may determine why some individuals are asymptomatic while others develop serious complications. Here we review possible life-threating immune events that can occur during disease progression to uncover key factors behind COVID-19 severity and provide suggestions for interventions with repurposed drugs in wellcontrolled and randomized clinical trials. These drugs include therapeutics with potential to inhibit SARS-CoV-2 entry into host cells such as serine protease inhibitors of the cellular protease TMPS2 and drugs targeting the renin-angiotensin system; antivirals with potential to block SARS-CoV-2 replication or factors that could boost the antiviral response; monoclonal antibodies targeting pro-inflammatory cytokines that drive the hyperinflammatory response during COVID-19 progression toward the severe stage and therapeutics that could ameliorate the function of the lungs. Furthermore, in order to help make more informed decisions on the timing of the intervention with the drugs listed in this review, we have grouped these therapeutics according to the stage of COVID-19 progression that we considered most appropriate for their mechanism of action.

The Journal of clinical investigation, Jan 21, 2018

Adverse drug reactions (ADRs) are a major obstacle to drug development, and some of these, includ... more Adverse drug reactions (ADRs) are a major obstacle to drug development, and some of these, including hypersensitivity reactions to the HIV reverse transcriptase inhibitor abacavir (ABC), are associated with HLA alleles, particularly HLA-B*57:01. However, not all HLA-B*57:01+ patients develop ADRs, suggesting that in addition to the HLA genetic risk, other factors may influence the outcome of the response to the drug. To study HLA-linked ADRs in vivo, we generated HLA-B*57:01-Tg mice and show that, although ABC activated Tg mouse CD8+ T cells in vitro in a HLA-B*57:01-dependent manner, the drug was tolerated in vivo. In immunocompetent Tg animals, ABC induced CD8+ T cells with an anergy-like phenotype that did not lead to ADRs. In contrast, in vivo depletion of CD4+ T cells prior to ABC administration enhanced DC maturation to induce systemic ABC-reactive CD8+ T cells with an effector-like and skin-homing phenotype along with CD8+ infiltration and inflammation in drug-sensitized skin...

Journal of Theoretical and Applied Vascular Research, 2016

Lymphedema in many countries is still considered an "aesthetic" problem, even though a chronic, d... more Lymphedema in many countries is still considered an "aesthetic" problem, even though a chronic, degenerative, and debilitating disease. After a long period of study with internal and external experts, the Italian Ministry of Health has implemented National Care guidelines for lymphedema and other related disorders. This Document represents a fundamental element with regards to diagnostic and therapeutic regulation procedures that satisfy the health needs of these patients. In this paper the description of the main constituents of the document that has revolutionized the possibility of access the best specialized care facilities situated in the country.

Medicine, 2016

Galectin-3 demonstrated to be a robust independent marker of cardiovascular mid-term (18-month) o... more Galectin-3 demonstrated to be a robust independent marker of cardiovascular mid-term (18-month) outcome in heart failure (HF) patients. The objective of this study was to analyze the value of a predischarged determination of plasma galectin-3 alone and with plasma brain natriuretic peptide (BNP) in predicting mid-term outcome in frequent-flyers (FF) HF (≥2 hospitalization for HF/year)/dead patients discharged after an acute decompensated HF (ADHF) episode. All FF chronic HF subjects discharged alive after an ADHF were enrolled. All patients underwent a determination of BNP and galectin-3, a 6-minute walk test, and an echocardiogram within 48 hours upon hospital discharge. Death by any cause, cardiac transplantation, and worsening HF requiring readmission to hospital were considered cardiovascular events. Eighty-three patients (67 males, age 73.2 ± 8.6 years old) were analyzed (mean follow-up 11.6 ± 5.2 months; range 4-22 months). During the follow-up 38 events (45.7%) were scheduled: (13 cardiac deaths, 35 rehospitalizations for ADHF). According to medical history, in 33 patients (39.8%) a definition of FF HF patients was performed (range 2-4 hospitalization/year). HF patients who suffered an event (FF or death) demonstrated more impaired ventricular function (P = 0.037), higher plasma BNP (P = 0.005), and Gal-3 at predischarge evaluation (P = 0.027). Choosing adequate cutoff points (BNP ≥ 500 pg/mL and Gal-3 ≥ 17.6 ng/mL), the Kaplan-Meier curves depicted the powerful stratification using BNP + Gal-3 in predicting clinical course at mid-term follow-up (log rank 5.65; P = 0.017). Adding Gal-3 to BNP, a single predischarge strategy testing seemed to obtain a satisfactorily predictive value in alive HF patients discharged after an ADHF episode. Abbreviations: ADHF = acute decompensated heart failure, BNP = brain natriuretic peptide, FF = frequent-flyers, HF = heart failure, LOS = length of hospital stay, LVEF = left ventricle ejection fraction, NYHA = New York Heart Association.

Nucleic acids research, Jan 24, 2015

Oligonucleotide aptamers represent a novel platform for creating ligands with desired specificity... more Oligonucleotide aptamers represent a novel platform for creating ligands with desired specificity, and they offer many potentially significant advantages over monoclonal antibodies in terms of feasibility, cost, and clinical applicability. However, the isolation of high-affinity aptamer ligands from random oligonucleotide pools has been challenging. Although high-throughput sequencing (HTS) promises to significantly facilitate systematic evolution of ligands by exponential enrichment (SELEX) analysis, the enormous datasets generated in the process pose new challenges for identifying those rare, high-affinity aptamers present in a given pool. We show that emulsion PCR preserves library diversity, preventing the loss of rare high-affinity aptamers that are difficult to amplify. We also demonstrate the importance of using reference targets to eliminate binding candidates with reduced specificity. Using a combination of bioinformatics and functional analyses, we show that the rate of am...

Hemangiomas and Vascular Malformations, 2015

The phlebolymphedema of the limbs is a disorder with frequent occurrence in its various forms, wh... more The phlebolymphedema of the limbs is a disorder with frequent occurrence in its various forms, which by its very nature tends to worsen and has a tendency to complications and clinical exacerbation.

Veins and Lymphatics, 2012

Hyperhomocysteinemia is a widely recognized, although not yet entirely understood, risk factor fo... more Hyperhomocysteinemia is a widely recognized, although not yet entirely understood, risk factor for cardiovascular disease. Particularly, the complex relationships between age, hyperhomocysteinemia, predisposing genetic factors and peripheral vascular diseases have not been fully evaluated. Our contribution to this issue is a retrospective analysis of a large series of patients with peripheral arterial, venous and lymphatic disease, and of their blood relatives, with special reference to homocysteine plasma levels, age and methylenetetrahydrofolate reductase (MTHFR) polymorphisms. Serum homocysteine was measured in 477 patients (286 males, 191 females, age range 19-78 years) with various vascular clinical conditions: postphlebitic syndrome (46) recurrent venous ulcers (78), arterial diseases (101) primary lymphoedema (87), secondary lymphoedema (161) and outlet thoracic syndrome (4), and in 50 normal controls. A MTHFR study for polymorphisms was carried on in the subjects with homocysteine values exceeding 15 mol/L. Serum homocysteine determination and MTHFR polymorphism studies were performed also in 1430 healthy blood related relatives (mainly siblings, descendents and sibling descendents) of the subjects with hyperhomocysteinemia and MTHFR polymorphisms. We found MTHFR polymorphisms in 20% of controls and in 69.3%, 69.5% and 53.8% of hyperhomocysteinemic subjects with arterial diseases, postphlebitic syndrome and venous ulcers, respectively. As expected, the percentage of hyperhomocysteinemia in patients with secondary lym

PLoS ONE, 2014

Recognition of microbial components via innate receptors including the C-type lectin receptor Dec... more Recognition of microbial components via innate receptors including the C-type lectin receptor Dectin-1, together with the inflammatory environment, programs dendritic cells (DCs) to orchestrate the magnitude and type of adaptive immune responses. The exposure to b-glucan, a known Dectin-1 agonist and component of fungi, yeasts, and certain immune support supplements, activates DCs to induce T helper (Th)17 cells that are essential against fungal pathogens and extracellular bacteria but may trigger inflammatory pathology or autoimmune diseases. However, the exact mechanisms of DC programming by b-glucan have not yet been fully elucidated. Using a gene expression/perturbation approach, we demonstrate that in human DCs b-glucan transcriptionally activates via an interleukin (IL)-1-and inflammasome-mediated positive feedback late-induced genes that bridge innate and adaptive immunity. We report that in addition to its known ability to directly prime T cells toward the Th17 lineage, IL-1 by promoting the transcriptional cofactor inhibitor of kB-f (IkB-f) also programs b-glucan-exposed

Journal of Allergy and Clinical Immunology

The Journal of Immunology

Immune sensing of fungi occurs via multiple pathogen recognition receptors such as Dectin-1/2, ma... more Immune sensing of fungi occurs via multiple pathogen recognition receptors such as Dectin-1/2, mannose receptor and TLR2. The β-glucans and mannans in fungal cell walls activate these receptors. Stimulation of DCs via TLR2 results in the production of IL-10 and little IL-12, while Dectin-1 utilizes a hemi-ITAM (immunoreceptor tyrosine based activation motif) and Syk to facilitate IL-23 production leading to Th17 polarization. We have recently shown that the linker for activation of B cells/Non-T cell activating linker (LAB/NTAL), a known regulator of ITAM-mediated signaling, is expressed in BMDCs. Here we examine whether LAB regulates the response of BMDC to zymosan by affecting Dectin-1 signaling. We find that LAB, Syk, Cbl, MAPK and Akt are rapidly and transiently phosphorylated following zymosan stimulation. Moreover, zymosan stimulated Lat2-/- BMDC produced decreased IL-10 and IL-12p40 and increased TNF levels suggesting the involvement of LAB in regulating BMDC cytokine product...

The Journal of Immunology

We have previously demonstrated in human monocyte-derived dendritic cells (DCs), interleukin-23 (... more We have previously demonstrated in human monocyte-derived dendritic cells (DCs), interleukin-23 (IL-23) is efficiently induced by the Dectin-1 ligands alone or in combination with Toll receptor 2/6 (TLR2/6) ligand Pam2C but not by the TLR4 ligand LPS alone. We found that the ability of various stimuli to induce Th17 cells depends not only on their induction of IL-23, IL-6, and TGF-β production in DCs, but also on their ability to induce bioactive IL-1β. We considered whether the Dectin-1 receptor ligands were able directly or indirectly to activate the inflammasome. We found that only particulate but not soluble forms of β-glucan induced secretion of IL-1β in human DCs, and caspase-1 inhibitors blocked its secretion. LPS did not induce IL-1β secretion unless ATP was added to activate the inflammasome. Moreover, treatment with oxidized ATP or apyrase impaired the secretion of IL-1β by β-glucan-treated DCs. We also observed that caspase-1 inhibitors or an IL-1 receptor antagonist sign...

The Journal of Immunology

The molecular networks controlling the DC programming in response to C-type lectin receptor agoni... more The molecular networks controlling the DC programming in response to C-type lectin receptor agonists including beta-glucan, a fungal and bacterial component and ligand for the Dectin-1 receptor, are still poorly characterized. We studied these immunological aspects by comparing the response of human mono-DCs to the Dectin-1 ligand, beta-glucan, and to the TLR4 ligand, LPS using a gene expression/perturbation approach. Gene expression analysis of mono-DCs stimulated by beta-glucan identified IL-1, TNF, and type I IFN encoded by “early/immediate” type genes as predicted regulators of the beta-glucan-induced “late” transcriptional response. A perturbation analysis revealed that endogenous IL-1, via the nuclear factor IkappaB-zeta, selectively promoted the expression of beta-glucan-induced “late” genes such as those encoding IL-6 and IL-23, thus programming DCs to initiate Th17 responses. TNF and type I IFN were found to modulate the response to both beta-glucan and LPS. IFN-gamma count...

The Journal of Immunology

Adverse drug reactions (ADR) are a major obstacle to drug development. Genome-wide association st... more Adverse drug reactions (ADR) are a major obstacle to drug development. Genome-wide association studies identified several human leukocyte antigens (HLA)-class I alleles as risk factors for ADR. The HLA-B*57:01 allele has been found to be associated with the development of abacavir (ABC) hypersensitivity syndrome (AHS) and with the induction of liver injury by the β-lactam antibiotic flucloxacillin. The nucleoside analog ABC, an inhibitor of the HIV reverse transcriptase, can induce severe multi-organ toxicity in >50% of HLA-B*57:01+ patients with HIV infection. In a previous study we showed that ABC induced binding to the HLA-B*57:01 of altered self-peptides containing predominantly isoleucine or leucine residues at the carboxyl terminus. Recognition of these self-peptides drives in vitro activation of cytotoxic CD8+ T cells. However, the early immune events/danger signals required to overcome the immune tolerance that otherwise suppress ADR are still unknown. In order to study H...

The Journal of Immunology

Signaling through the adaptor protein MyD88 promotes carcinogenesis in several chemically induced... more Signaling through the adaptor protein MyD88 promotes carcinogenesis in several chemically induced cancer models. In contrast, in the AOM/DSS model of chronic colitis induced cancer, we observed a protective role for MyD88 in the development of colitis-associated cancer (CAC). The inability of Myd88-/- mice to heal ulcers generated upon DSS treatment creates an altered inflammatory environment that induces profound early alterations in expression of genes encoding pro-inflammatory factors as well as pathways regulating cell proliferation and apoptosis. Additionally, AOM/DSS treatment induced alterations in DNA repair in MyD88 deficient mice, resulting in increased frequency of mutations and a dramatic increase in adenoma formation and progression to invasive phenotype. Others have reported that Tlr4 deficient mice share similar susceptibility to colitis as Myd88 deficient mice but, unlike the latter, are resistant to CAC. We observed that mice deficient for Tlr2 or IL-1R do not show ...

Journal of Immunology, May 1, 2014

Triggering Receptor Expressed on Myeloid cells (TREM) family receptors modulate responses of a va... more Triggering Receptor Expressed on Myeloid cells (TREM) family receptors modulate responses of a variety of leukocytes. We found that TREM-like 4 (TREML4) mRNA is elevated in the peripheral blood of patients with advanced coronary artery calcification (CAC) and that a single nucleotide polymorphism (SNP), rs2803496, tracks with the level of TREML4 mRNA in these patients. In an effort to elucidate the mechanism through which TREML4 in peripheral blood could affect CAC, we began by determining what cells express it. Monocytes and lymphocytes of donors with this SNP expressed very low levels of TREML4 mRNA whereas their neutrophils (PMNs) were strongly positive. Consistent with its association with CAC, we found variation in the levels of TREML4 mRNA in PMNs that correlated rs2803496, and a second SNP rs2803495. Unlike murine Treml4, which has the typical TREM family structure, human TREML4 has a noncanonical leader, lacks a portion of the transmembrane domain (TM) and has no cytoplasmic tail. Characterization of the Treml4 cDNA suggests that this truncated TM domain is sufficient to anchor the protein in the plasma membrane but that the TREML4 native leader sequence is ineffective. Lastly, immunostaining of human atherosclerotic plaques shows TREML4 in macrophage-rich regions, as well as on the endothelium. Together our findings suggest that TREML4 may play an important role in atherosclerosis and CAC but that it is unlikely to function as a typical, DAP12-coupled TREM.

Frontiers in Immunology, 2021

Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-... more Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-peptide complexes can lead to severe hypersensitivity reactions. Flucloxacillin (FLX), a β-lactam antibiotic for narrow-spectrum gram-positive bacterial infections, has been associated with severe immune-mediated drug-induced liver injury caused by an influx of T-lymphocytes targeting liver cells potentially recognizing drug-haptenated peptides in the context of HLA-B*57:01. To identify immunopeptidome changes that could lead to drug-driven immunogenicity, we used mass spectrometry to characterize the proteome and immunopeptidome of B-lymphoblastoid cells solely expressing HLA-B*57:01 as MHC-I molecules. Selected drug-conjugated peptides identified in these cells were synthesized and tested for their immunogenicity in HLA-B*57:01-transgenic mice. T cell responses were evaluated in vitro by immune assays. The immunopeptidome of FLX-treated cells was more diverse than that of untreated cel...

Archives of Medical Research, 2018

Background. The diuretic response has been shown to be a robust independent marker of cardiovascu... more Background. The diuretic response has been shown to be a robust independent marker of cardiovascular outcomes in acute heart failure (ADHF) patients. The objectives of this clinical research, will aim are to: a) include diuresis in the formula for diuretic response (R-to-D); b) add to R-to-D the value of a pre-discharged determination of galectin-3 and BNP in predicting mid-term clinical outcome. Methods. Consecutive patients discharged alive after an ADHF were enrolled. All patients underwent BNP and galectin-3, a 6 min walk test and an echocardiogram together with diuresis and body weight during diuretic administration. Death by any cause, cardiac transplantation and worsening HF requiring readmission to the hospital were considered cardiovascular events. Results. 141 patients (98 males, age 73.8) were analysed (follow-up 17 months). During the follow-up 45 (31.9%) events were scheduled (19 cardiac deaths, 26 re-hospitalisation for HF). Patients who experienced CV-event had a worst renal function (p 5 0.003), an higher BNP (p 5 0.006) and galectin-3 (p 5 0.008). At multivariate analysis, only R-to-D, galectin-3 and BNP showed a significant correlation with worst clinical prognosis (respectively p 5 0.043; OR 6.01; p 5 0.01; OR 8.9; p 5 0.02 OR 10.38), independently of age and renal function. Kaplan-Meier curves depicted a powerful stratification using an R-to-D !1.2 kg/40 mg furosemide (log rank 10.96; p 5 0.0009). Adding R-to-D! 1.2 mg/40 mg furosemide to galectin-3O17.6 pg/mL and BNPO500 pg/mL the predictive value improved (log rank 23.59; p 5 0.0001). Conclusion. Adding R-to-D to Gal-3 and BNP, a single pre-discharge strategy testing seemed to obtain a satisfactorily predictive value in alive HF patients discharged after an ADHF episode.

Frontiers in Immunology, 2020

The first case of human transmission of SARS-CoV-2 was reported in China in December 2019. A few ... more The first case of human transmission of SARS-CoV-2 was reported in China in December 2019. A few months later, this viral infection had spread worldwide and became a pandemic. The disease caused by SARS-CoV-2, termed COVID-19, is multifactorial and associated with both specific antiviral as well as inflammatory responses, the extent of which may determine why some individuals are asymptomatic while others develop serious complications. Here we review possible life-threating immune events that can occur during disease progression to uncover key factors behind COVID-19 severity and provide suggestions for interventions with repurposed drugs in wellcontrolled and randomized clinical trials. These drugs include therapeutics with potential to inhibit SARS-CoV-2 entry into host cells such as serine protease inhibitors of the cellular protease TMPS2 and drugs targeting the renin-angiotensin system; antivirals with potential to block SARS-CoV-2 replication or factors that could boost the antiviral response; monoclonal antibodies targeting pro-inflammatory cytokines that drive the hyperinflammatory response during COVID-19 progression toward the severe stage and therapeutics that could ameliorate the function of the lungs. Furthermore, in order to help make more informed decisions on the timing of the intervention with the drugs listed in this review, we have grouped these therapeutics according to the stage of COVID-19 progression that we considered most appropriate for their mechanism of action.

The Journal of clinical investigation, Jan 21, 2018

Adverse drug reactions (ADRs) are a major obstacle to drug development, and some of these, includ... more Adverse drug reactions (ADRs) are a major obstacle to drug development, and some of these, including hypersensitivity reactions to the HIV reverse transcriptase inhibitor abacavir (ABC), are associated with HLA alleles, particularly HLA-B*57:01. However, not all HLA-B*57:01+ patients develop ADRs, suggesting that in addition to the HLA genetic risk, other factors may influence the outcome of the response to the drug. To study HLA-linked ADRs in vivo, we generated HLA-B*57:01-Tg mice and show that, although ABC activated Tg mouse CD8+ T cells in vitro in a HLA-B*57:01-dependent manner, the drug was tolerated in vivo. In immunocompetent Tg animals, ABC induced CD8+ T cells with an anergy-like phenotype that did not lead to ADRs. In contrast, in vivo depletion of CD4+ T cells prior to ABC administration enhanced DC maturation to induce systemic ABC-reactive CD8+ T cells with an effector-like and skin-homing phenotype along with CD8+ infiltration and inflammation in drug-sensitized skin...

Journal of Theoretical and Applied Vascular Research, 2016

Lymphedema in many countries is still considered an "aesthetic" problem, even though a chronic, d... more Lymphedema in many countries is still considered an "aesthetic" problem, even though a chronic, degenerative, and debilitating disease. After a long period of study with internal and external experts, the Italian Ministry of Health has implemented National Care guidelines for lymphedema and other related disorders. This Document represents a fundamental element with regards to diagnostic and therapeutic regulation procedures that satisfy the health needs of these patients. In this paper the description of the main constituents of the document that has revolutionized the possibility of access the best specialized care facilities situated in the country.

Medicine, 2016

Galectin-3 demonstrated to be a robust independent marker of cardiovascular mid-term (18-month) o... more Galectin-3 demonstrated to be a robust independent marker of cardiovascular mid-term (18-month) outcome in heart failure (HF) patients. The objective of this study was to analyze the value of a predischarged determination of plasma galectin-3 alone and with plasma brain natriuretic peptide (BNP) in predicting mid-term outcome in frequent-flyers (FF) HF (≥2 hospitalization for HF/year)/dead patients discharged after an acute decompensated HF (ADHF) episode. All FF chronic HF subjects discharged alive after an ADHF were enrolled. All patients underwent a determination of BNP and galectin-3, a 6-minute walk test, and an echocardiogram within 48 hours upon hospital discharge. Death by any cause, cardiac transplantation, and worsening HF requiring readmission to hospital were considered cardiovascular events. Eighty-three patients (67 males, age 73.2 ± 8.6 years old) were analyzed (mean follow-up 11.6 ± 5.2 months; range 4-22 months). During the follow-up 38 events (45.7%) were scheduled: (13 cardiac deaths, 35 rehospitalizations for ADHF). According to medical history, in 33 patients (39.8%) a definition of FF HF patients was performed (range 2-4 hospitalization/year). HF patients who suffered an event (FF or death) demonstrated more impaired ventricular function (P = 0.037), higher plasma BNP (P = 0.005), and Gal-3 at predischarge evaluation (P = 0.027). Choosing adequate cutoff points (BNP ≥ 500 pg/mL and Gal-3 ≥ 17.6 ng/mL), the Kaplan-Meier curves depicted the powerful stratification using BNP + Gal-3 in predicting clinical course at mid-term follow-up (log rank 5.65; P = 0.017). Adding Gal-3 to BNP, a single predischarge strategy testing seemed to obtain a satisfactorily predictive value in alive HF patients discharged after an ADHF episode. Abbreviations: ADHF = acute decompensated heart failure, BNP = brain natriuretic peptide, FF = frequent-flyers, HF = heart failure, LOS = length of hospital stay, LVEF = left ventricle ejection fraction, NYHA = New York Heart Association.

Nucleic acids research, Jan 24, 2015

Oligonucleotide aptamers represent a novel platform for creating ligands with desired specificity... more Oligonucleotide aptamers represent a novel platform for creating ligands with desired specificity, and they offer many potentially significant advantages over monoclonal antibodies in terms of feasibility, cost, and clinical applicability. However, the isolation of high-affinity aptamer ligands from random oligonucleotide pools has been challenging. Although high-throughput sequencing (HTS) promises to significantly facilitate systematic evolution of ligands by exponential enrichment (SELEX) analysis, the enormous datasets generated in the process pose new challenges for identifying those rare, high-affinity aptamers present in a given pool. We show that emulsion PCR preserves library diversity, preventing the loss of rare high-affinity aptamers that are difficult to amplify. We also demonstrate the importance of using reference targets to eliminate binding candidates with reduced specificity. Using a combination of bioinformatics and functional analyses, we show that the rate of am...

Hemangiomas and Vascular Malformations, 2015

The phlebolymphedema of the limbs is a disorder with frequent occurrence in its various forms, wh... more The phlebolymphedema of the limbs is a disorder with frequent occurrence in its various forms, which by its very nature tends to worsen and has a tendency to complications and clinical exacerbation.

Veins and Lymphatics, 2012

Hyperhomocysteinemia is a widely recognized, although not yet entirely understood, risk factor fo... more Hyperhomocysteinemia is a widely recognized, although not yet entirely understood, risk factor for cardiovascular disease. Particularly, the complex relationships between age, hyperhomocysteinemia, predisposing genetic factors and peripheral vascular diseases have not been fully evaluated. Our contribution to this issue is a retrospective analysis of a large series of patients with peripheral arterial, venous and lymphatic disease, and of their blood relatives, with special reference to homocysteine plasma levels, age and methylenetetrahydrofolate reductase (MTHFR) polymorphisms. Serum homocysteine was measured in 477 patients (286 males, 191 females, age range 19-78 years) with various vascular clinical conditions: postphlebitic syndrome (46) recurrent venous ulcers (78), arterial diseases (101) primary lymphoedema (87), secondary lymphoedema (161) and outlet thoracic syndrome (4), and in 50 normal controls. A MTHFR study for polymorphisms was carried on in the subjects with homocysteine values exceeding 15 mol/L. Serum homocysteine determination and MTHFR polymorphism studies were performed also in 1430 healthy blood related relatives (mainly siblings, descendents and sibling descendents) of the subjects with hyperhomocysteinemia and MTHFR polymorphisms. We found MTHFR polymorphisms in 20% of controls and in 69.3%, 69.5% and 53.8% of hyperhomocysteinemic subjects with arterial diseases, postphlebitic syndrome and venous ulcers, respectively. As expected, the percentage of hyperhomocysteinemia in patients with secondary lym

PLoS ONE, 2014

Recognition of microbial components via innate receptors including the C-type lectin receptor Dec... more Recognition of microbial components via innate receptors including the C-type lectin receptor Dectin-1, together with the inflammatory environment, programs dendritic cells (DCs) to orchestrate the magnitude and type of adaptive immune responses. The exposure to b-glucan, a known Dectin-1 agonist and component of fungi, yeasts, and certain immune support supplements, activates DCs to induce T helper (Th)17 cells that are essential against fungal pathogens and extracellular bacteria but may trigger inflammatory pathology or autoimmune diseases. However, the exact mechanisms of DC programming by b-glucan have not yet been fully elucidated. Using a gene expression/perturbation approach, we demonstrate that in human DCs b-glucan transcriptionally activates via an interleukin (IL)-1-and inflammasome-mediated positive feedback late-induced genes that bridge innate and adaptive immunity. We report that in addition to its known ability to directly prime T cells toward the Th17 lineage, IL-1 by promoting the transcriptional cofactor inhibitor of kB-f (IkB-f) also programs b-glucan-exposed