46 results on '"Sgarbanti M"'
Search Results
2. I kappa B kinase (IKK)-epsilon targets interferon regulatory factor 1 in activated T lymphocytes
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Sgarbanti, M, Marsili, G, Remoli, Al, Stellacci, E, Mai, Antonello, Rotili, Dante, Perrotti, E, Acchioni, C, Orsatti, R, Iraci, N, Ferrari, M, Borsetti, A, Hiscott, J, and Battistini, A.
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- 2014
3. Epigenetic drug Gar1041 in combination with antiretroviral therapy transiently reduces the proviral DNA reservoir in SIVmac251‐infected macaques
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Lewis, Mg, Norelli, S, Chomont, N, De Fonseca, S, Sgarbanti, M, Collins, M, Chirullo, B, Yalley?Ogunro, J, Greenhouse, J, Palamara, At, Garaci, E, and Savarino, A
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DNA -- Health aspects -- Physiological aspects ,Leukemia -- Drug therapy -- Genetic aspects ,Antineoplastic agents -- Testing -- Dosage and administration ,Antiviral agents -- Testing -- Dosage and administration ,Epigenetic inheritance -- Health aspects -- Physiological aspects ,Antimitotic agents -- Testing -- Dosage and administration ,Health - Abstract
16‐17 July 2010, International AIDS Society’s Workshop “Towards a Cure”: HIV Reservoirs and Strategies to Control Them, Vienna, Austria, Background It was recently hypothesized that the lentiviral reservoir in central memory (T[sub.CM]) and transitional memory (T[sub.TM]) CD4+ cells could be restricted by new therapies targeting pathways downstream of homeostatic [...]
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- 2010
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4. IRF-7: New Role in the Regulation of Genes Involved in Adaptive Immunity
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SGARBANTI, M., primary, MARSILI, G., additional, REMOLI, A. L., additional, ORSATTI, R., additional, and BATTISTINI, A., additional
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- 2007
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5. Review: IRF Regulation of HIV-1 Long Terminal Repeat Activity
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Battistini, A., primary, Marsili, G., additional, Sgarbanti, M., additional, Ensoli, B., additional, and Hiscott, J., additional
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- 2002
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6. Co-Infections and Superinfections between HIV-1 and Other Human Viruses at the Cellular Level.
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Acchioni C, Sandini S, Acchioni M, and Sgarbanti M
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Co-infection or superinfection of the host by two or more virus species is a common event, potentially leading to viral interference, viral synergy, or neutral interaction. The simultaneous presence of two or more viruses, even distantly related, within the same cell depends upon viral tropism, i.e., the entry of viruses via receptors present on the same cell type. Subsequently, productive infection depends on the ability of these viruses to replicate efficiently in the same cellular environment. HIV-1 initially targets CCR5-expressing tissue memory CD4
+ T cells, and in the absence of early cART initiation, a co-receptor switch may occur, leading to the infection of naïve and memory CXCR4-expressing CD4+ T cells. HIV-1 infection of macrophages at the G1 stage of their cell cycle also occurs in vivo, broadening the possible occurrence of co-infections between HIV-1 and other viruses at the cellular level. Moreover, HIV-1-infected DCs can transfer the virus to CD4+ T cells via trans -infection. This review focuses on the description of reported co-infections within the same cell between HIV-1 and other human pathogenic, non-pathogenic, or low-pathogenic viruses, including HIV-2, HTLV, HSV, HHV-6/-7, GBV-C, Dengue, and Ebola viruses, also discussing the possible reciprocal interactions in terms of virus replication and virus pseudotyping.- Published
- 2024
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7. Cognitive assessment to optimize prediction of functional outcome in subacute hip fracture: a short-term prospective study.
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DI Monaco M, Sgarbanti M, Trombetta S, Gullone L, Bonardo A, Gindri P, Castiglioni C, Bardesono F, Milano E, and Massazza G
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- Humans, Prospective Studies, Activities of Daily Living, Prognosis, Cognition, Hip Fractures rehabilitation, Cognitive Dysfunction diagnosis, Cognitive Dysfunction etiology
- Abstract
Background: Cognitive impairment is a long-known negative prognostic factor after hip fracture. Cognition is usually screened by a single easy-to-administer bedside tool, but recent studies have shown that screening tests may be not enough to rule out cognitive impairment with an unfavorable prognostic role. Unfortunately, data on outcome prediction by further cognitive assessments is sparse., Aim: We focused on patients with subacute hip fracture defined cognitively intact or mildly impaired on the screening evaluation performed by the Short Portable Mental Status Questionnaire (SPMSQ). We hypothesized that each of 3 further cognitive tests could independently predict activities of daily living, with optimal prediction of function obtained by performing all three the tests., Design: Short-term prospective study., Setting: Rehabilitation ward., Population: Inpatients with subacute hip-fracture., Methods: Three cognitive tests were performed on admission to rehabilitation in the patients who made ≤4 errors on the SPMSQ: Montreal Cognitive Assessment (MoCA), Rey Auditory Verbal Learning Test (RAVLT, immediate and delayed recall) and Frontal Assessment Battery (FAB). We assessed activities of daily living by the Barthel index. Successful rehabilitation was defined with a Barthel Index Score ≥85., Results: Each of the three cognitive tests assessed before rehabilitation significantly predicted the Barthel index scores measured at the end of the rehabilitation course in our sample of 280 inpatients. However, only the MoCA score retained its significant predictive role when the scores from the three tests were included together as independent variables in a multiple regression model, with adjustments for a panel of potential confounders (P=0.007). The adjusted odds ratio to achieve successful rehabilitation for a seven-point change in MoCA score was 1.98 (CI 95% from 1.02 to 3.83; P=0.042)., Conclusions: Contrary to our hypothesis, MoCA but not RAVLT and FAB retained the prognostic role when the scores from the three tests were evaluated together as potential predictors of functional ability in activities of daily living., Clinical Rehabilitation Impact: In the presence of a normal (or mildly altered) score on the SPMSQ in subacute hip fracture, MoCA scores improve prediction of activities of daily living and should be routinely performed.
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- 2024
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8. Identification of Anti-Influenza A Compounds Inhibiting the Viral Non-Structural Protein 1 (NS1) Using a Type I Interferon-Driven Screening Strategy.
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Marsili G, Acchioni C, Remoli AL, Amatore D, Sgarbanti R, De Angelis M, Orsatti R, Acchioni M, Astolfi A, Iraci N, Puzelli S, Facchini M, Perrotti E, Cecchetti V, Sabatini S, Superti F, Agamennone M, Barreca ML, Hiscott J, Nencioni L, and Sgarbanti M
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- Humans, Viral Nonstructural Proteins metabolism, Antiviral Agents pharmacology, Antiviral Agents metabolism, Virus Replication, Influenza A Virus, H1N1 Subtype genetics, Interferon Type I metabolism, Influenza, Human drug therapy, Influenza A virus genetics
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There is an urgent need to identify efficient antiviral compounds to combat existing and emerging RNA virus infections, particularly those related to seasonal and pandemic influenza outbreaks. While inhibitors of the influenza viral integral membrane proton channel protein (M2), neuraminidase (NA), and cap-dependent endonuclease are available, circulating influenza viruses acquire resistance over time. Thus, the need for the development of additional anti-influenza drugs with novel mechanisms of action exists. In the present study, a cell-based screening assay and a small molecule library were used to screen for activities that antagonized influenza A non-structural protein 1 (NS1), a highly conserved, multifunctional accessory protein that inhibits the type I interferon response against influenza. Two potential anti-influenza agents, compounds 157 and 164 , were identified with anti-NS1 activity, resulting in the reduction of A/PR/8/34(H1N1) influenza A virus replication and the restoration of IFN-β expression in human lung epithelial A549 cells. A 3D pharmacophore modeling study of the active compounds provided a glimpse of the structural motifs that may contribute to anti-influenza virus activity. This screening approach is amenable to a broader analysis of small molecule compounds to inhibit other viral targets.
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- 2023
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9. A cellular screening platform, stably expressing DENV2 NS5, defines a novel anti-DENV mechanism of action of Apigenin based on STAT2 activation.
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Acchioni C, Acchioni M, Mancini F, Amendola A, Marsili G, Tirelli V, Gwee CP, Chan KW, Sandini S, Bisbocci M, Mysara M, ElHefnawi M, Sanchez M, Venturi G, Barreca ML, Manfroni G, Bresciani A, Vasudevan SG, and Sgarbanti M
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- Apigenin pharmacology, Luteolin pharmacology, Signal Transduction, STAT2 Transcription Factor genetics, STAT2 Transcription Factor metabolism, Viral Nonstructural Proteins genetics, Viral Nonstructural Proteins metabolism, Humans, Dengue Virus physiology
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Type I interferon (IFN-I) evasion by Dengue virus (DENV) is key in DENV pathogenesis. The non-structural protein 5 (NS5) antagonizes IFN-I response through the degradation of the signal transducer and activator of transcription 2 (STAT2). We developed a K562 cell-based platform, for high throughput screening of compounds potentially counteracting the NS5-mediated antagonism of IFN-I signaling. Upon a screening with a library of 1220 approved drugs, 3 compounds previously linked to DENV inhibition (Apigenin, Chrysin, and Luteolin) were identified. Luteolin and Apigenin determined a significant inhibition of DENV2 replication in Huh7 cells and the restoration of STAT2 phosphorylation in both cell systems. Apigenin and Luteolin were able to stimulate STAT2 even in the absence of infection. Despite the "promiscuous" and "pan-assay-interfering" nature of Luteolin, Apigenin promotes STAT2 Tyr 689 phosphorylation and activation, highlighting the importance of screening for compounds able to interact with host factors, to counteract viral proteins capable of dampening innate immune responses., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2023
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10. New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis.
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Mariotti S, Chiantore MV, Teloni R, Iacobino A, Capocefalo A, Michelini Z, Borghi M, Baggieri M, Marchi A, Bucci P, Gioacchini S, D'Amelio R, Brouwer PJM, Sandini S, Acchioni C, Sgarbanti M, Di Virgilio A, Grasso F, Cara A, Negri D, Magurano F, Di Bonito P, and Nisini R
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The emergence of the new pathogen SARS-CoV-2 determined a rapid need for monoclonal antibodies (mAbs) to detect the virus in biological fluids as a rapid tool to identify infected individuals to be treated or quarantined. The majority of commercially available antigenic tests for SARS-CoV-2 rely on the detection of N antigen in biologic fluid using anti-N antibodies, and their capacity to specifically identify subjects infected by SARS-CoV-2 is questionable due to several structural analogies among the N proteins of different coronaviruses. In order to produce new specific antibodies, BALB/c mice were immunized three times at 20-day intervals with a recombinant spike (S) protein. The procedure used was highly efficient, and 40 different specific mAbs were isolated, purified and characterized, with 13 ultimately being selected for their specificity and lack of cross reactivity with other human coronaviruses. The specific epitopes recognized by the selected mAbs were identified through a peptide library and/or by recombinant fragments of the S protein. In particular, the selected mAbs recognized different linear epitopes along the S1, excluding the receptor binding domain, and along the S2 subunits of the S protein of SARS-CoV-2 and its major variants of concern. We identified combinations of anti-S mAbs suitable for use in ELISA or rapid diagnostic tests, with the highest sensitivity and specificity coming from proof-of-concept tests using recombinant antigens, SARS-CoV-2 or biological fluids from infected individuals, that represent important additional tools for the diagnosis of COVID-19.
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- 2023
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11. Kidney transplant in patients with atypical hemolytic uremic syndrome in the anti-C5 era: single-center experience with tailored Eculizumab.
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Ardissino G, Cresseri D, Tel F, Giussani A, Salardi S, Sgarbanti M, Strumbo B, Testa S, Capone V, Griffini S, Grovetti E, Cugno M, Belingheri M, Tamburello C, Rodrigues EM, Perrone M, Cardillo M, Corti G, Consonni D, Furian L, Tedeschi S, Messa P, and Beretta C
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- Antibodies, Monoclonal, Humanized therapeutic use, Female, Humans, Plasmapheresis, Atypical Hemolytic Uremic Syndrome diagnosis, Atypical Hemolytic Uremic Syndrome drug therapy, Kidney Transplantation adverse effects
- Abstract
Rationale and Objective: Patients with atypical hemolytic uremic syndrome (aHUS) have long been considered ineligible for kidney transplantation (KTx) in several centers due to the high risk of disease recurrence, graft loss and life-threatening complications. The availability of Eculizumab (ECU) has now overcome this problem. However, the best approach towards timing, maintenance schedule, the possibility of discontinuation and patient monitoring has not yet been clearly established., Study Design: This is a single center case series presenting our experience with KTx in aHUS., Setting and Participants: This study included 26 patients (16 females) with a diagnosis of aHUS, who spent a median of 5.5 years on kidney replacement therapy before undergoing KTx. We compared the aHUS relapse rate in three groups of patients who underwent KTx: patients who received no prophylaxis, patients who underwent plasma exchange, those who received Eculizumab prophylaxis. Complement factor H-related disease was by far the most frequent etiology (n = 19 patients)., Results: Untreated patients and patients undergoing pre-KTx plasma exchange prophylaxis had a relapse rate of 0.81 (CI 0.30-1.76) and 3.1 (CI 0.64-9.16) events per 10 years cumulative observation, respectively, as opposed to 0 events among patients receiving Eculizumab prophylaxis. The time between Eculizumab doses was tailored based on classic complement pathway activity (target to < 30%). Using this strategy, 12 patients are currently receiving Eculizumab every 28 days, 5 every 24-25 days, and 3 every 21 days., Conclusion: Our experience supports the prophylactic use of Eculizumab in patients with a previous history of aHUS undergoing KTx, especially when complement dysregulation is well documented by molecular biology., (© 2021. Italian Society of Nephrology.)
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- 2021
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12. Fighting HIV-1 Persistence: At the Crossroads of "Shoc-K and B-Lock".
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Acchioni C, Palermo E, Sandini S, Acchioni M, Hiscott J, and Sgarbanti M
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Despite the success of highly active antiretroviral therapy (HAART), integrated HIV-1 proviral DNA cannot be eradicated from an infected individual. HAART is not able to eliminate latently infected cells that remain invisible to the immune system. Viral sanctuaries in specific tissues and immune-privileged sites may cause residual viral replication that contributes to HIV-1 persistence. The "Shock or Kick, and Kill" approach uses latency reversing agents (LRAs) in the presence of HAART, followed by cell-killing due to viral cytopathic effects and immune-mediated clearance. Different LRAs may be required for the in vivo reactivation of HIV-1 in different CD4
+ T cell reservoirs, leading to the activation of cellular transcription factors acting on the integrated proviral HIV-1 LTR. An important requirement for LRA drugs is the reactivation of viral transcription and replication without causing a generalized immune activation. Toll-like receptors, RIG-I like receptors, and STING agonists have emerged recently as a new class of LRAs that augment selective apoptosis in reactivated T lymphocytes. The challenge is to extend in vitro observations to HIV-1 positive patients. Further studies are also needed to overcome the mechanisms that protect latently infected cells from reactivation and/or elimination by the immune system. The Block and Lock alternative strategy aims at using latency promoting/inducing agents (LPAs/LIAs) to block the ability of latent proviruses to reactivate transcription in order to achieve a long term lock down of potential residual virus replication. The Shock and Kill and the Block and Lock approaches may not be only alternative to each other, but, if combined together (one after the other), or given all at once [namely "Shoc-K(kill) and B(block)-Lock"], they may represent a better approach to a functional cure.- Published
- 2021
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13. Short- and Long-Term Immunological Responses in Chronic HCV/HIV Co-Infected Compared to HCV Mono-Infected Patients after DAA Therapy.
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Farcomeni S, Moretti S, Fimiani C, Sulekova LF, Vescio F, Sernicola L, Maggiorella MT, Remoli AL, Picconi O, Mosca L, Esvan R, Biliotti E, Ciccozzi M, Sgarbanti M, Taliani G, and Borsetti A
- Abstract
Background: Direct-acting antivirals (DAAs) treatment, although highly efficacious for the treatment of hepatitis C virus (HCV) infection, may not completely reconstitute the HCV-mediated dysregulated immune system, especially in patients co-infected with human immunodeficiency virus (HIV) and HCV., Objectives: We aimed to evaluate the impact of HCV eradication following DAA therapy on the immune system and liver disease improvement through comparative monitoring of 10 HCV mono-infected and 10 HCV/HIV co-infected patients under combined antiretroviral therapy (cART). Early and late longitudinal phenotypic changes in peripheral blood mononuclear cell (PBMC) subsets, T-cell activation, differentiation and exhaustion, as well as inflammatory biomarkers, indoleamine 2-3 dioxygenase (IDO) activity, and liver stiffness, APRI and FIB-4 scores were assessed., Materials and Methods: Samples were obtained at baseline (T0), week 1 (T1), week 2 (T2), week 12 (T3, end of treatment, EOT), and month 9 (T4, end of follow-up, 36 weeks post EOT)., Results: All patients achieved a sustained virological response (SVR 12) after DAA treatment. Overall, changes of the T-cell immune phenotypes were greater in HCV/HIV co-infected than in HCV mono-infected, due to an increase in CD4+ and CD8+ T-cell percentages and of CD8+ T-cell activation and memory markers, in particular at the end of follow-up. On the other end, HCV mono-infected showed changes in the activation profile and in the memory CD4+ T-cell compartment. In HCV/HIV co-infected, a decrease in the IDO activity by DAA treatment was observed; conversely, in HCV mono-infected, it resulted unmodified. Regarding inflammatory mediators, viral suppression was associated with a reduction in IP-10 levels, while interferon regulatory factor (IRF)-7, interferon (IFN)-β, and interferon (IFN)-γ levels were downregulated during therapy and increased post therapy. A decrease in liver stiffness, APRI, and FIB-4 scores was also observed., Conclusions: Our study suggests that, although patients achieved HCV eradication, the immune activation state in both HCV mono-infected and HCV/HIV co-infected patients remains elevated for a long time after the end of DAA therapy, despite an improvement of liver-specific outcomes, meanwhile highlighting the distinct immunophenotypic and inflammatory biomarker profile between the groups of patients.
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- 2021
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14. CRISPR/Cas9 Ablation of Integrated HIV-1 Accumulates Proviral DNA Circles with Reformed Long Terminal Repeats.
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Lai M, Maori E, Quaranta P, Matteoli G, Maggi F, Sgarbanti M, Crucitta S, Pacini S, Turriziani O, Antonelli G, Heeney JL, Freer G, and Pistello M
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- CRISPR-Associated Protein 9, Gene Editing, Gene Expression Regulation, Viral, Genetic Therapy, HEK293 Cells, HIV Infections virology, Humans, RNA, Guide, CRISPR-Cas Systems genetics, CRISPR-Cas Systems, DNA, Circular, HIV-1 genetics, Proviruses genetics, Terminal Repeat Sequences
- Abstract
Gene editing may be used to excise the human immunodeficiency virus type 1 (HIV-1) provirus from the host cell genome, possibly eradicating the infection. Here, using cells acutely or latently infected by HIV-1 and treated with long terminal repeat (LTR)-targeting CRISPR/Cas9, we show that the excised HIV-1 provirus persists for a few weeks and may rearrange in circular molecules. Although circular proviral DNA is naturally formed during HIV-1 replication, we observed that gene editing might increase proviral DNA circles with restored LTRs. These extrachromosomal elements were recovered and probed for residual activity through their transfection in uninfected cells. We discovered that they can be transcriptionally active in the presence of Tat and Rev. Although confirming that gene editing is a powerful tool to eradicate HIV-1 infection, this work highlights that, to achieve this goal, the LTRs must be cleaved in several pieces to avoid residual activity and minimize the risk of reintegration in the context of genomic instability, possibly caused by the off-target activity of Cas9. IMPORTANCE The excision of HIV-1 provirus from the host cell genome has proven feasible in vitro and, to some extent, in vivo . Among the different approaches, CRISPR/Cas9 is the most promising tool for gene editing. The present study underlines the remarkable effectiveness of CRISPR/Cas9 in removing the HIV-1 provirus from infected cells and investigates the fate of the excised HIV-1 genome. This study demonstrates that the free provirus may persist in the cell after editing and in appropriate circumstances may reactivate. As an episome, it might be transcriptionally active, especially in the presence of Tat and Rev. The persistence of the HIV-1 episome was strongly decreased by gene editing with multiple targets. Although gene editing has the potential to eradicate HIV-1 infection, this work highlights a potential issue that warrants further investigation.
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- 2021
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15. Isolation and Characterization of Mouse Monoclonal Antibodies That Neutralize SARS-CoV-2 and Its Variants of Concern Alpha, Beta, Gamma and Delta by Binding Conformational Epitopes of Glycosylated RBD With High Potency.
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Mariotti S, Capocefalo A, Chiantore MV, Iacobino A, Teloni R, De Angelis ML, Gallinaro A, Pirillo MF, Borghi M, Canitano A, Michelini Z, Baggieri M, Marchi A, Bucci P, McKay PF, Acchioni C, Sandini S, Sgarbanti M, Tosini F, Di Virgilio A, Venturi G, Marino F, Esposito V, Di Bonito P, Magurano F, Cara A, Negri D, and Nisini R
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- Angiotensin-Converting Enzyme 2 genetics, Animals, Binding Sites, Antibody immunology, Cell Line, Tumor, Chlorocebus aethiops, Female, Glycosylation, HEK293 Cells, Humans, Mice, Inbred BALB C, Neutralization Tests, SARS-CoV-2 immunology, Spike Glycoprotein, Coronavirus genetics, Vero Cells, COVID-19 Drug Treatment, Mice, Antibodies, Monoclonal pharmacology, Antibodies, Neutralizing pharmacology, Antibodies, Viral pharmacology, Epitopes immunology, SARS-CoV-2 drug effects, Spike Glycoprotein, Coronavirus immunology
- Abstract
Antibodies targeting Receptor Binding Domain (RBD) of SARS-CoV-2 have been suggested to account for the majority of neutralizing activity in COVID-19 convalescent sera and several neutralizing antibodies (nAbs) have been isolated, characterized and proposed as emergency therapeutics in the form of monoclonal antibodies (mAbs). However, SARS-CoV-2 variants are rapidly spreading worldwide from the sites of initial identification. The variants of concern (VOC) B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma) and B.1.167.2 (Delta) showed mutations in the SARS-CoV-2 spike protein potentially able to cause escape from nAb responses with a consequent reduction of efficacy of vaccines and mAbs-based therapy. We produced the recombinant RBD (rRBD) of SARS-CoV-2 spike glycoprotein from the Wuhan-Hu 1 reference sequence in a mammalian system, for mice immunization to isolate new mAbs with neutralizing activity. Here we describe four mAbs that were able to bind the rRBD in Enzyme-Linked Immunosorbent Assay and the transmembrane full-length spike protein expressed in HEK293T cells by flow cytometry assay. Moreover, the mAbs recognized the RBD in supernatants of SARS-CoV-2 infected VERO E6 cells by Western Blot under non-reducing condition or in supernatants of cells infected with lentivirus pseudotyped for spike protein, by immunoprecipitation assay. Three out of four mAbs lost their binding efficiency to completely N-deglycosylated rRBD and none was able to bind the same recombinant protein expressed in Escherichia coli , suggesting that the epitopes recognized by three mAbs are generated by the conformational structure of the glycosylated native protein. Of particular relevance, three mAbs were able to inhibit Wuhan SARS-CoV-2 infection of VERO E6 cells in a plaque-reduction neutralization test and the Wuhan SARS-CoV-2 as well as the Alpha, Beta, Gamma and Delta VOC in a pseudoviruses-based neutralization test. These mAbs represent important additional tools for diagnosis and therapy of COVID-19 and may contribute to the understanding of the functional structure of SARS-CoV-2 RBD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mariotti, Capocefalo, Chiantore, Iacobino, Teloni, De Angelis, Gallinaro, Pirillo, Borghi, Canitano, Michelini, Baggieri, Marchi, Bucci, McKay, Acchioni, Sandini, Sgarbanti, Tosini, Di Virgilio, Venturi, Marino, Esposito, Di Bonito, Magurano, Cara, Negri and Nisini.)
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- 2021
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16. Type I Interferons in COVID-19 Pathogenesis.
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Palermo E, Di Carlo D, Sgarbanti M, and Hiscott J
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Among the many activities attributed to the type I interferon (IFN) multigene family, their roles as mediators of the antiviral immune response have emerged as important components of the host response to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. Viruses likewise have evolved multiple immune evasion strategies to circumvent the host immune response and promote virus propagation and dissemination. Therefore, a thorough characterization of host-virus interactions is essential to understand SARS-CoV-2 pathogenesis. Here, we summarize the virus-mediated evasion of the IFN responses and the viral functions involved, the genetic basis of IFN production in SARS-CoV-2 infection and the progress of clinical trials designed to utilize type I IFN as a potential therapeutic tool.
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- 2021
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17. Risk of Atypical HUS Among Family Members of Patients Carrying Complement Regulatory Gene Abnormality.
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Ardissino G, Longhi S, Porcaro L, Pintarelli G, Strumbo B, Capone V, Cresseri D, Loffredo G, Tel F, Salardi S, Sgarbanti M, Martelli L, Rodrigues EM, Borsa-Ghiringhelli N, Montini G, Seia M, Cugno M, Carfagna F, Consonni D, and Tedeschi S
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Introduction: Atypical hemolytic uremic syndrome (aHUS) is mainly due to complement regulatory gene abnormalities with a dominant pattern but incomplete penetrance. Thus, healthy carriers can be identified in any family of aHUS patients, but it is unpredictable if they will eventually develop aHUS., Methods: Patients are screened for 10 complement regulatory gene abnormalities and once a genetic alteration is identified, the search is extended to at-risk family members. The present cohort study includes 257 subjects from 71 families: 99 aHUS patients (71 index cases + 28 affected family members) and 158 healthy relatives with a documented complement gene abnormality., Results: Fourteen families (19.7%) experienced multiple cases. Over a cumulative observation period of 7595 person-years, only 28 family members carrying gene mutations experienced aHUS (overall penetrance of 20%), leading to a disease rate of 3.69 events for 1000 person-years. The disease rate was 7.47 per 1000 person-years among siblings, 6.29 among offspring, 2.01 among parents, 1.84 among carriers of variants of uncertain significance, and 4.43 among carriers of causative variants., Conclusions: The penetrance of aHUS seems a lot lower than previously reported. Moreover, the disease risk is higher in carriers of causative variants and is not equally distributed among generations: siblings and the offspring of patients have a much greater disease risk than parents. However, risk calculation may depend on variant classification that could change over time., (© 2021 International Society of Nephrology. Published by Elsevier Inc.)
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- 2021
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18. Systematic review of clinical practice guidelines for adults with fractures: identification of best evidence for rehabilitation to develop the WHO's Package of Interventions for Rehabilitation.
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Gimigliano F, Liguori S, Moretti A, Toro G, Rauch A, Negrini S, Curci C, Patrini M, Peschi L, Pournajaf S, Sgarbanti M, and Iolascon G
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- Adult, Femoral Fractures therapy, Hip Fractures rehabilitation, Hip Fractures therapy, Humans, Radius Fractures therapy, Recovery of Function, World Health Organization, Femoral Fractures rehabilitation, Practice Guidelines as Topic standards, Radius Fractures rehabilitation
- Abstract
Background: The identification of existing rehabilitation interventions and related evidence represents a crucial step along the development of the World Health Organization's (WHO) Package of Interventions for Rehabilitation (PIR). The methods for such identification have been developed by the WHO Rehabilitation Programme and Cochrane Rehabilitation under the guidance of the WHO's Guideline Review Committee secretariat. The aim of this paper is to report on the results of the systematic search for clinical practice guidelines (CPGs) relevant to the rehabilitation of adults with fractures and to present the current state of evidence available from the identified CPGs., Methods: This paper is part of the Best Evidence for Rehabilitation (be4rehab) series, developed according to the methodology presented in the World Health Organization's (WHO) Package of Interventions for Rehabilitation (PIR) introductory paper. It is a systematic review of existing CPGs on fractures in adult population published from 2009 to 2019., Results: We identified 23 relevant CPGs after title and abstract screening. According to inclusion/exclusion criteria, we selected 13 CPGs. After checking for quality, publication time, multiprofessionality, and comprehensiveness, we finally included five CPGs dealing with rehabilitative management of fractures in adult population, two CPGs addressing treatment of distal radius fracture and three the treatment of femoral/hip fracture., Conclusion: The selected CPGs on management of distal radius and femoral/hip fracture include few recommendations regarding rehabilitation, with overall low to very low quality of evidence and weak/conditional strength of recommendation. Moreover, several gaps in specific rehabilitative topics occur. Further high-quality trials are required to upgrade the quality of the available evidence., Level of Evidence: Level 1.
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- 2020
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19. IκB kinase-ε-mediated phosphorylation triggers IRF-1 degradation in breast cancer cells.
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Remoli AL, Sgarbanti M, Perrotti E, Acchioni M, Orsatti R, Acchioni C, Battistini A, Clarke R, and Marsili G
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- Apoptosis, Breast Neoplasms genetics, Breast Neoplasms metabolism, Cell Proliferation, Female, Humans, I-kappa B Kinase genetics, Interferon Regulatory Factor-1 genetics, Phosphorylation, Proteolysis, Signal Transduction, Tumor Cells, Cultured, Ubiquitination, Breast Neoplasms pathology, I-kappa B Kinase metabolism, Interferon Regulatory Factor-1 metabolism, Ubiquitin metabolism
- Abstract
Interferon Regulatory Factors (IRFs) are key regulators of immunity, cell survival and apoptosis. IRF transcriptional activity and subcellular localization are tightly regulated by posttranscriptional modifications including phosphorylation. The IκB kinase family member IKK-ε is essential in regulating antiviral innate immunity mediated by IRFs but is now also recognized as an oncoprotein amplified and overexpressed in breast cancer cell lines and patient-derived tumors. In the present study, we report that the tumor suppressor IRF-1 is a specific target of IKK-ε in breast cancer cells. IKK-ε-mediated phosphorylation of IRF-1 dramatically decreases IRF-1 protein stability, accelerating IRF-1 degradation and quenching IRF-1 transcriptional activity. Chemical inhibition of IKK-ε activity, fully restores IRF-1 levels and function and positively correlates with inhibition of cell growth and proliferation of breast cancer cells. By using a breast cancer cell line stably expressing a dominant negative version of IRF-1 we were able to demonstrate that IKK-ε preferentially exerts its oncogenic potential in breast cancer through the regulation of IRF-1 and point to the IKK-ε-mediated phosphorylation of IRF-1 as a therapeutic target to overcome IKK-ε-mediated tumorigenesis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2020
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20. Activation of Latent HIV-1 T Cell Reservoirs with a Combination of Innate Immune and Epigenetic Regulators.
- Author
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Palermo E, Acchioni C, Di Carlo D, Zevini A, Muscolini M, Ferrari M, Castiello L, Virtuoso S, Borsetti A, Antonelli G, Turriziani O, Sgarbanti M, and Hiscott J
- Subjects
- Gene Expression Regulation, Viral, HIV Infections genetics, HIV Infections virology, Histone Deacetylase Inhibitors pharmacology, Humans, Hydroxamic Acids pharmacology, Leukocytes, Mononuclear drug effects, Leukocytes, Mononuclear immunology, Sulfonamides pharmacology, Virus Replication, CD4-Positive T-Lymphocytes immunology, Epigenesis, Genetic, HIV Infections immunology, HIV-1 immunology, Immunity, Innate immunology, Virus Activation immunology, Virus Latency immunology
- Abstract
The presence of T cell reservoirs in which human immunodeficiency virus (HIV) establishes latency by integrating into the host genome represents a major obstacle to an HIV cure and has prompted the development of strategies aimed at the eradication of HIV from latently infected cells. The "shock-and-kill" strategy is one of the most pursued approaches to the elimination of viral reservoirs. Although several latency-reversing agents (LRAs) have shown promising reactivation activity, they have failed to eliminate the cellular reservoir. In this study, we evaluated a novel immune system-mediated approach to clearing the HIV reservoir, based on a combination of innate immune stimulation and epigenetic reprogramming. The combination of the STING agonist cGAMP (cyclic GMP-AMP) and the FDA-approved histone deacetylase inhibitor resminostat resulted in a significant increase in HIV proviral reactivation and specific apoptosis in HIV-infected cells in vitro Reductions in the proportion of HIV-harboring cells and the total amount of HIV DNA were also observed in CD4
+ central memory T (TCM ) cells, a primary cell model of latency, where resminostat alone or together with cGAMP induced high levels of selective cell death. Finally, high levels of cell-associated HIV RNA were detected ex vivo in peripheral blood mononuclear cells (PBMCs) and CD4+ T cells from individuals on suppressive antiretroviral therapy (ART). Although synergism was not detected in PBMCs with the combination, viral RNA expression was significantly increased in CD4+ T cells. Collectively, these results represent a promising step toward HIV eradication by demonstrating the potential of innate immune activation and epigenetic modulation for reducing the viral reservoir and inducing specific death of HIV-infected cells. IMPORTANCE One of the challenges associated with HIV-1 infection is that despite antiretroviral therapies that reduce HIV-1 loads to undetectable levels, proviral DNA remains dormant in a subpopulation of T lymphocytes. Numerous strategies to clear residual virus by reactivating latent virus and eliminating the reservoir of HIV-1 (so-called "shock-and-kill" strategies) have been proposed. In the present study, we use a combination of small molecules that activate the cGAS-STING antiviral innate immune response (the di-cyclic nucleotide cGAMP) and epigenetic modulators (histone deacetylase inhibitors) that induce reactivation and HIV-infected T cell killing in cell lines, primary T lymphocytes, and patient samples. These studies represent a novel strategy for HIV eradication by reducing the viral reservoir and inducing specific death of HIV-infected cells., (Copyright © 2019 American Society for Microbiology.)- Published
- 2019
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21. A model of the three-dimensional structure of human interferon responsive factor 1 and its modifications upon phosphorylation or phosphorylation-mimicking mutations.
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Storchi L, Remoli AL, Marsili G, Acchioni C, Acchioni M, Battistini A, Sgarbanti M, and Marrone A
- Subjects
- Aspartic Acid genetics, HEK293 Cells, Humans, Interferon Regulatory Factor-1 metabolism, Interferon-beta metabolism, Molecular Dynamics Simulation, Mutant Proteins chemistry, Phosphorylation, Static Electricity, Tumor Necrosis Factor-alpha metabolism, Interferon Regulatory Factor-1 chemistry, Interferon Regulatory Factor-1 genetics, Models, Molecular, Mutation genetics
- Abstract
Interferon responsive factor 1 (IRF-1) is a pleiotropic transcription factor, possessing non-redundant biological activities that depend on its interaction with different protein partners and multiple post-translational modifications including phosphorylation. In particular, a 5'-SXXXSXS-3' motif of the protein represents the target of the I κ B-related kinases, TANK-binding kinase (TBK)-1 and inhibitor of nuclear factor kappa-B kinase (IKK)-ε. Here, a 3D model of human IRF-1 was determined by using multi-template comparative modeling and molecular dynamics approaches. Models obtained through either phosphorylation or aspartate mutation of residues 215, 219 and 221 were also calculated and compared to the wild type. Calculations indicated that each of these modifications mainly induces a rigidification of the protein structure and only slightly changes in electrostatics and hydrophobicity of IRF-1 surface, resulting in the impairment of the capacity of IRF-1 containing as partate mutations (S221D and S215D/S219D/S221D) to synergize with tumour necrosis factor (TNF)-α stimulation in inducing interferon (IFN) promoter-mediated reporter gene activation. Therefore, these changes are qualitatively correlated to the amount of negative charge located on the 215-221 segments of IRF-1 by phosphorylation or aspartate mutation. Hypotheses on the structural mechanism that governs the phosphorylation-related damping of IRF-1 activity were also drawn. Communicated by Ramaswamy H. Sarma.
- Published
- 2019
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22. Alternate NF-κB-Independent Signaling Reactivation of Latent HIV-1 Provirus.
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Acchioni C, Remoli AL, Marsili G, Acchioni M, Nardolillo I, Orsatti R, Farcomeni S, Palermo E, Perrotti E, Barreca ML, Sabatini S, Sandini S, Parolin C, Lin R, Borsetti A, Hiscott J, and Sgarbanti M
- Subjects
- Anti-Retroviral Agents therapeutic use, CD4-Positive T-Lymphocytes immunology, Gene Expression Regulation, Viral genetics, HIV Infections virology, HIV Seropositivity immunology, HIV-1 physiology, Humans, Jurkat Cells, NF-KappaB Inhibitor alpha metabolism, NF-kappa B metabolism, Proviruses drug effects, Proviruses physiology, Receptors, Immunologic metabolism, Signal Transduction drug effects, Transcription Factor RelA metabolism, Virus Replication drug effects, HIV-1 drug effects, Virus Activation drug effects, Virus Latency drug effects
- Abstract
Current combination antiretroviral therapies (cART) are unable to eradicate HIV-1 from infected individuals because of the establishment of proviral latency in long-lived cellular reservoirs. The shock-and-kill approach aims to reactivate viral replication from the latent state (shock) using latency-reversing agents (LRAs), followed by the elimination of reactivated virus-producing cells (kill) by specific therapeutics. The NF-κB RelA/p50 heterodimer has been characterized as an essential component of reactivation of the latent HIV-1 long terminal repeat (LTR). Nevertheless, prolonged NF-κB activation contributes to the development of various autoimmune, inflammatory, and malignant disorders. In the present study, we established a cellular model of HIV-1 latency in J-Lat CD4
+ T cells that stably expressed the NF-κB superrepressor IκB-α 2NΔ4 and demonstrate that conventional treatments with bryostatin-1 and hexamethylenebisacetamide (HMBA) or ionomycin synergistically reactivated HIV-1 from latency, even under conditions where NF-κB activation was repressed. Using specific calcineurin phosphatase, p38, and MEK1/MEK2 kinase inhibitors or specific short hairpin RNAs, c-Jun was identified to be an essential factor binding to the LTR enhancer κB sites and mediating the combined synergistic reactivation effect. Furthermore, acetylsalicylic acid (ASA), a potent inhibitor of the NF-κB activator kinase IκB kinase β (IKK-β), did not significantly diminish reactivation in a primary CD4+ T central memory (TCM ) cell latency model. The present work demonstrates that the shock phase of the shock-and-kill approach to reverse HIV-1 latency may be achieved in the absence of NF-κB, with the potential to avoid unwanted autoimmune- and or inflammation-related side effects associated with latency-reversing strategies. IMPORTANCE The shock-and-kill approach consists of the reactivation of HIV-1 replication from latency using latency-reversing agents (LRAs), followed by the elimination of reactivated virus-producing cells. The cellular transcription factor NF-κB is considered a master mediator of HIV-1 escape from latency induced by LRAs. Nevertheless, a systemic activation of NF-κB in HIV-1-infected patients resulting from the combined administration of different LRAs could represent a potential risk, especially in the case of a prolonged treatment. We demonstrate here that conventional treatments with bryostatin-1 and hexamethylenebisacetamide (HMBA) or ionomycin synergistically reactivate HIV-1 from latency, even under conditions where NF-κB activation is repressed. Our study provides a molecular proof of concept for the use of anti-inflammatory drugs, like aspirin, capable of inhibiting NF-κB in patients under combination antiretroviral therapy during the shock-and-kill approach, to avoid potential autoimmune and inflammatory disorders that can be elicited by combinations of LRAs., (Copyright © 2019 American Society for Microbiology.)- Published
- 2019
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23. Complement functional tests for monitoring eculizumab treatment in patients with atypical hemolytic uremic syndrome: an update.
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Ardissino G, Tel F, Sgarbanti M, Cresseri D, Giussani A, Griffini S, Grovetto E, Possenti I, Perrone M, Testa S, Paglialonga F, Messa P, and Cugno M
- Subjects
- Adolescent, Adult, Child, Child, Preschool, Complement System Proteins analysis, Female, Humans, Infant, Male, Middle Aged, Retrospective Studies, Young Adult, Antibodies, Monoclonal, Humanized therapeutic use, Atypical Hemolytic Uremic Syndrome drug therapy, Complement Inactivating Agents therapeutic use, Complement System Proteins drug effects, Drug Monitoring methods
- Abstract
Background: Atypical hemolytic uremic syndrome (aHUS) is a thrombotic microangiopathy (TMA) characterized by platelet consumption, hemolysis, and organ damage. Eculizumab (ECU), a humanized antibody that blocks complement activity, has been successfully used in aHUS, but the best treatment schedule is not yet clear., Methods: Here, we report our experience with ECU maintenance treatment and the interval between subsequent doses being extended based on global classical complement pathway (CCP) activity aimed at <30% for maintaining aHUS into remission., Results: We report on 38 patients with aHUS, 13 children, 21 female, with a median age of 25.0 years (range 0.5-60) at disease onset treated with ECU standard schedule for a median of 2.6 months (range 0.4-24.6). Once stable TMA remission was obtained, the interval between ECU doses was extended based on complement function, with a target CCP activity of <30%. With this approach, 22 patients regularly receive ECU infusion every 28 days and 16 every 21. During a median observation period on ECU, an extended interval of 26.9 months (range 0.8-80.9), with a cumulative observation period of 1,208 months, none of the patients relapsed., Conclusion: Monitoring complement activity allows a safe reduction in the frequency of ECU administration in aHUS while keeping the disease in remission.
- Published
- 2018
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24. Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling.
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Fanunza E, Frau A, Sgarbanti M, Orsatti R, Corona A, and Tramontano E
- Subjects
- Drug Discovery, HEK293 Cells, Host-Pathogen Interactions drug effects, Humans, Interferon-alpha pharmacology, Promoter Regions, Genetic drug effects, Reproducibility of Results, Signal Transduction drug effects, Transfection, Viral Proteins drug effects, Ebolavirus genetics, Genes, Reporter genetics, Interferon-beta genetics, Luciferases genetics, Viral Proteins genetics, Viral Proteins metabolism
- Abstract
The interferon (IFN) system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24) is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs) and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T) cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE). Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z'- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance., Competing Interests: The authors declare no conflict of interest.
- Published
- 2018
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25. [Complement factor B mutation in atypical hemolytic uremic syndrome. Rare cause of rare disease].
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Visconti L, Cernaro V, Ardissino G, Sgarbanti M, Ferrara D, Visconti G, Santoro D, and Buemi M
- Subjects
- Aged, Humans, Male, Atypical Hemolytic Uremic Syndrome genetics, Complement Factor B genetics, Mutation, Rare Diseases genetics
- Abstract
Hemolytic uremic syndrome (HUS) is a rare disease characterized by microangiopathic hemolysis, platelet consumption and multiple organ failure with predominant renal involvement. In the most of cases (85-90%), it is associated with enteric infection due to Shiga-toxin or verocytotoxin (STEC-VTEC)-producer Escherichia coli. Rarely, in about 10-15% of cases, HUS develops in the presence of a disorder of alternative complement pathway regulation and it is defined atypical (aHUS). We describe the case of a 65-year-old man who came to our attention with a clinical presentation of aHUS and a clinical course characterized by rapidly progressive acute renal failure (ARF), which required renal replacement treatments, and by a stable clinical picture of hematological impairment as a marker of a non-severe and self-limiting form. The clinical and laboratory course allowed us not to perform specific therapies such as plasma exchange and/or block of the complement with eculizumab. Less than two weeks after hospital admission, there was a gradual recovery of renal function with spontaneous diuresis and hematological remission. Genetic screening has revealed a heterozygous mutation in the complement factor B (CFB) that is not described in the literature and therefore not yet characterized in the genotype/phenotype correlation, also for the extreme rarity of the forms associated with CFB alteration. In conclusion, the presence of a new mutation in the CFB, such as the one described in our case, is probably associated with the development of aHUS but has not led to a poor prognosis, as generally reported in the literature for known variants of the CFB., (Copyright by Società Italiana di Nefrologia SIN, Rome, Italy.)
- Published
- 2017
26. HIV-1 Tat Recruits HDM2 E3 Ligase To Target IRF-1 for Ubiquitination and Proteasomal Degradation.
- Author
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Remoli AL, Marsili G, Perrotti E, Acchioni C, Sgarbanti M, Borsetti A, Hiscott J, and Battistini A
- Subjects
- Cell Line, Humans, Protein Binding, Ubiquitination, HIV-1 pathogenicity, Host-Pathogen Interactions, Interferon Regulatory Factor-1 metabolism, Protein Processing, Post-Translational, Proto-Oncogene Proteins c-mdm2 metabolism, tat Gene Products, Human Immunodeficiency Virus metabolism
- Abstract
In addition to its ability to regulate HIV-1 promoter activation, the viral transactivator Tat also functions as a determinant of pathogenesis and disease progression by directly and indirectly modulating the host anti-HIV response, largely through the capacity of Tat to interact with and modulate the activities of multiple host proteins. We previously demonstrated that Tat modulated both viral and host transcriptional machinery by interacting with the cellular transcription factor interferon regulatory factor 1 (IRF-1). In the present study, we investigated the mechanistic basis and functional significance of Tat-IRF-1 interaction and demonstrate that Tat dramatically decreased IRF-1 protein stability. To accomplish this, Tat exploited the cellular HDM2 (human double minute 2 protein) ubiquitin ligase to accelerate IRF-1 proteasome-mediated degradation, resulting in a quenching of IRF-1 transcriptional activity during HIV-1 infection. These data identify IRF-1 as a new target of Tat-induced modulation of the cellular protein machinery and reveal a new strategy developed by HIV-1 to evade host immune responses., Importance: Current therapies have dramatically reduced morbidity and mortality associated with HIV infection and have converted infection from a fatal pathology to a chronic disease that is manageable via antiretroviral therapy. Nevertheless, HIV-1 infection remains a challenge, and the identification of useful cellular targets for therapeutic intervention remains a major goal. The cellular transcription factor IRF-1 impacts various physiological functions, including the immune response to viral infection. In this study, we have identified a unique mechanism by which HIV-1 evades IRF-1-mediated host immune responses and show that the viral protein Tat accelerates IRF-1 proteasome-mediated degradation and inactivates IRF-1 function. Restoration of IRF-1 functionality may thus be regarded as a potential strategy to reinstate both a direct antiviral response and a more broadly acting immune regulatory circuit., (Copyright © 2016 Remoli et al.)
- Published
- 2016
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27. IFN Regulatory Factors and Antiviral Innate Immunity: How Viruses Can Get Better.
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Marsili G, Perrotti E, Remoli AL, Acchioni C, Sgarbanti M, and Battistini A
- Subjects
- Animals, Gene Expression Regulation, Host-Pathogen Interactions genetics, Humans, Interferon Regulatory Factors genetics, Multigene Family, Signal Transduction, Virus Diseases genetics, Virus Diseases virology, Host-Pathogen Interactions immunology, Immunity, Innate, Interferon Regulatory Factors metabolism, Virus Diseases immunology, Virus Diseases metabolism, Viruses immunology
- Abstract
The interferon regulatory factor (IRF) family consists of transcriptional regulators that exert multifaceted and versatile functions in multiple biological processes. Their crucial role as central mediators in the establishment and execution of host immunity in response to pathogen-derived signals downstream pattern recognition receptors (PRRs) makes IRFs a hallmark of the host antiviral response. They function as hub molecules at the crossroad of different signaling pathways for the induction of interferon (IFN) and inflammatory cytokines, as well as of antiviral and immunomodulatory genes even in an IFN-independent manner. By regulating the development and activity of immune cells, IRFs also function as a bridge between innate and adaptive responses. As such, IRFs represent attractive and compulsive targets in viral strategies to subvert antiviral signaling. In this study, we discuss current knowledge on the wide array of strategies put in place by pathogenic viruses to evade, subvert, and/or hijack these essential components of host antiviral immunity.
- Published
- 2016
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28. Type I IFN--a blunt spear in fighting HIV-1 infection.
- Author
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Acchioni C, Marsili G, Perrotti E, Remoli AL, Sgarbanti M, and Battistini A
- Subjects
- Disease Progression, HIV Infections complications, HIV Infections virology, Humans, Immune Evasion, Immunity, Innate, Inflammation, Interferon Regulatory Factors immunology, Interferon Type I biosynthesis, Pathogen-Associated Molecular Pattern Molecules metabolism, Viral Proteins immunology, Virus Replication, HIV Infections immunology, HIV-1 immunology, HIV-1 pathogenicity, Interferon Type I immunology
- Abstract
For more than 50 years, Type I Interferon (IFN) has been recognized as critical in controlling viral infections. IFN is produced downstream germ-line encoded pattern recognition receptors (PRRs) upon engagement by pathogen-associated molecular patterns (PAMPs). As a result, hundreds of different interferon-stimulated genes (ISGs) are rapidly induced, acting in both autocrine and paracrine manner to build a barrier against viral replication and spread. ISGs encode proteins with direct antiviral and immunomodulatory activities affecting both innate and adaptive immune responses. During infection with viruses, as HIV-1, that can establish a persistent infection, IFN although produced, is not able to block the initial infection and a chronic IFN-mediated immune activation/inflammation becomes a pathogenic mechanism of disease progression. This review will briefly summarize when and how IFN is produced during HIV-1 infection and the way this innate immune response is manipulated by the virus to its own advantage to drive chronic immune activation and progression to AIDS., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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29. HIV-1 latency: an update of molecular mechanisms and therapeutic strategies.
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Battistini A and Sgarbanti M
- Subjects
- Anti-HIV Agents isolation & purification, Drug Discovery trends, Drug Therapy methods, HIV Infections virology, Humans, Anti-HIV Agents pharmacology, Anti-HIV Agents therapeutic use, HIV Infections drug therapy, HIV-1 drug effects, HIV-1 physiology, Virus Latency drug effects
- Abstract
The major obstacle towards HIV-1 eradication is the life-long persistence of the virus in reservoirs of latently infected cells. In these cells the proviral DNA is integrated in the host's genome but it does not actively replicate, becoming invisible to the host immune system and unaffected by existing antiviral drugs. Rebound of viremia and recovery of systemic infection that follows interruption of therapy, necessitates life-long treatments with problems of compliance, toxicity, and untenable costs, especially in developing countries where the infection hits worst. Extensive research efforts have led to the proposal and preliminary testing of several anti-latency compounds, however, overall, eradication strategies have had, so far, limited clinical success while posing several risks for patients. This review will briefly summarize the more recent advances in the elucidation of mechanisms that regulates the establishment/maintenance of latency and therapeutic strategies currently under evaluation in order to eradicate HIV persistence.
- Published
- 2014
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30. IκB kinase ε targets interferon regulatory factor 1 in activated T lymphocytes.
- Author
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Sgarbanti M, Marsili G, Remoli AL, Stellacci E, Mai A, Rotili D, Perrotti E, Acchioni C, Orsatti R, Iraci N, Ferrari M, Borsetti A, Hiscott J, and Battistini A
- Subjects
- Acetylation, CD28 Antigens genetics, CD28 Antigens metabolism, CD3 Complex genetics, CD3 Complex metabolism, Cell Line, HEK293 Cells, Humans, I-kappa B Kinase metabolism, Interferon Regulatory Factor-1 metabolism, Interferon-beta genetics, Interferon-beta metabolism, Protein Processing, Post-Translational genetics, Transcription Factor RelA genetics, Transcription Factor RelA metabolism, Transcription, Genetic genetics, p300-CBP Transcription Factors genetics, p300-CBP Transcription Factors metabolism, CD4-Positive T-Lymphocytes metabolism, I-kappa B Kinase genetics, Interferon Regulatory Factor-1 genetics, Lymphocyte Activation genetics
- Abstract
IκB kinase ε (IKK-ε) has an essential role as a regulator of innate immunity, functioning downstream of pattern recognition receptors to modulate NF-κB and interferon (IFN) signaling. In the present study, we investigated IKK-ε activation following T cell receptor (TCR)/CD28 stimulation of primary CD4(+) T cells and its role in the stimulation of a type I IFN response. IKK-ε was activated following TCR/CD28 stimulation of primary CD4(+) T cells; however, in T cells treated with poly(I·C), TCR/CD28 costimulation blocked induction of IFN-β transcription. We demonstrated that IKK-ε phosphorylated the transcription factor IFN regulatory factor 1 (IRF-1) at amino acid (aa) 215/219/221 in primary CD4(+) T cells and blocked its transcriptional activity. At the mechanistic level, IRF-1 phosphorylation impaired the physical interaction between IRF-1 and the NF-κB RelA subunit and interfered with PCAF-mediated acetylation of NF-κB RelA. These results demonstrate that TCR/CD28 stimulation of primary T cells stimulates IKK-ε activation, which in turn contributes to suppression of IFN-β production.
- Published
- 2014
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31. Therapeutics for HIV-1 reactivation from latency.
- Author
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Sgarbanti M and Battistini A
- Subjects
- Animals, HIV Infections virology, HIV-1 drug effects, HIV-1 genetics, Humans, Anti-HIV Agents therapeutic use, HIV Infections drug therapy, HIV-1 physiology, Virus Activation drug effects, Virus Latency drug effects
- Abstract
Intensive combined antiretroviral therapy successfully suppresses HIV-1 replication and AIDS disease progression making infection manageable, but it is unable to eradicate the virus that persists in long-lived, drug-insensitive and immune system-insensitive reservoirs thus asking for life-long treatments with problems of compliance, resistance, toxicity and cost. These limitations and recent insights into latency mechanisms have fueled a renewed effort in finding a cure for HIV-1 infection. Proposed eradication strategies involve reactivation of the latent reservoir upon induction of viral transcription followed by the elimination of reactivated virus-producing cells by viral cytopathic effect or host immune response. Several molecules identified by mechanism-directed approaches or in large-scale screenings have been proposed as latency reversing agents. Some of them have already entered clinical testing in humans but with mixed or unsatisfactory results., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2013
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32. HIV-1, interferon and the interferon regulatory factor system: an interplay between induction, antiviral responses and viral evasion.
- Author
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Marsili G, Remoli AL, Sgarbanti M, Perrotti E, Fragale A, and Battistini A
- Subjects
- HIV Infections metabolism, HIV Infections virology, HIV-1 physiology, Host-Pathogen Interactions immunology, Humans, Immune Evasion immunology, Immunity, Innate immunology, Interferon Regulatory Factors metabolism, Interferons metabolism, Models, Immunological, Signal Transduction immunology, HIV Infections immunology, HIV-1 immunology, Interferon Regulatory Factors immunology, Interferons immunology
- Abstract
Thirty years after the first isolation of the etiological agent of AIDS, the virus HIV-1 is still a major threat worldwide with millions of individuals currently infected. Although current combination therapies allow viral replication to be controlled, HIV-1 is not eradicated and persists in drug- and immune system-insensitive reservoirs and a cure is still lacking. Pathogens such as HIV-1 that cause chronic infections are able to adapt to the host in a manner that ensures long term residence and survival, via the evolution of numerous mechanisms that evade various aspects of the innate and adaptive immune response. One such mechanism is targeted to members of the interferon (IFN) regulatory factor (IRF) family of proteins. These transcription factors regulate a variety of biological processes including interferon induction, immune cell activation and downstream pattern recognition receptors (PRRs). HIV-1 renders IRFs harmless and hijacks them to its own advantage in order to facilitate its replication and evasion of immune responses. Type I interferon (IFN), the canonical antiviral innate response, can be induced in both acute and chronic HIV-1 infection in vivo, but in the majority of individuals this initial response is not protective and can contribute to disease progression. Type I IFN expression is largely inhibited in T cells and macrophages in order to successfully establish productive infection, whereas sustained IFN production by plasmacytoid dendritic cells is considered an important source of chronic immune activation, a hallmark to AIDS progression., (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Published
- 2012
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33. The development of immune-modulating compounds to disrupt HIV latency.
- Author
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Remoli AL, Marsili G, Battistini A, and Sgarbanti M
- Subjects
- Anti-HIV Agents immunology, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes virology, HIV Infections immunology, HIV Infections virology, HIV-1 immunology, HIV-1 physiology, Host-Pathogen Interactions drug effects, Host-Pathogen Interactions immunology, Humans, Immunologic Factors immunology, Models, Immunological, Virus Activation drug effects, Virus Activation immunology, Virus Latency immunology, Anti-HIV Agents therapeutic use, HIV Infections prevention & control, HIV-1 drug effects, Immunologic Factors therapeutic use, Virus Latency drug effects
- Abstract
Antiretroviral therapy (ART) has proved highly effective in suppressing HIV-1 replication and disease progression. Nevertheless, ART has failed to eliminate the virus from infected individuals. The main obstacle to HIV-1 eradication is the persistence of cellular viral reservoirs. Therefore, the "shock-and-kill" strategy was proposed consisting of inducing HIV-1 escape from latency, in the presence of ART. This is followed by the elimination of reactivated, virus-producing cells. Immune modulators, including protein kinase C (PKC) activators, anti-leukemic drugs and histone deacetylase inhibitors (HDACis) have all demonstrated efficacy in the reactivation of latent virus replication. This review will focus on the potential use of these small molecules in the "shock and kill" strategy, the molecular basis for their action and the potential advantages of their immune-modulating activities., (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Published
- 2012
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34. The design of optimal therapeutic small interfering RNA molecules targeting diverse strains of influenza A virus.
- Author
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ElHefnawi M, Hassan N, Kamar M, Siam R, Remoli AL, El-Azab I, AlAidy O, Marsili G, and Sgarbanti M
- Subjects
- HEK293 Cells, Humans, Influenza A Virus, H5N1 Subtype genetics, Influenza A Virus, H5N1 Subtype physiology, Influenza, Human genetics, Software, Influenza A virus classification, Influenza A virus genetics, Influenza, Human therapy, RNA, Small Interfering genetics
- Abstract
Motivation: There is an urgent need for new medications to combat influenza pandemics., Methods: Using the genome analysis of the influenza A virus performed previously, we designed and performed a combinatorial exhaustive systematic methodology for optimal design of universal therapeutic small interfering RNA molecules (siRNAs) targeting all diverse influenza A viral strains. The rationale was to integrate the factors for highly efficient design in a pipeline of analysis performed on possible influenza-targeting siRNAs. This analysis selects specific siRNAs that has the ability to target highly conserved, accessible and biologically significant regions. This would require minimal dosage and side effects., Results and Discussion: First, >6000 possible siRNAs were designed. Successive filtration followed where a novel method for siRNA scoring filtration layers was implemented. This method excluded siRNAs below the 90% experimental inhibition mapped scores using the intersection of 12 different scoring algorithms. Further filtration of siRNAs is done by eliminating those with off-targets in the human genome and those with undesirable properties and selecting siRNA targeting highly probable single-stranded regions. Finally, the optimal properties of the siRNA were ensured through selection of those targeting 100% conserved, biologically functional short motifs. Validation of a predicted active (sh114) and a predicted inactive (sh113) (that was filtered out in Stage 8) silencer of the NS1 gene showed significant inhibition of the NS1 gene for sh114, with negligible decrease for sh113 which failed target accessibility. This demonstrated the fertility of this methodology., Contact: mahef@aucegypt.edu, Supplementary Information: Supplementary data are available at Bioinformatics online.
- Published
- 2011
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35. Human papillomavirus type 16 E5 protein induces expression of beta interferon through interferon regulatory factor 1 in human keratinocytes.
- Author
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Muto V, Stellacci E, Lamberti AG, Perrotti E, Carrabba A, Matera G, Sgarbanti M, Battistini A, Liberto MC, and Focà A
- Subjects
- Cell Line, Human papillomavirus 16 pathogenicity, Humans, Keratinocytes virology, Host-Pathogen Interactions, Human papillomavirus 16 immunology, Interferon Regulatory Factor-1 metabolism, Interferon-beta biosynthesis, Keratinocytes immunology, Oncogene Proteins, Viral metabolism
- Abstract
Crucial steps in high-risk human papillomavirus (HR-HPV)-related carcinogenesis are the integration of HR-HPV into the host genome and loss of viral episomes. The mechanisms that promote cervical neoplastic progression are, however, not clearly understood. During HR-HPV infection, the HPV E5 protein is expressed in precancerous stages but not after viral integration. Given that it has been reported that loss of HPV16 episomes and cervical tumor progression are associated with increased expression of antiviral genes that are inducible by type I interferon (IFN), we asked whether E5, expressed in early phases of cervical carcinogenesis, affects IFN-β signaling. We show that the HPV type 16 (HPV16) E5 protein expression per se stimulates IFN-β expression. This stimulation is specifically mediated by the induction of interferon regulatory factor 1 (IRF-1) which, in turn, induces transcriptional activation of IRF-1-targeted interferon-stimulated genes (ISGs) as double-stranded RNA-dependent protein kinase R (PKR) and caspase 8. Our data show a new and unexpected role for HR-HPV E5 protein and indicate that HPV16 E5 may contribute to the mechanisms responsible for cervical carcinogenesis in part via stimulation of IFN-β and an IFN signature, with IRF-1 playing a pivotal role. HPV16 E5 and IRF-1 may thus serve as potential therapeutic targets in HPV-associated premalignant lesions.
- Published
- 2011
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36. Interferon regulatory factor-1 acts as a powerful adjuvant in tat DNA based vaccination.
- Author
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Castaldello A, Sgarbanti M, Marsili G, Brocca-Cofano E, Remoli AL, Caputo A, and Battistini A
- Subjects
- AIDS Vaccines immunology, Animals, Cell Line, Cysteine Endopeptidases genetics, Cysteine Endopeptidases metabolism, Female, HIV-1 immunology, Humans, Immunization, Interferon Regulatory Factor-1 genetics, Interferon Regulatory Factor-3 immunology, Interferon Regulatory Factor-7 genetics, Interferon Regulatory Factor-7 immunology, Mice, T-Lymphocytes, Cytotoxic immunology, Adjuvants, Immunologic, Interferon Regulatory Factor-1 immunology, Vaccines, DNA immunology, tat Gene Products, Human Immunodeficiency Virus
- Abstract
Genetic vaccines are safe cost-effective approaches to immunization but DNA immunization is an inefficient process. There is, therefore, a pressing need for adjuvants capable of enhancing the immunogenicity and effectiveness of these vaccines. This is particularly important for diseases for which successful vaccines are still lacking, such as cancer and infectious diseases including HIV-1/AIDS. Here we report an approach to enhance the immunogenicity of DNA vaccines involving the use of transcription factors of the Interferon regulatory factor (IRF) family, specifically IRF-1, IRF-3, and IRF-7 using the tat gene as model antigen. Balb/c mice were immunized by three intramuscular inoculations, using a DNA prime-protein boost protocol, with a DNA encoding tat of HIV-1 and the indicated IRFs and immune responses were compared to those induced by vaccination with tat DNA alone. In vivo administration of plasmid DNA encoding IRF-1, or a mutated version of IRF-1 deleted of the DNA-binding domain, enhanced Tat-specific immune responses and shifted them towards a predominant T helper 1-type immune response with increased IFN-gamma production and cytotoxic T lymphocytes responses. Conversely, the use of IRF-3 or IRF-7 did not affect the tat-induced responses. These findings define IRF-1 and its mutated form as efficacious T helper 1-inducing adjuvants in the context of tat-based vaccination and also providing a new promising candidate for genetic vaccine development., ((c) 2010 Wiley-Liss, Inc.)
- Published
- 2010
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37. Generation of a human immunodeficiency virus type 1 chronically infected monkey B cell line expressing low levels of endogenous TRIM5alpha.
- Author
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Ridolfi B, Catone S, Sgarbanti M, Sernicola L, Battistini A, Parolin C, Titti F, and Borsetti A
- Subjects
- Amino Acid Substitution genetics, Animals, Antiviral Restriction Factors, Carrier Proteins genetics, Cell Line, Cell Line, Tumor, Gene Expression genetics, HIV Core Protein p24 metabolism, HIV-1 genetics, HIV-1 ultrastructure, Human Immunodeficiency Virus Proteins genetics, Human Immunodeficiency Virus Proteins metabolism, Humans, Leukocytes, Mononuclear virology, Species Specificity, T-Lymphocytes metabolism, T-Lymphocytes virology, Tripartite Motif Proteins, Ubiquitin-Protein Ligases, Virion genetics, Virion growth & development, Virion ultrastructure, Virus Replication physiology, gag Gene Products, Human Immunodeficiency Virus metabolism, B-Lymphocytes metabolism, B-Lymphocytes virology, Carrier Proteins metabolism, HIV-1 growth & development, Macaca fascicularis
- Abstract
Several innate cellular antiviral factors exist in mammalian cells that prevent the replication of retroviruses. Among them, the tripartite motif protein (TRIM)5alpha has been shown to block human immunodeficiency virus type 1 (HIV-1) infection in several types of Old World monkey cells. Here we report a novel HIV-1 chronically infected monkey B cell line, F6/HIV-1, characterized by very low levels of TRIM5alpha expression that allows HIV-1 to overcome the restriction. Virus produced by F6/HIV-1 cells fails to infect monkey cells but retains the ability to infect human peripheral blood mononuclear cells (PBMCs) and T cell lines, although with a reduced infectivity compared to the input virus. Ultrastructural analyses revealed the presence of budding virions at the F6/HIV-1 cells plasma membrane characterized by a typical conical core shell. To our knowledge F6/HIV-1 is the first monkey cell line chronically infected by HIV-1 and able to release infectious particles thus representing a useful tool to gain further insights into the molecular mechanisms of HIV-1 pathogenesis.
- Published
- 2009
- Full Text
- View/download PDF
38. IRF-1 is required for full NF-kappaB transcriptional activity at the human immunodeficiency virus type 1 long terminal repeat enhancer.
- Author
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Sgarbanti M, Remoli AL, Marsili G, Ridolfi B, Borsetti A, Perrotti E, Orsatti R, Ilari R, Sernicola L, Stellacci E, Ensoli B, and Battistini A
- Subjects
- Binding Sites, Cell Line, Electrophoretic Mobility Shift Assay, Gene Silencing, HIV-1 genetics, Humans, Immunoprecipitation, Interferon Regulatory Factor-1 antagonists & inhibitors, Point Mutation, Protein Binding, Reverse Transcriptase Polymerase Chain Reaction, T-Lymphocytes virology, HIV Enhancer genetics, HIV Long Terminal Repeat genetics, HIV-1 physiology, Interferon Regulatory Factor-1 metabolism, NF-kappa B metabolism
- Abstract
Human immunodeficiency virus type 1 (HIV-1) gene expression is controlled by a complex interplay between viral and host factors. We have previously shown that interferon-regulatory factor 1 (IRF-1) is stimulated early after HIV-1 infection and regulates promoter transcriptional activity even in the absence of the viral transactivator Tat. In this work we demonstrate that IRF-1 is also required for full NF-kappaB transcriptional activity. We provide evidence that IRF-1 and NF-kappaB form a functional complex at the long terminal repeat (LTR) kappaB sites, which is abolished by specific mutations in the two adjacent kappaB sites in the enhancer region. Silencing IRF-1 with small interfering RNA resulted in impaired NF-kappaB-mediated transcriptional activity and in repressed HIV-1 transcription early in de novo-infected T cells. These data indicate that in early phases of HIV-1 infection or during virus reactivation from latency, when the viral transactivator is absent or present at very low levels, IRF-1 is an additional component of the p50/p65 heterodimer binding the LTR enhancer, absolutely required for efficient HIV-1 replication.
- Published
- 2008
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39. Role of acetylases and deacetylase inhibitors in IRF-1-mediated HIV-1 long terminal repeat transcription.
- Author
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Marsili G, Remoli AL, Sgarbanti M, and Battistini A
- Subjects
- Humans, Interferon Regulatory Factor-1, Jurkat Cells, Acetylesterase antagonists & inhibitors, DNA-Binding Proteins physiology, Enzyme Inhibitors pharmacology, HIV Long Terminal Repeat genetics, HIV-1 genetics, Phosphoproteins physiology, Transcription, Genetic drug effects
- Abstract
There is strong evidence that both transcriptional activation and silencing are mediated through the recruitment of enzymes that control reversible protein acetylation: histone acetylase (HAT) and histone deacetylase proteins. Acetylation is also a critical post-translational modification of general and tissue-specific transcription factors. In HIV-1-infected cells, the long terminal repeat (LTR) promoter, once organized into chromatin, is transcriptionally inactive in the absence of stimulation. LTR transcription is regulated by protein acetylation, since treatment with deacetylase inhibitors markedly induces transcriptional activity of the LTR. Besides cellular transcription factors involved in LTR activation, early in infection, and during reactivation from latency, we have previously shown that proteins of the IRF family play an important role. In particular, IRF-1 is able per se to stimulate HIV-1 LTR transcription even in the absence of Tat. IRF-1 is also acetylated and associates with HATs such as p300/CBP and PCAF to form a multiprotein complex that assembles on the promoter of target genes. Here we show that CBP can be recruited by IRF-1 to the HIV-1 LTR promoter even in the absence of Tat and that treatment with deacetylase inhibitors, such as trichostatin A (TSA), increases LTR transactivation in response to both IRF-1 and Tat. These results help to define the architecture of interactions between transcription factors binding HIV-1 LTR and confirm the possibility that deacetylase inhibitors, such as TSA, combined with antiviral therapy may represent a valuable approach to control HIV-1 infection.
- Published
- 2004
- Full Text
- View/download PDF
40. Analysis of the signal transduction pathway leading to human immunodeficiency virus-1-induced interferon regulatory factor-1 upregulation.
- Author
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Sgarbanti M, Marsili G, Remoli AL, Ridolfi B, Stellacci E, Borsetti A, Ensoli B, and Battistini A
- Subjects
- Base Sequence, Cell Line, DNA Primers, Electrophoretic Mobility Shift Assay, HIV Long Terminal Repeat, Humans, Interferon Regulatory Factor-1, Promoter Regions, Genetic, Virus Replication, DNA-Binding Proteins physiology, HIV-1 physiology, Phosphoproteins physiology, Signal Transduction, Up-Regulation physiology
- Abstract
Interferon (IFN) regulatory factors (IRFs) constitute a family of transcriptional activators and repressors involved in the regulation of immune system, host defense, and cell growth. All members share conserved DNA-binding domains that recognize DNA sequences termed IRF-binding elements/IFN-stimulated response elements (IRF-E/ISRE) present on the promoter of IFN-alpha/beta and IFN-stimulated genes. An ISRE has been identified downstream of the transcription start site of the long terminal repeat (LTR) of human immunodeficiency virus-1 (HIV-1). Our previous results showed that among the IRF factors, IRF-1 is able to stimulate HIV-1 LTR transcription and its expression is induced by HIV-1, early, upon infection and before the expression of Tat. In this study we investigated the signal transduction pathway leading to HIV-1-induced IRF-1 expression. Key IRF-1 promoter elements that mediate the activation of transcription upon induction by inflammatory cytokines are IFN-gamma-activated sequences that bind members of the signal transducer and activator of transcription (STAT) family and binding sites for nuclear factor kappaB (NF-kappaB). Both STAT-1 and NF-kappaB activation were examined to determine putative molecular targets whose inhibition resulted in the inhibition of HIV-1 replication. The results show that at early time points after HIV-1 infection, NF-kappaB but not STAT-1 is activated. Moreover, a significant decrease in HIV-1 replication was observed upon de novo infection of Jurkat T cells expressing an NF-kappaB super-repressor (IkappaB-alpha 2NDelta4). These results suggest that in early phases of HIV-1 infection, before detectable cytokine production, NF-kappaB seems responsible for HIV-1-induced IRF-1 expression.
- Published
- 2004
- Full Text
- View/download PDF
41. A requirement for NF-kappaB induction in the production of replication-competent HHV-8 virions.
- Author
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Sgarbanti M, Arguello M, tenOever BR, Battistini A, Lin R, and Hiscott J
- Subjects
- Herpesvirus 8, Human drug effects, Humans, I-kappa B Kinase, Lymphoma, B-Cell metabolism, Lymphoma, B-Cell pathology, Lymphoma, B-Cell virology, Pleural Effusion, Malignant metabolism, Pleural Effusion, Malignant pathology, Pleural Effusion, Malignant virology, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases metabolism, Tetradecanoylphorbol Acetate, Tumor Cells, Cultured, Virion drug effects, Virion metabolism, Herpesvirus 8, Human pathogenicity, Herpesvirus 8, Human physiology, NF-kappa B metabolism, Virus Replication
- Abstract
The gammaherpesvirus human herpesvirus 8 (HHV-8) infects endothelial and B-lymphoid cells and is responsible for the development of Kaposi's sarcoma and primary effusion lymphoma (PEL). In the present study, we demonstrate that the activation of the NF-kappaB pathway during HHV-8 lytic replication is required for the generation of replication-competent virions capable of initiating a de novo infection of endothelial cells. In the HHV-8-positive PEL cell line BCBL-1, tetradecanoyl phorbol acetate (TPA) induction of the lytic cycle activates the NF-kappaB pathway, and this activation requires the induction of the IKKbeta component of the classical IkappaB kinase (IKK) complex. To further investigate the role of NF-kappaB activation in HHV-8 lytic replication, the NF-kappaB super-repressor IkappaBalpha-2NDelta4 was introduced into BCBL-1 cells by retroviral transduction. Expression of IkappaBalpha-2NDelta4 completely abolished NF-kappaB activity, as demonstrated by the loss of NF-kappaB DNA-binding activity and the absence of expression of the endogenous, NF-kappaB-regulated IkappaBalpha gene. NF-kappaB blockade dramatically impaired the ability of HHV-8 to produce infectious particles capable of initiating an effective de novo infection of endothelial EA.hy926 cells, as demonstrated by the lack of viral protein production in the target cells. Diminished infectivity did not appear to be caused by a reduction in virus titer, as demonstrated by equivalent viral DNA content in the supernatant of TPA-stimulated BCBL-1 and BCBL-1/2N4 cells. Although the viral and/or cellular products affected by NF-kappaB inactivation remain to be fully characterized, these data demonstrate an unexpected role for NF-kappaB induction during lytic reactivation in the production of replication-competent HHV-8 virions.
- Published
- 2004
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42. On the role of interferon regulatory factors in HIV-1 replication.
- Author
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Marsili G, Borsetti A, Sgarbanti M, Remoli AL, Ridolfi B, Stellacci E, Ensoli B, and Battistini A
- Subjects
- Humans, Models, Biological, Transcription Factors metabolism, Apoptosis physiology, HIV-1 physiology, Interferons physiology, Virus Replication physiology
- Abstract
Interferons (IFNs) are pleiotropic cytokines that possess several biological activities and play a central role in basic and applied research as mediators of antiviral and antigrowth responses, modulators of the immune system, and therapeutic agents against viral diseases and cancer. Interferon regulatory factors (IRFs) have been identified together with signal transducers and activators of transcription (STAT) from studies on the type I IFN as well as IFN-stimulated (ISG) gene regulation and signaling. IRFs constitute a family of transcriptional activators and repressors implicated in multiple biological processes including regulation of immune responses and host defence, cytokine signaling, cell growth regulation, and hematopoietic development. All members share a well-conserved DNA binding domain at the NH(2)-terminal region that recognizes similar DNA sequences, termed IRF element (IRF-E)/interferon-stimulated response element (ISRE), present on the promoter of target genes. Recently, a sequence homologous to the ISRE has been identified downstream from the 5' human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR). This sequence is a binding site for IRF-1 and IRF-2. Here we briefly summarize the role of IRFs in the regulation of HIV-1 LTR transcriptional activity and virus replication. The overall effect of IRFs on HIV-1 replication will also be discussed in the context of strategies carried out by the virus to counteract the IFN-mediated host defences both in active replication and during the establishment of viral latency.
- Published
- 2003
- Full Text
- View/download PDF
43. Disruption of the B-cell specific transcriptional program in HHV-8 associated primary effusion lymphoma cell lines.
- Author
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Arguello M, Sgarbanti M, Hernandez E, Mamane Y, Sharma S, Servant M, Lin R, and Hiscott J
- Subjects
- Body Fluids, Burkitt Lymphoma genetics, Burkitt Lymphoma metabolism, Burkitt Lymphoma pathology, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins genetics, Enhancer Elements, Genetic, Herpesviridae Infections metabolism, Herpesviridae Infections pathology, Hodgkin Disease genetics, Hodgkin Disease metabolism, Hodgkin Disease pathology, Humans, Immunoglobulin Light Chains genetics, Interferon Regulatory Factors, Lymphoma, B-Cell metabolism, Lymphoma, B-Cell pathology, Lymphoma, B-Cell virology, Neoplasm Proteins biosynthesis, Octamer Transcription Factor-2, PAX5 Transcription Factor, Proto-Oncogene Proteins biosynthesis, Proto-Oncogene Proteins genetics, RNA, Messenger biosynthesis, RNA, Neoplasm biosynthesis, Recombinant Fusion Proteins physiology, Repressor Proteins biosynthesis, Repressor Proteins genetics, Trans-Activators biosynthesis, Trans-Activators genetics, Transcription Factors biosynthesis, Transfection, Tumor Virus Infections metabolism, Tumor Virus Infections pathology, B-Lymphocytes metabolism, Gene Expression Regulation, Neoplastic, Herpesviridae Infections genetics, Herpesvirus 8, Human isolation & purification, Lymphoma, B-Cell genetics, Neoplasm Proteins genetics, Transcription Factors genetics, Transcription, Genetic, Tumor Virus Infections genetics
- Abstract
Primary effusion lymphoma (PEL) is a lymphoproliferative disease of B-cell origin that is associated with HHV-8 infection. PEL cells harbor a non-B, non-T phenotype and lack significant surface immunoglobulin (Ig) expression, a characteristic that has not been fully explained. In the present study, we demonstrate that PEL cells constitutively express interferon regulatory factor (IRF)-4, a transcription factor that regulates the activity of the immunoglobulin light-chain enhancer elements lambdaB and kappaE3' through binding to a composite Ets-IRF site. IRF-4 activity requires its physical interaction with PU.1, an Ets family member involved in the activation of genes essential for B-cell development. However, in PEL-derived B-cell lines, PU.1 expression was completely abrogated; expression of the B cell specific transcription factor Oct-2, which is known to regulate PU.1 expression, was also abolished. Moreover, the B-cell-specific coactivator of octamer factors, BOB-1/OcaB, was expressed at very decreased levels in PEL cells. Ectopic expression of Oct-2 was able to fully restore PU.1 promoter activity in the PEL cell line BCBL-1, while PU.1 expression also reconstituted the activity of the lambdaB Ets-IRF site. In addition, protein levels of BSAP/Pax-5 and IRF-8/ICSBP were undetectable in PEL cells. The pattern of transcription factor ablation observed in PEL was found to be comparable to that observed in classical Hodgkin's disease-derived cell lines, which also lack B-cell-specific surface markers. These observations indicate that disruption of the B-cell-specific transcriptional program is likely to contribute to the incomplete B-cell phenotype characteristic of PEL cells.
- Published
- 2003
- Full Text
- View/download PDF
44. Modulation of human immunodeficiency virus 1 replication by interferon regulatory factors.
- Author
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Sgarbanti M, Borsetti A, Moscufo N, Bellocchi MC, Ridolfi B, Nappi F, Marsili G, Marziali G, Coccia EM, Ensoli B, and Battistini A
- Subjects
- Cell Line, DNA-Binding Proteins genetics, Dose-Response Relationship, Drug, Electrophoretic Mobility Shift Assay, Gene Products, tat metabolism, HIV Long Terminal Repeat genetics, HIV-1 genetics, Humans, Interferon Regulatory Factor-1, Interferon Regulatory Factor-2, Interferon Regulatory Factors, Jurkat Cells, Phosphoproteins genetics, Protein Binding, RNA, Messenger genetics, RNA, Messenger metabolism, RNA, Viral genetics, RNA, Viral metabolism, Repressor Proteins genetics, Repressor Proteins metabolism, Response Elements genetics, Reverse Transcriptase Polymerase Chain Reaction, Transcription Factors genetics, Transcriptional Activation, tat Gene Products, Human Immunodeficiency Virus, DNA-Binding Proteins metabolism, Gene Expression Regulation, Viral, HIV-1 growth & development, Phosphoproteins metabolism, Transcription Factors metabolism, Virus Replication
- Abstract
Transcription of the human immunodeficiency virus (HIV)-1 is controlled by the cooperation of virally encoded and host regulatory proteins. The Tat protein is essential for viral replication, however, expression of Tat after virus entry requires HIV-1 promoter activation. A sequence in the 5' HIV-1 LTR, containing a binding site for transcription factors of the interferon regulatory factors (IRF) family has been suggested to be critical for HIV-1 transcription and replication. Here we show that IRF-1 activates HIV-1 LTR transcription in a dose-dependent fashion and in the absence of Tat. This has biological significance since IRF-1 is produced early upon virus entry, both in cell lines and in primary CD4+ T cells, and before expression of Tat. IRF-1 also cooperates with Tat in amplifying virus gene transcription and replication. This cooperation depends upon a physical interaction that is blocked by overexpression of IRF-8, the natural repressor of IRF-1, and, in turn is released by overexpression of IRF-1. These data suggest a key role of IRF-1 in the early phase of viral replication and/or during viral reactivation from latency, when viral transactivators are absent or present at very low levels, and suggest that the interplay between IRF-1 and IRF-8 may play a key role in virus latency.
- Published
- 2002
- Full Text
- View/download PDF
45. IRF regulation of HIV-1 long terminal repeat activity.
- Author
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Battistini A, Marsili G, Sgarbanti M, Ensoli B, and Hiscott J
- Subjects
- Base Sequence, Cyclic AMP Response Element-Binding Protein metabolism, Gene Expression Regulation, Viral, HIV-1 metabolism, Humans, Interferon Regulatory Factor-1, Interferon Regulatory Factor-2, Promoter Regions, Genetic, Response Elements, Sequence Homology, Transcription Factors physiology, Transcriptional Activation, DNA-Binding Proteins physiology, HIV Long Terminal Repeat, HIV-1 genetics, Phosphoproteins physiology, Repressor Proteins
- Abstract
Interferon (IFN) regulatory factors (IRF) constitute a family of transcriptional activators and repressors implicated in multiple biologic processes, including regulation of immune responses and host defense, cytokine signalling, cell growth regulation, and hematopoietic development. All members are characterized by well-conserved DNA binding domains at the N-terminal region that recognize similar DNA sequences termed IRF-binding element/IFN-stimulated response element (IRF-E/ISRE) present on the promoter of the IFN-alpha/beta genes and of some IFN-stimulated genes (ISG). Recently, a sequence homologous to the ISRE has been identified downstream of the 5' human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR). This sequence is a binding site for IRF-1 and IRF-2. Deletion of the LTR-ISRE results in impaired LTR promoter activity and decreased synthesis of viral RNA and proteins. Here, we briefly summarize characteristics of IRF-1 and IRF-2 binding to the HIV-1 LTR-ISRE and the data obtained to date on the functionality of this cis-element and on the role of IRF in the regulation of HIV-1 LTR transcriptional activity.
- Published
- 2002
- Full Text
- View/download PDF
46. HHV-8 encoded vIRF-1 represses the interferon antiviral response by blocking IRF-3 recruitment of the CBP/p300 coactivators.
- Author
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Lin R, Genin P, Mamane Y, Sgarbanti M, Battistini A, Harrington WJ Jr, Barber GN, and Hiscott J
- Subjects
- Antiviral Agents metabolism, Cells, Cultured, DNA-Binding Proteins genetics, Herpesvirus 8, Human genetics, Herpesvirus 8, Human pathogenicity, Humans, Interferon Regulatory Factor-3, Interferon Regulatory Factor-7, Interferon Regulatory Factors, Protein Binding, Transcription Factors genetics, Transcriptional Activation, Viral Proteins genetics, DNA-Binding Proteins metabolism, Herpesvirus 8, Human immunology, Interferons metabolism, Nuclear Proteins metabolism, Trans-Activators metabolism, Transcription Factors metabolism, Viral Proteins metabolism
- Abstract
Human herpes virus 8 (HHV-8) has developed unique mechanisms for altering cellular proliferative and apoptotic control pathways by incorporating viral homologs to several cellular regulatory genes into its genome. One of the important pirated genes encoded by the ORF K9 reading frame is a viral homolog of the interferon regulatory factors (IRF), a family of cellular transcription proteins that regulates expression of genes involved in pathogen response, immune modulation and cell proliferation. vIRF-1 has been shown to downregulate the interferon- and IRF-mediated transcriptional activation of ISG and murine IFNA4 gene promoters. In this study we demonstrate that vIRF-1 efficiently inhibited virus-induced expression of endogenous interferon B, CC chemokine RANTES and CXC chemokine IP-10 genes. Co-expression analysis revealed that vIRF-1 selectively blocked IRF-3 but not IRF-7-mediated transactivation. vIRF-1 was able to bind to both IRF-3 and IRF-7 in vivo as detected by coimmunoprecipitation analysis, but did not affect IRF-3 dimerization, nuclear translocation and DNA binding activity. Rather, vIRF-1 interacted with the CBP/p300 coactivators and efficiently inhibited the formation of transcriptionally competent IRF-3-CBP/p300 complexes. These results illustrate that vIRF-1 is able to block the early stages of the IFN response to virus infection by interfering with the activation of IRF-3 responsive, immediate early IFN genes.
- Published
- 2001
- Full Text
- View/download PDF
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