Papers by Maurizio Sorice
Cells
Background: Heparanase (HPSE) is an endo-β-glucuronidase that cleaves heparan sulfate side chains... more Background: Heparanase (HPSE) is an endo-β-glucuronidase that cleaves heparan sulfate side chains, leading to the disassembly of the extracellular matrix, facilitating cell invasion and metastasis dissemination. In this research, we investigated the role of a new HPSE inhibitor, RDS 3337, in the regulation of the autophagic process and the balance between apoptosis and autophagy in U87 glioblastoma cells. Methods: After treatment with RDS 3337, cell lysates were analyzed for autophagy and apoptosis-related proteins by Western blot. Results: We observed, firstly, that LC3II expression increased in U87 cells incubated with RDS 3337, together with a significant increase of p62/SQSTM1 levels, indicating that RDS 3337 could act through the inhibition of autophagic-lysosomal flux of LC3-II, thereby leading to accumulation of lipidated LC3-II form. Conversely, the suppression of autophagic flux could activate apoptosis mechanisms, as revealed by the activation of caspase 3, the increased l...
Erythrocytes, Dielectric Measurements, Flow-Cytometric M easurem ents Alterations in the electric... more Erythrocytes, Dielectric Measurements, Flow-Cytometric M easurem ents Alterations in the electrical passive param eters of red blood cell mem branes occurring during storage have been investigated by means of two different experimental approaches, i.e., ra diowave dielectric spectroscopy measurem ents and flow-cytometric measurements. We ob served a correlation between the appearance of phosphatidylserine molecules in the outer leaflet of the cell membrane and the occurrence of a change in the electrical passive mem brane parameters. The electrical reorganization of the membrane, resulting in an increase of its conductivity and permittivity after 5-7 days from blood storage, can be considered as a precursory event for the loss of asymmetry in the lipid distribution across red blood cell membrane.
![Research paper thumbnail of HMGB1 expression in leukocytes as a biomarker of cellular damage induced by [99mTc]Tc-HMPAO-labelling procedure: A quality control study](https://a.academia-assets.com/images/blank-paper.jpg)
Nuclear Medicine and Biology, May 1, 2021
PURPOSE Autologous White Blood Cells (WBC) scintigraphy is based on a multi-step sequence of cell... more PURPOSE Autologous White Blood Cells (WBC) scintigraphy is based on a multi-step sequence of cell separation and radiolabelling. Besides in vivo imaging quality control, no molecular tool is available to evaluate WBC damage secondary to cell manipulation. High Mobility Group Box 1 (HMGB1) is a protein of the alarmins family, secreted by innate immune cells and released from the nucleus of damaged cells following different types of injury. Aim of this study was to evaluate HMGB1 levels in WBC cytosolic extracts (CE) before and after [99mTc]Tc-HMPAO labelling procedure, as a biomarker of induced WBC damage. PROCEDURES Patients with suspect of prosthetic joint infection were prospectively enrolled. HMGB1 levels were evaluated by immunoblotting analysis in plasma (t0), and in WBC-CE before (t1) and after (t2) [99mTc]Tc-HMPAO labelling. Blood samples from healthy subjects were evaluated under the same procedure. RESULTS Twenty consecutive patients referred for WBC scintigraphy and ten controls were enrolled. HMGB1 levels were significantly upregulated both in plasma (t0) and in circulating WBC-CE (t1) from patients compared to controls (p < 0.0001). Otherwise, WBC-CE from [99mTc]Tc-HMPAO-labelled leukocyte concentrate (t2) did not show significant changes in HMGB1 levels compared to the cold leukocyte sample (t1). CONCLUSIONS The evaluation of HMGB1 levels in WBC-CE from each subject after radiolabelling with [99mTc]Tc-HMPAO did not show significant changes compared to the cold cellular sample. These results further prove the reliability of [99mTc]Tc-HMPAO leukocyte radiolabelling procedure in terms of cell viability and suggest that the monitoring of this alarmin may represent a specific tool to evaluate a secondary damage of WBC induced by radiolabelling procedure. In addition, significant upregulation of HMGB1 levels was found in WBC-CE and in plasma from patients with suspect of PJI - compared to healthy donors - reasonably related to their underlying inflammatory/infective condition.
Blood, Jun 9, 2011
In chronic disorders related to endothelial cell dysfunction, plasma  2 glycoprotein I ( 2 GPI)... more In chronic disorders related to endothelial cell dysfunction, plasma  2 glycoprotein I ( 2 GPI) plays a role as a target antigen of pathogenetic autoimmune responses. However, information is still lacking to clarify why  2 GPI triggers autoimmunity. It is possible that posttranslational modification of the protein, such as nonenzymatic glycosylation, leads to the formation of advanced glycation end products (AGEs). The aim of our study was to explore whether glucose-modified  2 GPI is able to interact and activate monocytederived immature dendritic cells (iDCs) from healthy human donors. SDS-PAGE and spectrofluorometric analyses indicated that  2 GPI incubated with glucose was sugar modified, and that this modification likely consisted of AGE formation, resulting in AGE- 2 GPI. AGE- 2 GPI caused phenotypical and functional maturation of iDCs involving the activation of p38 MAPK, ERK, and NF-B. It also induced on DCs a significant up-regulation of RAGE, the receptor for AGEs. Evidence for RAGE involvement comes from blocking experiments with an anti-RAGE mAb, confocal analysis, and coimmunoprecipitation experiments. AGE- 2 GPI-stimulated DCs had increased allostimulatory ability and primed naive T lymphocytes toward a Th2 polarization. These findings might explain in part the interactive role of  2 GPI, AGEs, and DCs in chronic disorders related to endothelial cell dysfunction. (Blood. 2011;117(23):6152-6161)
Biomedicines, Dec 2, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Journal of Alzheimer's Disease, Jan 18, 2022
Specific protein misfolding and aggregation are mechanisms underlying various neurodegenerative d... more Specific protein misfolding and aggregation are mechanisms underlying various neurodegenerative diseases such as prion disease and Alzheimer's disease (AD). The misfolded proteins are involved in prions, amyloid- (A), tau, and ␣synuclein disorders; they share common structural, biological, and biochemical characteristics, as well as similar mechanisms of aggregation and self-propagation. Pathological features of AD include the appearance of plaques consisting of deposition of protein A and neurofibrillary tangles formed by the hyperphosphorylated tau protein. Although it is not clear how protein aggregation leads to AD, we are learning that the cellular prion protein (PrP C) plays an important role in the pathogenesis of AD. Herein, we first examined the pathogenesis of prion and AD with a focus on the contribution of PrP C to the development of AD. We analyzed the mechanisms that lead to the formation of a high affinity bond between A oligomers (AOs) and PrP C. Also, we studied the role of PrP C as an AO receptor that initiates an AO-induced signal cascade involving mGluR5, Fyn, Pyk2, and eEF2K linking A and tau pathologies, resulting in the death of neurons in the central nervous system. Finally, we have described how the PrP C-AOs interaction can be used as a new potential therapeutic target for the treatment of PrP C-dependent AD.
Biomedicines, May 3, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Neuroscience, 2014
The cellular form of prion protein (PrPc) is a highly conserved cell surface GPI-anchored glycopr... more The cellular form of prion protein (PrPc) is a highly conserved cell surface GPI-anchored glycoprotein that was identified in cholesterol-enriched, detergent-resistant microdomains (‘rafts’) in neural and non-neural cells. Several physiological functions have been described for PrPC, including oxidative stress defense, metal ion homeostasis in the brain, and neuroprotection. Aβ oligomers, but not monomers or fibrils, bind tightly to PrPc (Kd ~ 0.4 nM) and, in hippocampal slices, PrPc PrPc is required for Aβ oligomer-mediated inhibition of long-term potentiation. Our results suggest that the activity of PrPc-Aβ requires lipid rafts and the transmembrane receptor, low-density lipoprotein receptor-related protein (LRP1). LRP1 functions as an endocytic receptor for a broad range of structurally and functionally diverse ligands. LRP1 also functions in cell sig¬naling, directly, in response to ligand-binding, and indirectly, by regulating levels of other signaling receptors. In neurons and neurite-generating cell lines, ligands control the sig¬naling activity of LRP1 by directing the co-receptors that are recruited into a functional signaling complex with LRP1. LRP1 is also known to control surface and biosynthetic trafficking of PrPc in neurons. Our results show that PrPc is strictly associated with gangliosides in lipid rafts in neuroblastoma cells. Scanning confocal microscopy analysis revealed co-localization of PrPc with GM1, as well as TrkA with GM1, indicating the existence of a glycosphingolipid-enriched molecular complex. In order to analyze the mechanism by which recombinant PrPc initiates cell signaling in SK-N-BE2 and PC12 neuron-like cells, we examined ERK1/2 phosphorylation. ERK1/2 was robustly phosphorylated in cells that were treated with recombinant PrPc. ERK1/2 activation required LRP1 and was strictly dependent on the integrity of rafts. ERK1/2 activation was blocked by altering sphingolipid metabolism with fumonisin B1 or by disruption of lipid microdomains with methyl-β-cyclodextrin. These findings support a model in which LRP1 initiates signal transduction specifically in lipid rafts, where it forms multimolecular complexes that may include PrPc, TrkA and gangliosides. Although association of LRP1 with lipid rafts has been reported before, this is the first study to demonstrate a potentially significant LRP1 activity that depends on its distribution between rafts, clathrin-coated pits and other membrane domains
Scandinavian Journal of Immunology, 1994
In this study we analysed the relationship between anti‐lymphocytic ganglioside antibodies and an... more In this study we analysed the relationship between anti‐lymphocytic ganglioside antibodies and anti‐lymphocyte antibodies in AIDS patients. Anti‐lymphocytic ganglioside antibodies were detected by thin layer chromatography (TLC) immunostaining; three colour flow cytometry was used to analyse circulating antibodies against different lymphocyte subsets.Anti‐lymphocytic ganglioside antibodies were detected in 23 out of 49 AIDS patient sera (46.9%). All positive sera reacted selectively with the GM3 comigrating band from AIDS lymphocytes. Twenty two out of the 23 anti‐lymphocytic GM3 positive sera also had antibodies against CD4+ T cells, versus 17/26 anti‐GM3 negative. Furthermore, patients with lymphocytic GM3 antibodies showed a significantly higher antibody reactivity against CD4+ T cells than patients in which these antibodies were not detected. The absorption tests revealed that preincubation of positive sera with GM3 was followed by a decrease in the reaction with target lymphocy...
Infection, 1995
In this study the presence of brain antiganglioside antibodies in the cerebrospinal fluid (CSF) o... more In this study the presence of brain antiganglioside antibodies in the cerebrospinal fluid (CSF) of patients with HIV infection was analysed. CSF samples were collected from 45 patients with AIDS and from 45 anti-HIV negative subjects, 15 of whom presented aseptic meningitis. Nineteen AIDS patients had clinically well-documented encephalopathy. Thirteen of these patients had white matter lesions shown by magnetic resonance imaging (MRI). Both IgG and IgM antiganglioside antibodies were detected by immunostaining on thin layer chromatography plates in three CSF samples from AIDS patients with progressive encephalopathy with signs of a diffuse demyelination, as revealed by MRI. Two of these CSF samples reacted specifically with GM3, GM1 and GD1a and one with GD1a. In none of the HIV infected patients without demyelinating encephalopathy, but with opportunistic infections or cerebral lymphoma, nor in the anti-HIV negative control subjects were antiganglioside antibodies detected. No association with JCV DNA, CMV DNA, EBV DNA, detected by nested PCR, nor HIV antigen p24 was found. These findings show the presence of brain antiganglioside antibodies in the CSF of AIDS patients for the first time. However, the findings do not suggest relating the presence of these antibodies to HIV encephalopathy or particular viral agents, but indicate that the antibodies are detectable in subjects with progressive encephalopathy with a diffuse demyelination.
Clinical and Experimental Immunology, 2021
Current literature regarding systemic autoimmune diseases in X-chromosome aneuploidies is scarce ... more Current literature regarding systemic autoimmune diseases in X-chromosome aneuploidies is scarce and limited to case reports. Our aim was to evaluate the frequency of anti-nuclear (ANAs), extractable nuclear (ENA), anti-double-stranded DNA (dsDNAs), anti-smooth muscle (ASMAs) and anti-mitochondrial (AMAs) antibodies in a large cohort of adults with Klinefelter's syndrome (KS, 47,XXY) and rare higher-grade sex chromosome aneuploidies (HGAs) for the first time. Sera from 138 X-chromosome aneuploid patients [124 adult patients with 47,XXY KS and 14 patients with HGA (six children, eight adults)] and 50 age-matched 46,XY controls were recruited from the Sapienza University of Rome (2007–17) and tested for ANAs, ENAs, anti-dsDNAs, ASMAs and AMAs. Non-organ-specific immunoreactivity was found to be significantly higher in patients with 47,XXY KS (14%) than in the controls (2%, p = 0.002). Among all the antibodies investigated, only ANAs were observed significantly more frequently in p...
Central nervous system (CNS) involvement in systemic lupus erythematosus (SLE) is reported in abo... more Central nervous system (CNS) involvement in systemic lupus erythematosus (SLE) is reported in about 50% of patients. Among the neuropsychiatric features of SLE, myelopathy, including acute transverse myelitis (ATM) or acute longitudinal myelitis (ALM), represents an uncommon event. A possible vascular aetiology of SLE myelopathies has been hypothesized and it seems to be much more associated to SLE-associated antiphospholipid syndrome (APS). Furthermore, a possible infectious cause of ATM or ALM in healthy subjects has been described. SLE patients are susceptible to infection due to the disease itself or to the immunosuppressive therapy. Cryptococci non-neoformans have been rarely associated to infections in humans. Here we describe the case of a 47-year-old woman with SLE and Sjögren Syndrome who developed an ALM concurrently with a Cryptococcus laurentii pneumonia. The patient was treated with antimycotics, high doses of glucocorticoids and intravenous immunoglobulins with a significant clinical and radiological improvement. As far as we know, this is the first case of Cryptococcus laurentii infection and ALM in a patient with SLE who later developed a seronegative APS. Even though myelopathy may be considered primarily associated to SLE, a possible role of the infection in ALM development cannot be excluded.
Biomedicines
Antiphospholipid antibody syndrome is an autoimmune disease characterized by thrombosis and/or pr... more Antiphospholipid antibody syndrome is an autoimmune disease characterized by thrombosis and/or pregnancy morbidity in association with circulating antiphospholipid antibodies, mainly anti-β2 glycoprotein 1 antibodies (anti-β2-GPI antibodies). Previous studies demonstrated that the signaling pathway may involve lipid rafts, plasma membrane microdomains enriched in glycosphingolipid and cholesterol. In this study, we analyzed the signaling pathway of LRP8/ApoER2, a putative receptor of anti-β2-GPI antibodies, through lipid rafts in human endothelial cells. LRP8, Dab2 and endothelial nitric oxide synthase (e-NOS) phosphorylation were evaluated using Western blot, Nitric Oxide (NO) production with cytofluorimetric analysis, LRP8 enrichment in lipid rafts via sucrose gradient fractionation, and scanning confocal microscopy analysis of its association with ganglioside GM1 was also conducted. The analyses demonstrated that affinity-purified anti-β2-GPI antibodies induced LRP8 and Dab-2 pho...
Journal of Thrombosis and Haemostasis, 2022
ORCID Nigel Mackman https://orcid.org/0000-0002-9170-7700 Yohei Hisada https://orcid.org/0000-000... more ORCID Nigel Mackman https://orcid.org/0000-0002-9170-7700 Yohei Hisada https://orcid.org/0000-0001-9157-0524 R E FE R E N C E S 1. Capozzi A, Riitano G, Recalchi S, et al. Effect of heparanase inhibitor on tissue factor overexpression in platelets and endothelial cells induced by antibeta2GPI antibodies. J Thromb Haemost. 2021;19:23022313. 2. Rosell A, Moser B, Hisada Y, et al. Evaluation of different commercial antibodies for their ability to detect human and mouse tissue factor by western blotting. Res Pract Thromb Haemost. 2020;4:10131023. 3. Capozzi A, Manganelli V, Riitano G, et al. Tissue factor overexpression in platelets of patients with antiphospholipid syndrome: induction role of antibeta2GPI antibodies. Clin Exp Immunol. 2019;196:5966. 4. Mackman N, Luther T. Platelet tissue factor: to be or not to be. Thromb Res. 2013;132:35. 5. Osterud B, Bouchard BA. Detection of tissue factor in platelets: why is it so troublesome? Platelets. 2019;30:15. 6. Brambilla M, Facchinetti L, Canzano P, et al. Human megakaryocytes confer tissue factor to a subset of shed platelets to stimulate thrombin generation. Thromb Haemost. 2015;114:579592. 7. Daugherty A, Hegele RA, Mackman N, Rader DJ, Schmidt AM, Weber C. Complying with the National Institutes of Health guidelines and principles for rigor and reproducibility: refutations. Arterioscler Thromb Vasc Biol. 2016;36:13031304. 8. Uhlen M, Bandrowski A, Carr S, et al. A proposal for validation of antibodies. Nat Methods. 2016;13:823827. 9. Kobayashi S, Koizume S, Takahashi T, et al. Tissue factor and its procoagulant activity on cancerassociated thromboembolism in pancreatic cancer. Cancer Sci. 2021. https://doi.org/10.1111/ cas.15106. Online ahead of print. 10. Morrissey JH, Fakhrai H, Edgington TS. Molecular cloning of the cDNA for tissue factor, the cellular receptor for the initiation of the coagulation protease cascade. Cell. 1987;50:129135.
Parasitology, Aug 1, 1992
This study was undertaken to assess whether glycolipid antigens (particularly gangliosides) are a... more This study was undertaken to assess whether glycolipid antigens (particularly gangliosides) are associated withPneumocystis cariniiobtained from human lungs. Gangliosides were extracted, purified in high performance thin-layer chromatography and stained with resorcinol. Two resorcinol-positive bands, co-migrating with GM1and GD1awere demonstrated, suggesting the existence of ganglioside molecules onP. carinii. No resorcinol-positive bands were revealed in the pulmonary control tissue. In addition, an antiserum obtained from rabbits immunized withP. cariniiantigen reacted with gangliosides GM1, and GD1a, as revealed by a dot immunobinding assay. This reactivity was inhibited by first incubating the antiserum with ganglioside micelles. Furthermore, anti-glycosphingolipid antibodies (aGM1) reacted with the bands of 200 and 55 kDa ofP. cariniiantigen. These results suggest that ganglioside antigens expressed onP. cariniican trigger specific immune responses.
Clinical and Experimental Immunology, Feb 7, 2005
Lyso(bis)phosphatidic acid (LBPA) is a novel antigenic target in antiphospholipid syndrome (APS) ... more Lyso(bis)phosphatidic acid (LBPA) is a novel antigenic target in antiphospholipid syndrome (APS) and antibodies directed against LBPA (aLBPA) have been detected in sera from APS patients. In this study we first evaluated aLBPA in comparison with the most widely used methods (i.e. anticardiolipin [(aCL)-enzyme-linked immunosorbent assay (ELISA)] and antibeta-2glycoprotein-I antibodies (ab b b b 2-GPI-ELISA) utilized to detect antiphospholipid antibodies in patients with primary or secondary APS, systemic lupus erythematosus, chronic HCV infection and healthy subjects. We then assessed the relationship between aLBPA, lupus anticoagulant (LAC) and the main clinical manifestations of APS. Finally, we evaluated the presence of 'pure' (i.e. b b b b 2-GPI-independent) aLBPA in patients with APS and controls. The results indicate that aLBPA as well as ab b b b 2-GPI display higher specificity but lower sensitivity for APS compared to aCL. Moreover, serum aLBPA correlate closely with aCL and ab b b b 2-GPI in APS patients and are strictly associated with LAC positivity. We demonstrate that b b b b 2-GPI binds to LBPA with affinity similar to CL, and antibodies able to react with phosholipid-protein complex exist; however, 'pure' aLBPA can also be detected in sera of APS patients. Altogether these data confirm that LBPA may be an antigenic target in APS and that aLBPA are serological markers of APS with similar sensitivity and specificity compared to ab b b b 2-GPI. However, the clinical utility of aLBPA detection alone or in combination with aCL and/or ab b b b 2-GPI remains to be elucidated in larger and longitudinal studies.
Prion, Sep 1, 2012
T he cellular form of prion protein (PrP C) is a highly conserved cell surface GPI-anchored glyco... more T he cellular form of prion protein (PrP C) is a highly conserved cell surface GPI-anchored glycoprotein that was identified in cholesterol-enriched, detergent-resistant microdomains named "rafts." The association with these specialized portions of the cell plasma membrane is required for conversion of PrP C to the transmissible spongiform encephalopathy-associated protease-resistant isoform. Usually, PrP C is reported to be a plasma membrane protein, however several studies have revealed PrP C as an interacting protein mainly with the membrane/organelles, as well as with cytoskeleton network. Recent lines of evidence indicated its association with ER lipid raft-like microdomains for a correct folding of PrP C , as well as for the export of the protein to the Golgi and proper glycosylation. During cell apoptosis, PrP C can undergo intracellular re-localization, via ER-mitochondria associated membranes (MAM) and microtubular network, to mitochondrial raft-like microdomains, where it induced the loss of mitochondrial membrane potential and cytochrome c release, after a contained raise of calcium concentration. We suggest that PrP C may play a role in the multimolecular signaling complex associated with cell apoptosis. Lipid rafts and their components may, thus, be investigated as pharmacological targets of interest, introducing a novel and innovative task in modern pharmacology, i.e., the development of glycosphingolipid targeted drugs.
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Papers by Maurizio Sorice