31 results on '"I. Pesce"'
Search Results
2. Reduced number and function of peripheral dendritic cells in coeliac disease
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B. Rovati, D. Piancatelli, G. Ricci, D. Millimaggi, S. Mazzocchi, M. Danova, Gino Roberto Corazza, Rachele Ciccocioppo, A. Cagnoni, and I. Pesce
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Adult ,Male ,medicine.medical_specialty ,Myeloid ,celiac ,Translational Studies ,Adolescent ,Lymphocyte ,Immunology ,Antigen presentation ,Cell Count ,Human leukocyte antigen ,Biology ,Gliadin ,Immunophenotyping ,Pathogenesis ,Aged ,Celiac Disease ,Cell Differentiation ,Cytokines ,Dendritic Cells ,Female ,Flow Cytometry ,Humans ,Lymphocyte Culture Test, Mixed ,Middle Aged ,Immune system ,Internal medicine ,medicine ,Immunology and Allergy ,Lymphocyte Culture Test ,Antigen-presenting cell ,Dendritic cell ,Mixed ,Endocrinology ,medicine.anatomical_structure - Abstract
Summary Dendritic cells (DC) play a pivotal role in shaping the immune response in both physiological and pathological conditions. In peripheral blood at least two subsets, the myeloid and plasmacytoid, have been described as having different T stimulatory functions and a variable degree of maturation. Certainly, antigen presentation plays a crucial role in the pathogenesis of coeliac disease and circulating immune cells are thought to reflect the state of immune response within the gut. Therefore, we aimed to investigate the quantitative and phenotypical modifications of peripheral blood DC, together with their functional properties, in this pathological condition. Blood samples from 11 untreated patients before and after a course of gluten-free diet, 27 treated patients and 14 controls underwent flow-cytometric analysis, while immunomagnetically sorted DC from the CD patients and eight human leucocyte antigen (HLA)-DQ2/8+ bone marrow donors were used to evaluate maturation status through the CD83 expression, cytokine profile for interleukin (IL)-6, IL-10, IL-12 and interferon (IFN)-α by enzyme-linked immunosorbent assay (ELISA), and functional properties by mixed leucocyte reaction before and after pulsing with digested gliadin. We found that in both untreated and treated patients, a significant reduction of the entire DC population, mainly the plasmacytoid subset, in comparison to healthy controls was observed. In active disease, an impaired allogenic lymphocyte reaction and a significant reduction of IFN-α production, paralleled by the presence of a more immature status, were also demonstrated. All the latter modifications have been reverted by pulsing DC with digested gliadin.
- Published
- 2007
3. 99. Auditory event-related potentials (ERPs) evoked by human syllables, musical notes/chords and animal sounds in pre-school children with specific expressive language disorders (SELDs) for assessing the selectiveness of auditory processing
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S. Alverà, D. Sarti, S. Magazù, I. Pesce, B. Molteni, S. Curzi, G. Airaghi, E. Granocchio, S. Giacomelli, V. Scaioli, and G. Musso
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Musical notation ,medicine.medical_specialty ,Rehabilitation ,Auditory event ,medicine.medical_treatment ,Sensory system ,Specific language impairment ,Audiology ,Stimulus (physiology) ,Neurophysiology ,medicine.disease ,behavioral disciplines and activities ,Sensory Systems ,Neurology ,Physiology (medical) ,medicine ,Pre school ,Neurology (clinical) ,Psychology - Abstract
Auditory ERP recordings are a useful neurophysiological means of assessing the temporal domain of the pre-linguistic and linguistic sensory deficits involved in expressive specific language impairment (SLI). We recorded auditory ERPs in 32 pre-school children aged 3–5 years with SLI before they underwent a rehabilitation programme. The ERPs were evoked using syllables (SyERPs), music (MuERPs) and animal sounds (AnERP) chosen from among those used in the trials carried out before, during and after an intensive rehabilitation programme in order assess: (a) the quality of ERP abnormalities; (b) the selectiveness of the ERP changes associated with the three types of stimuli; and (c) the changes induced by the rehabilitation programme. The SyERPs and MuERPs gave rise to four components (in P1, N1, N2 and P2) classified on the basis of their latency and polarity; the AnERPs only gave rise to P1 and N1 components. The ERPs had both common and distinctive features in terms of their waveforms and topographical distribution: the SyERPs were more left lateralised, and the MuERP more right lateralised. Later negative components were also recorded depending on the type of stimulus. In comparison with controls, P1 and P2 SyERPs were the most abnormal. As the rehabilitation led to selective SyERP changes, these may be useful for diagnosis and monitoring SLI.
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- 2016
4. Effects of gliadin stimulation on bone marrow-derived dendritic cells from HLA-DQ8 transgenic MICE
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Gino Roberto Corazza, Danilo Millimaggi, Mauro Rossi, Francesco Maurano, I. Pesce, G. Ricci, and Rachele Ciccocioppo
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CD4-Positive T-Lymphocytes ,Cells ,Receptor expression ,Bone Marrow Cells ,Mice, Transgenic ,Major histocompatibility complex ,Animals ,Celiac Disease ,Cell Differentiation ,Cells, Cultured ,Dendritic Cells ,Gliadin ,HLA-DQ Antigens ,Interferon-alpha ,Interleukins ,Lymphocyte Culture Test, Mixed ,Mice ,Mice, Inbred BALB C ,Toll-Like Receptors ,Transgenic ,Coeliac disease ,Flow cytometry ,medicine ,Lymphocyte Culture Test ,Inbred BALB C ,CD86 ,MHC class II ,Cultured ,Hepatology ,biology ,medicine.diagnostic_test ,Gastroenterology ,nutritional and metabolic diseases ,medicine.disease ,Molecular biology ,Mixed ,Immunology ,biology.protein ,CD80 - Abstract
BACKGROUND AND AIM: Gliadin presentation by HLA-DQ2/8 molecules to T cells plays a crucial role in triggering the inflammatory cascade in coeliac disease. We aimed to study the immunological effects of gliadin stimulation on dendritic cells (DCs) from HLA-DQ8 transgenic and BALB/c mice. METHODS: Bone marrow-derived DCs were stimulated with alpha-chymotrypsin-digested gliadin or ovoalbumin (100mug/ml). Modification of DC maturation, through HLA-DQ8 and MHC class II expression, and activation, by CD80 and CD86, was assessed by flow cytometry. The ability of pulsed and unpulsed DCs to prime T cells was evaluated by mixed leucocyte reaction. The expression of interleukin-4, -10, -12p70 and interferon-alpha, as well as of Toll-like receptor-4, -7, -8, -9 was determined by ELISA and real-time RT-PCR, respectively. RESULTS: Gliadin stimulation induced DC maturation (p
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- 2008
5. Correction of hypoxia and hypercapnia in COPD patients: effects on cerebrovascular flow
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G, Cannizzaro, L, Garbin, A, Clivati, and L I, Pesce
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Hypercapnia ,Male ,Ultrasonography, Doppler, Transcranial ,Cerebrovascular Circulation ,Oxygen Inhalation Therapy ,Humans ,Female ,Lung Diseases, Obstructive ,Middle Aged ,Hypoxia ,Blood Flow Velocity ,Intermittent Positive-Pressure Ventilation - Abstract
To assess the responsiveness of cerebral blood flow to arterial carbon dioxide tension (Pa,CO2), arterial oxygen tension (Pa,O2), and pH modifications, in chronic hypercapnia, we measured middle cerebral blood flow velocity (CBFV) by transcranial Doppler ultrasound in 13 chronically hypercapnic, long-term ventilated patients with chronic obstructive pulmonary disease (COPD), in the following conditions: 1) breathing room air; 2) with oxygen supplementation; 3) during mechanical noninvasive intermittent positive pressure ventilation (nIPPV) with O2 enrichment. Under baseline conditions (room air), the CBFV was within the normal range. During oxygen administration, a statistically significant increase was obtained in Pa,O2 (6.5 +/- 0.6 vs 11.2 +/- 1.9 kPa (49.1 +/- 4.3 vs 84.3 +/- 14.6 mmHg)), without relevant variations in: CBFV (54.2 +/- 9.1 cm.s-1), Pa,CO2 (8.6 +/- 1.0 kPa (64.7 +/- 7.7 mmHg)) and hydrogen ion concentration [H+] (42.9 +/- 2.9 nM), compared to baseline values (CBFV = 52.8 +/- 10.7 cm.s-1; Pa,CO2 = (8.4 +/- 0.9 kPa (63.1 +/- 7.1 mmHg; [H+] = 41.8 +/- 2.8 nM). After nIPPV, Pa,O2 did not increase any further (10.6 +/- 1.7 kPa (79.2 +/- 12.7 mmHg)), while CBFV (40.9 +/- 12.6 cm.s-1), Pa,CO2 (7.5 +/- 1.3 kPa (56.2 +/- 9.4 mmHg)) and [H+] (39.1 +/- 4.6 nM) showed a significant reduction compared to oxygen therapy (p0.01). We therefore conclude that in chronically hypercapnic long-term ventilated patients cerebral blood flow depends mainly on changes in Pa,CO2 and [H+], whilst oxygen does not seem to interfere with cerebral flow velocity. The reduction of Pa,CO2, due to mechanical ventilation, may determine cerebral blood vessel constriction, with possible impairment of cerebral functions.
- Published
- 1997
6. Clinical Activity and Safety of Anti-Programmed Death-1 (PD-1) (BMS-936558/MDX-1106/ONO-4538) in Patients (PTS) with Advanced Melanoma (MEL)
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Lada Mitchell, J. Pigozzo, K. Bennett, C.S.P. Lima, Silvia Park, Juan F. Medina, Antonio M. Grimaldi, David F. McDermott, Chi Hoon Maeng, R. Tanaka, L. Ridolfi, C.J.A. Punt, O.V. Korotkova, Amelia Lissia, D.M. Chen, W. Chang, J. De Vries, L. Pericleous, Ugo Marone, Corrado Caracò, Winald R. Gerritsen, E. Midena, C. Lebbé, Christian U. Blank, C. Brocia, E.F.D. Costa, R. Bassett, A. Sekulic, Tania Labiano, E. Fonsatti, D. Lawrence, Paola Queirolo, J.D. Wolchok, Stefano Mori, R. Dummer, C. Aliberti, Ruth Plummer, S. Francis, N. Vanhoutte, R. Wintherhalder, G.A.S. Nogueira, A. Amin, Jonathan D. Schwartz, M. Guida, C. Turtschi, Gerty Schreibelt, P. Mut, A. Ballesteros, S-H Lee, Anna C. Pavlick, Ester Simeone, M. Sini, K. Namikawa, R. Marconcini, Shibao Feng, Nicola Mozzillo, L. Veronese, C. Gamez, Merrick I. Ross, Donna Rowen, R. Labianca, T. Kirchhoff, M. Altomonte, Michele Maio, A. Batty, S. Yoo, James Larkin, Lev V. Demidov, Alessandro Testori, Omid Hamid, Sarvendra Kumar, Michael P. Brown, Jochen Utikal, J.A. Lopez Martin, R. Shapiro, Maria D. Lozano, N. Fischer, S. Ariad, B. Shafaeddin-Schreve, V. Chairion Sileni, M.K. Choi, Jung Yong Hong, Shreyaskumar Patel, Dimitris Bafaloukos, H. Yue, José I. Echeveste, M. Novy, M. Lebmeier, David R. Minor, F. Zambrana, M. Colombino, B. Campos, E. Muñoz, Simone M. Goldinger, D. Cumplido, P.L. Pilati, D. Lee, Giusy Gentilcore, G. Lucisano, J. Richards, Mario Sznol, F.S. Hodi, B. Merelli, Jeffrey S. Weber, M. Traversa, C. Oberkanins, Stephen M Murray, Suzanne L. Topalian, Vincent Brichard, I. Lazarev, D. Piazzalunga, F. De Galitiis, E. Wachter, C. Rubino, D. Opatt McDowell, Virginia Ferraresi, I.V. Samoylenko, M. Sereno, John A. Thompson, G. Colucci, P. Petrillo, M. Montañana, G. Di Monta, M. Maur, E. Bajetta, C. Oliveira, Kevin M. Chin, Sarah Danson, Anthony E. Oro, Igor Bondarenko, J.A. Rinck-Junior, W.J. Lesterhuis, E. Bertocci, A. Garcia Castano, T.N. Zabotina, S. Pisconti, S. Ellis, M. Hidalgo, A. Berrocal, Jeffrey A. Sosman, Sara Valpione, Miguel F. Sanmamed, Pier Francesco Ferrucci, Y. Sasajima, J. Perez, H. Linardou, F. De Rosa, J. Thompson, S. Stragliotto, Patrick Hwu, B.J. Coventry, M. Gillet, A.M. Di Giacomo, P.R. Hilfiker, L. Marchesi, Iman Osman, J. Rendleman, C. Nuzzo, G. Imberti, Edward McKenna, L. Di Guardo, Paul Nathan, I.N. Mikhaylova, Jenny Nobes, Antonio Cossu, Miguel Angel Idoate, Mario Mandalà, Giuseppe Palmieri, M. Ochoa de Olza, T. Nikoglou, M. Del Vecchio, B. Salaun, A. Cramarossa, J.M. Caminal, M. Biagioli, H. Tsuda, M.M. van Rossum, K. Harmankaya, J. Cortes, A.M. Moraes, H. Shaw, R. Danielli, S. Mosconi, John D. Hainsworth, Agop Y. Bedikian, G. Kriegshäuser, C.R. Scoggins, J. Valdivia, L. Pilla, R. Ridolfi, L.G. Campana, Christoph Rochlitz, M. Ma, V. Escrig, M.L. Cintra, I. Pesce, L. Calabrò, Karl D. Lewis, Russell S. Berman, Erik H.J.G. Aarntzen, Bart Neyns, T. Puertolas, J.A. Solomon, E. Castanon Alvarez, Georgia Kollia, F. Siannis, Katrin Conen, G.Z. Chkadua, Ana Arance, J.W. Lee, Caroline Robert, G.J. Lourenço, Jedd D. Wolchok, Lucia Benedetto, B.M. Smithers, N. Yamazaki, Axel Hauschild, A. Gupta, A. Gianatti, Luc Thomas, G. Rinaldi, A. Albano, D.P. Lawrence, F. Cognetti, A. Balogh, B. Rauscher, J.M. Wigginton, Carlo Tondini, W. Hwu, K. Baryshnikov, Y.S. Kim, A. Yakobson, J.M. Piulats, Ralf Gutzmer, Claus Garbe, R. Parrozzani, Kalijn F. Bol, M. Aglietta, V. Chiarion Sileni, Paolo A. Ascierto, M.R. Migden, P. Rojas, Nicholas E. Papadopoulos, V. De Giorgi, S. Martin Algarra, A. Tsutsumida, Ernie Marshall, S. Shang, S.V.L. Nicoletti, Joannes F M Jacobs, Anne Lynn S. Chang, J. Mayordomo, L. Cykowski, Sung Heon Kim, M. Gonzalez Cao, Sanjiv S. Agarwala, Michael S. Gordon, Carl G. Figdor, L. Alonso, Richard D. Carvajal, M.G. Bernengo, K. B. Kim, Daniela Massi, L. Dirix, O. Michielin, Nerea Gomez, Pippa Corrie, E. Ortega, Diana Giannarelli, E. Levchenko, H.R. Alexander, Alfonso Gurpide, P.M. LoRusso, Günther F.L. Hofbauer, J.D. Rinderknecht, B. Winn, L. Rivoltini, J. Hou, M. Aieta, S. Rossi, M.B. McHenry, Alejo Rodriguez-Vida, N. Eggmann, Alfred Zippelius, Y. Shao, G.J. Weiss, and Fabrizio Ayala
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Target lesion ,medicine.medical_specialty ,business.industry ,Melanoma ,Immediate family member ,Hematology ,medicine.disease ,Clinical trial ,Oncology ,Internal medicine ,Cohort ,medicine ,Previously treated ,business ,Survival rate ,Progressive disease - Abstract
Purpose BMS-936558 is a monoclonal antibody that blocks the PD-1 co-inhibitory receptor expressed by activated T cells. This study describes its activity and safety in pts with previously treated advanced MEL. Methods BMS-936558 was administered IV q2wk to pts with various tumors at 0.1 - 10 mg/kg during dose-escalation and/or cohort expansion. Pts received up to 12 cycles (4 doses/cycle) of treatment or until unacceptable toxicity, confirmed progressive disease, or complete response. Clinical activity was assessed by RECIST v1.0. Results As of Feb 24, 2012, 104 MEL pts had received BMS-936558 at 0.1 (n = 17), 0.3 (n = 19), 1 (n = 31), 3 (n = 17), or 10 mg/kg (n = 20). ECOG performance status was 0/1/2 in 63/38/3 pts, respectively. Most pts (67/104) had received prior immunotherapy (IT); prior anti-CTLA-4, -PD-1, or -PD-L1 was not permitted. The number of prior therapies was 1 (39%), 2 (35%), or ≥3 (26%). Median therapy duration was 20 wks (range 2.0 - 121.7 wks). The incidence of grade 3 - 4 related AEs was 20% and included gastrointestinal (4%), endocrine (2%), and hepatobiliary disorders (1%). There were no drug-related deaths in MEL pts. Clinical activity (responses or prolonged stable disease) was observed at all doses (Table). Of the 26/94 (28%) evaluable responders, 19 (73%) are ongoing ranging from 1.9+ to 24.9+ months. For the 23 responders followed ≥6 months from first dose on study, 16 (70%) are progression free. ORs occurred in pts with visceral or bone metastases. Six pts (6%; 95% CI 2 - 13%) had prolonged SD (≥24 wk); 3 pts had a persistent decrease in target lesion tumor burden in the presence of new lesions and were not categorized as responders. Conclusions BMS-936558 had durable clinical benefit in pts with advanced MEL, including those who had received prior IT. Additional long-term follow-up data will be reported. Dose, (mg/kg) No. ptsa ORR, No. pts (%) [95% CI] PFSR at 24 wk (%) [95% CI] 0.1 14 4 (29) [8 - 58] 40 [13 - 66] 0.3 16 3 (19) [4 - 46] 31 [9 - 54] 1 27 8 (30) [14 - 50] 45 [26 - 65] 3 17 7 (41) [18 - 67] * 55 [30 - 80] 10 20 4 (20) [6 - 44] 30 [9 - 51] * 1 CR aResponse-evaluable pts dosed by 7/01/2011 ORR = objective response rate ([{CR + PR} ÷ n] × 100); PFSR = progression-free survival rate. Disclosure J. Sosman: Research Funding: Bristol-Myers Squibb (myself). M. Sznol: Consultant or Advisory Role: Bristol-Myers Squibb (myself, compensated). Research Funding: Bristol-Myers Squibb (myself, clinical trials funding). D.F. McDermott: Advisory Board Role: Bristol-Myers Squibb (myself). R. Carvajal: Research Funding: Bristol-Myers Squibb (myself). S.L. Topalian: Consultant or Advisory Role: Bristol-Myers Squibb (myself, immediate family member, uncompensated). Research Funding: Bristol-Myers Squibb (myself). J.M. Wigginton: Employment or Leadership Position: Bristol-Myers Squibb (myself, employment, compensated). Stock Ownership: Bristol-Myers Squibb (myself). G. Kollia: Employment or Leadership Role: Bristol-Myers Squibb (employment, myself, compensated). Stock Ownership: Bristol-Myers Squibb/BMY stocks (myself). A. Gupta: Employment or Leadership Role: Bristol-Myers Squibb (employment, myself, compensated). Stock Ownership: Bristol-Myers Squibb (myself). F.S. Hodi: Consultant or Advisory Role: Bristol-Myers Squibb (myself, uncompensated). Research Funding: Bristol-Myers Squibb (myself). All other authors have declared no conflicts of interest.
- Published
- 2012
7. Clinical usefulness of a new portable oxygen concentrator
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L I, Pesce, G N, Bassi, and A, Santovito
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Adult ,Aged, 80 and over ,Male ,Oxygen Inhalation Therapy ,Humans ,Female ,Middle Aged ,Hypoxia ,Aged - Abstract
Thirty chronic, hypoxaemic patients with a mean arterial oxygen tension (PaO2) of 6.81 kPa (SD 0.56) in air were tested using the Travelair. The study was performed at rest, allowing the patients to breathe in the following conditions: a) compressed air; b) continuous oxygen flow from the concentrator; c) oxygen from the concentrator in demand-valve mode (DV) with an activation time (AT) of 375 ms; d) DV with AT of 750 ms; e) DV with AT of 1,125 ms; f) DV with AT 1,500 ms; and g) continuous oxygen from the hospital outlet at 2 l.min-1. The mean (SD) SaO2% values at each consecutive step were: a) 87.2 (5.0)%; b) 93.0 (3.0)%; c) 93.9 (2.8)%; d) 94.2 (2.5)%; e) 94.0 (2.7)%; f) 94.1 (2.7)%; and g) 94.8 (2.4)%, respectively. Each of the results obtained with DV (b-f) was statistically different from those obtained breathing air (a) or oxygen (g). The mean (SD) respiratory rates at each consecutive step were: a) 21.2 (4.1); b) 21.0 (4.0); c) 21.3 (4.0); d) 21.0 (3.6); e) 20.7 (3.4); f) 21.0 (3.8); and g) 21.2 (3.8) breaths.min-1, respectively. No relationship was found between the mean oxygen saturations and the mean respiratory frequencies of the patients in each condition tested. With the concentrator on demand, the average of the best SaO2% obtained by the patients in whatever of the four DV activation time modes (conditions c-f) was 94.8 (2.4)%, and no statistical difference was detected between this result and the SaO2% obtained with 2 l.min-1 of continuous oxygen.
- Published
- 1994
8. Wheat gliadin induces dendritic cell maturation and interferon-A production in HLA-DQ8 transgenic mice
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G.R. Corazza, G. Ricci, Rachele Ciccocioppo, E. Maurano, Danilo Millimaggi, F. Grandi, I. Pesce, and Mauro Rossi
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Genetically modified mouse ,Hepatology ,biology ,business.industry ,Interferon ,Gastroenterology ,biology.protein ,Medicine ,Dendritic cell ,Gliadin ,business ,Molecular biology ,medicine.drug - Published
- 2006
9. Characterization of peripheral blood dendritic cells in coeliac disease
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B. Rovati, G.R. Corazza, Danilo Millimaggi, Rachele Ciccocioppo, G. Ricci, A. Cagnoni, I. Pesce, M. Danova, and S. Mazzocchi
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Pathology ,medicine.medical_specialty ,Hepatology ,business.industry ,Gastroenterology ,Medicine ,business ,medicine.disease ,Coeliac disease ,Peripheral blood - Published
- 2006
10. LONG TERM SURVIVAL AND IMMUNOLOGICAL CORRELATES IN METASTATIC MELANOMA TREATED WITH IPILIMUMAB AT 10 MGS WITHIN AN EXPANDED ACCESS PROGRAM
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Ester Fonsatti, Maresa Altomonte, Luana Calabrò, Michele Maio, I. Pesce, Maurizio Biagioli, Riccardo Danielli, A.M. Di Giacomo, Diana Giannarelli, and Erica Bertocci
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medicine.medical_specialty ,Metastatic melanoma ,business.industry ,Ipilimumab ,Hematology ,Gastroenterology ,Peripheral blood mononuclear cell ,Oncology ,Internal medicine ,Expanded access ,Long term survival ,Cohort ,medicine ,Stage (cooking) ,business ,CD8 ,medicine.drug - Abstract
Background Ipilimumab (IPI) has shown long lasting responses in metastatic melanoma (MM) patients (pts) in phase II/III trials (Hodi et al., NEJM 2010; Robert et al. NEJM 2011; Prieto et al., CCR 2012). We have previously reported a significant clinical efficacy of IPI in 27 heavily pre-treated MM pts enrolled within the 10 mgs expanded access program (EAP) available at the University Hospital of Siena; one and 2-years survival were 34.8% and 23.5%, respectively (Di Giacomo, et al. CII, 2010). In spite of its clinical efficacy, no definitive predictive markers of response have been identified yet. We report here the survival follow-up of this cohort of MM pts and preliminary data on immunological correlates. Methods Twenty-seven pts with stage III (2) or IV (25) MM, progressing to a median of 3 (1-5) systemic therapies, were treated according to the EAP with IPI at 10 mg/Kg i.v. given at weeks (wks) 1, 4, 7, 10 during the induction phase, and every 12 wks from W24 in the maintenance phase. Peripheral blood mononuclear cells were collected from 17 pts at baseline, W7, W12 and W24 and analyzed by flow cytometry for ICOS expression on both CD4 and CD8+ T cells. The ratio between absolute blood neutrophils (N) and lymphocytes (L) count was determined at baseline, W4, W7 and W10 for 27, 27, 24 and 23 pts, respectively. Results With a median follow-up of 9.6 months the median OS was 9.6 months (95% CI: 4.2-16.1; range: 3.1-51.2; three and 4-years survival rates were 20.9%, with 5 long-term survivors (>4 yrs). A significant (p Conclusions ipi can induce long-term survivals in heavily pre-treated MM pts. Circulating ICOS+ T cells and N/L ratio in the course of treatment with ipi may represent predictive markers of clinical effectiveness in the daily practice. Disclosure M. Maio: Advisory Board and Honoraria from Bristol-Myers Squibb and Roche. All other authors have declared no conflicts of interest.
11. Guidelines for establishing a cytometry laboratory.
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Belkina AC, Roe CE, Tang VA, Back JB, Bispo C, Conway A, Chakraborty U, Daniels KT, de la Cruz G, Ferrer-Font L, Filby A, Gravano DM, Gregory MD, Hall C, Kukat C, Mozes A, Ordoñez-Rueda D, Orlowski-Oliver E, Pesce I, Porat Z, Poulton NJ, Reifel KM, Rieger AM, Sheridan RTC, Van Isterdael G, and Walker RV
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- Flow Cytometry, Laboratories, Software
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The purpose of this document is to provide guidance for establishing and maintaining growth and development of flow cytometry shared resource laboratories. While the best practices offered in this manuscript are not intended to be universal or exhaustive, they do outline key goals that should be prioritized to achieve operational excellence and meet the needs of the scientific community. Additionally, this document provides information on available technologies and software relevant to shared resource laboratories. This manuscript builds on the work of Barsky et al. 2016 published in Cytometry Part A and incorporates recent advancements in cytometric technology. A flow cytometer is a specialized piece of technology that require special care and consideration in its housing and operations. As with any scientific equipment, a thorough evaluation of the location, space requirements, auxiliary resources, and support is crucial for successful operation. This comprehensive resource has been written by past and present members of the International Society for Advancement of Cytometry (ISAC) Shared Resource Laboratory (SRL) Emerging Leaders Program https://isac-net.org/general/custom.asp?page=SRL-Emerging-Leaders with extensive expertise in managing flow cytometry SRLs from around the world in different settings including academia and industry. It is intended to assist in establishing a new flow cytometry SRL, re-purposing an existing space into such a facility, or adding a flow cytometer to an individual lab in academia or industry. This resource reviews the available cytometry technologies, the operational requirements, and best practices in SRL staffing and management., (© 2023 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)
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- 2024
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12. Integrating extracellular vesicle and circulating cell-free DNA analysis using a single plasma aliquot improves the detection of HER2 positivity in breast cancer patients.
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Mugoni V, Ciani Y, Quaini O, Tomasini S, Notarangelo M, Vannuccini F, Marinelli A, Leonardi E, Pontalti S, Martinelli A, Rossetto D, Pesce I, Mansy SS, Barbareschi M, Ferro A, Caffo O, Attard G, Di Vizio D, D'Agostino VG, Nardella C, and Demichelis F
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Multi-analyte liquid biopsies represent an emerging opportunity for non-invasive cancer assessment. We developed ONCE (One Aliquot for Circulating Elements), an approach for the isolation of extracellular vesicles (EV) and cell-free DNA (cfDNA) from a single aliquot of blood. We assessed ONCE performance to classify HER2-positive early-stage breast cancer (BrCa) patients by combining EV-associated RNA (EV-RNA) and cfDNA signals on n = 64 healthy donors (HD) and non-metastatic BrCa patients. Specifically, we isolated EV-enriched samples by a charge-based (CB) method and investigated EV-RNA and cfDNA by next-generation sequencing (NGS) and by digital droplet PCR (ddPCR). Sequencing of cfDNA and EV-RNA from HER2- and HER2+ patients demonstrated concordance with in situ molecular analyses of matched tissues. Combined analysis of the two circulating analytes by ddPCR showed increased sensitivity in ERBB2/HER2 detection compared to single nucleic acid components. Multi-analyte liquid biopsy prediction performance was comparable to tissue-based sequencing results from TCGA. Also, imaging flow cytometry analysis revealed HER2 protein on the surface of EV isolated from the HER2+ BrCa plasma, thus corroborating the potential relevance of studying EV as companion analyte to cfDNA. This data confirms the relevance of combining cfDNA and EV-RNA for HER2 cancer assessment and supports ONCE as a valuable tool for multi-analytes liquid biopsies' clinical implementation., Competing Interests: An Italian Patent Application no. 102021000027854 was filed on October 29th, 2021, on the ONCE methodology (authors involved are VM, YC, OQ, MN, VGD, CN, FD). All remaining authors have declared no conflicts of interest., (© 2023 The Authors. Journal of Extracellular Biology published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)
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- 2023
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13. Prediction of 2-[ 18 F]FDG PET-CT SUVmax for Adrenal Mass Characterization: A CT Radiomics Feasibility Study.
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Stanzione A, Cuocolo R, Bombace C, Pesce I, Mainolfi CG, De Giorgi M, Delli Paoli G, La Selva P, Petrone J, Camera L, Klain M, Del Vecchio S, Cuocolo A, and Maurea S
- Abstract
Background: Indeterminate adrenal masses (AM) pose a diagnostic challenge, and 2-[
18 F]FDG PET-CT serves as a problem-solving tool. Aim of this study was to investigate whether CT radiomics features could be used to predict the 2-[18 F]FDG SUVmax of AM., Methods: Patients with AM on 2-[18 F]FDG PET-CT scan were grouped based on iodine contrast injection as CT contrast-enhanced (CE) or CT unenhanced (NCE). Two-dimensional segmentations of AM were manually obtained by multiple operators on CT images. Image resampling and discretization (bin number = 16) were performed. 919 features were calculated using PyRadiomics. After scaling, unstable, redundant, and low variance features were discarded. Using linear regression and the Uniform Manifold Approximation and Projection technique, a CT radiomics synthetic value (RadSV) was obtained. The correlation between CT RadSV and 2-[18 F]FDG SUVmax was assessed with Pearson test., Results: A total of 725 patients underwent PET-CT from April 2020 to April 2021. In 150 (21%) patients, a total of 179 AM (29 bilateral) were detected. Group CE consisted of 84 patients with 108 AM (size = 18.1 ± 4.9 mm) and Group NCE of 66 patients with 71 AM (size = 18.5 ± 3.8 mm). In both groups, 39 features were selected. No statisticallyf significant correlation between CT RadSV and 2-[18 F]FDG SUVmax was found (Group CE, r = 0.18 and p = 0.058; Group NCE, r = 0.13 and p = 0.27)., Conclusions: It might not be feasible to predict 2-[18 F]FDG SUVmax of AM using CT RadSV. Its role as a problem-solving tool for indeterminate AM remains fundamental.- Published
- 2023
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14. The Role of Physical Exercise and Rehabilitative Implications in the Process of Nerve Repair in Peripheral Neuropathies: A Systematic Review.
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Chiaramonte R, Pavone V, Testa G, Pesce I, Scaturro D, Musumeci G, Mauro GL, and Vecchio M
- Abstract
Background: The various mechanisms involved in peripheral nerve regeneration, induced by exercise and electrical nerve stimulation, are still unclear., Objective: The aim of this review was to summarize the influence of physical exercise and/or electrical stimulation on peripheral nerve repair and regeneration and the variation of impact of intervention depending on timing, as well as kind and dosage of the intervention. A literature survey was conducted on PubMed, Scopus, and Web of Science, between February 2021 to July 2021, with an update in September 2022., Methodology: The literature search identified 101,386 articles with the keywords: "peripheral nerve" OR "neuropathy" AND "sprouting" OR "neuroapraxia" OR "axonotmesis" OR "neurotmesis" OR "muscle denervation" OR "denervated muscle" AND "rehabilitation" OR "physical activity" OR "physical exercise" OR "activity" OR "electrical stimulation". A total of 60 publications were included. Eligible studies were focused on evaluating the process of nerve repair (biopsy, electromyographic parameters or biomarker outcomes) after electrical stimulation or physical exercise interventions on humans or animals with peripheral sensory or motor nerve injury., Synthesis: This study shows that the literature, especially regarding preclinical research, is mainly in agreement that an early physical program with active exercise and/or electrical stimulation promotes axonal regenerative responses and prevents maladaptive response. This was evaluated by means of changes in electrophysiological recordings of CMAPs for latency amplitude, and the sciatic functional index (SFI). Furthermore, this type of activity can cause an increase in weight and in muscle fiber diameter. Nevertheless, some detrimental effects of exercising and electrical stimulation too early after nerve repair were recorded., Conclusion: In most preclinical studies, peripheral neuropathy function was associated with improvements after physical exercise and electrical stimulation. For humans, too little research has been conducted on this topic to reach a complete conclusion. This research supports the need for future studies to test the validity of a possible rehabilitation treatment in humans in cases of peripheral neuropathy to help nerve sprouting.
- Published
- 2023
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15. Translational enhancement by base editing of the Kozak sequence rescues haploinsufficiency.
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Ambrosini C, Destefanis E, Kheir E, Broso F, Alessandrini F, Longhi S, Battisti N, Pesce I, Dassi E, Petris G, Cereseto A, and Quattrone A
- Subjects
- Alleles, Codon, Initiator, Humans, RNA, Messenger metabolism, Haploinsufficiency genetics, Nucleotides
- Abstract
A variety of single-gene human diseases are caused by haploinsufficiency, a genetic condition by which mutational inactivation of one allele leads to reduced protein levels and functional impairment. Translational enhancement of the spare allele could exert a therapeutic effect. Here we developed BOOST, a novel gene-editing approach to rescue haploinsufficiency loci by the change of specific single nucleotides in the Kozak sequence, which controls translation by regulating start codon recognition. We evaluated for translational strength 230 Kozak sequences of annotated human haploinsufficient genes and 4621 derived variants, which can be installed by base editing, by a high-throughput reporter assay. Of these variants, 149 increased the translation of 47 Kozak sequences, demonstrating that a substantial proportion of haploinsufficient genes are controlled by suboptimal Kozak sequences. Validation of 18 variants for 8 genes produced an average enhancement in an expression window compatible with the rescue of the genetic imbalance. Base editing of the NCF1 gene, whose monoallelic loss causes chronic granulomatous disease, resulted in the desired increase of NCF1 (p47phox) protein levels in a relevant cell model. We propose BOOST as a fine-tuned approach to modulate translation, applicable to the correction of dozens of haploinsufficient monogenic disorders independently of the causing mutation., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2022
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16. Zebrafish Melanoma-Derived Interstitial EVs Are Carriers of ncRNAs That Induce Inflammation.
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Biagini V, Busi F, Anelli V, Kerschbamer E, Baghini M, Gurrieri E, Notarangelo M, Pesce I, van Niel G, D'Agostino VG, and Mione M
- Subjects
- Animals, Inflammation genetics, Inflammation metabolism, RNA, Untranslated metabolism, Zebrafish genetics, Extracellular Vesicles metabolism, Melanoma genetics, Melanoma metabolism
- Abstract
Extracellular vesicles (EVs) are membranous particles released by all cell types. Their role as functional carrier of bioactive molecules is boosted by cells that actively secrete them in biological fluids or in the intercellular space (interstitial EVs, iEVs). Here we have optimised a method for the isolation and characterization of zebrafish iEVs from whole melanoma tissues. Zebrafish melanoma iEVs are around 140 nm in diameter, as determined by nanoparticle tracking and transmission electron microscopy (TEM) analysis. Western blot analysis shows enrichment for CD63 and Alix in the iEV fraction, but not in melanoma cell lysates. Super resolution and confocal microscopy reveal that purified zebrafish iEVs are green fluorescent protein positive (GFP+), indicating that they integrate the oncogene GFP-HRAS
V12G used to induce melanoma in this model within their vesicular membrane or luminal content. Analysis of RNA-Seq data found 118 non-coding (nc)RNAs differentially distributed between zebrafish melanoma and their iEVs, with only 17 of them being selectively enriched in iEVs. Among these, the RNA components of RNAses P and MRP, which process ribosomal RNA precursors, mitochondrial RNAs, and some mRNAs, were enriched in zebrafish and human melanoma EVs, but not in iEVs extracted from brain tumours. We found that melanoma iEVs induce an inflammatory response when injected in larvae, with increased expression of interferon responsive genes, and this effect is reproduced by MRP- or P-RNAs injected into circulation. This suggests that zebrafish melanoma iEVs are a source of MRP- and P-RNAs that can trigger inflammation in cells of the innate immune system.- Published
- 2022
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17. Metastatic choriocarcinoma with hemorrhagic complications and spontaneous ovarian hyperstimulation syndrome: A case report.
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De Lucia DR, Castaldo A, D'Agostino V, Ascione R, Pesce I, Coppola L, Catelli A, and Radice L
- Abstract
Gestational choriocarcinoma is a malignant trophoblastic tumor arising from any gestational event, even with a long latency period, generally in the reproductive female. It is associated with a high level of beta-human chorionic gonadotropin. Its primary site is usually the uterus but not all patients have a detectable lesion in this site. Regression of the primary tumor after it has metastasized is not uncommon, and one-third of cases manifest as complications of metastatic disease. In this report we present an uncommon case of gestational choriocarcinoma with lung, liver and jejunal metastases at the time of diagnosis without evidence of pelvic disease, in 34-year-old woman. The main points of interest of our case were the development of the ovarian hyperstimulation syndrome with massive multicystic ovarian enlargement induced by high level of beta-human chorionic gonadotropin and the bleeding of jejunal and liver metastases, due to the high vascularity of the tumor tissue, a condition known as "Choriocarcinoma Syndrome". We will focus on the radiological findings of metastases, bleeding complications and ovarian hyperstimulation syndrome., (© 2021 The Authors. Published by Elsevier Inc. on behalf of University of Washington.)
- Published
- 2021
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18. The role of unhenanced CT in a ruptured parathyroid adenoma: A case report.
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Lombardi R, Muccio CF, D'Auria V, D'Agostino V, Genovese S, Monaci A, La Selva P, Castaldo A, Pesce I, and Bencivenga A
- Abstract
Acute bleeding is a rare and potentially life-threatening complication of a Parathyroid Adenoma described in just a few cases in literature. We describe the case of a healthy 53-years-old female patient without prior history of parathyroid pathology who presented with acute onset of neck and mediastinal hemorrhage. Ultrasound (US), Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) combined with laboratory tests led to the diagnosis of a bleeding Parathyroid adenoma. This case is presented to sensitize both Radiologists and Clinicians about this rare presentation that should be put into differential diagnosis of acute neck swelling and pain., (© 2021 The Authors. Published by Elsevier Inc. on behalf of University of Washington.)
- Published
- 2021
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19. An ileal duplication cyst case report: From diagnosis to treatment.
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D'Agostino V, Castaldo A, Catelli A, Pesce I, Genovese S, Coppola L, Monaci A, Esposito C, and Amitrano M
- Abstract
Enteric duplication cysts (EDCs) are rare congenital malformations of the children and can develop everywhere along the gastrointestinal (GI) tract, being the ileum the most frequent localization. We herein present an unusual case of duplication cyst of ileal origin who show a tubular morphology and doesn't communicate with GI lumen. A 2-month-old boy was admitted to our hospital for investigation of an anechoic formation of the lower right abdomen for the surgical planning. The patient was asymptomatic. Ultrasound (US) and magnetic resonance imaging (MRI) showed features of a cystic lesion. Laparoscopic surgery was performed and the cyst excised. Macroscopic examination and histologic findings confirmed the diagnosis of a enteric duplication cyst arising from the ileum. In a patient with an abdominal cystic mass, although asymptomatic, it's worth assessing the nature of the lesion and planning a surgery in order to avoid future complications. A correct use of diagnostic it's fundamental to identify the etiology and the characteristics of a cystic mass., (© 2021 The Authors. Published by Elsevier Inc. on behalf of University of Washington.)
- Published
- 2021
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20. Solutions for Shared Resource Lab Remote Quality Control and Instrument Troubleshooting during a Pandemic.
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Gravano DM, Chakraborty U, Pesce I, and Thomson M
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- COVID-19 prevention & control, Flow Cytometry trends, Humans, Laboratories trends, Teleworking trends, Webcasts as Topic standards, Webcasts as Topic trends, Workflow, COVID-19 epidemiology, Flow Cytometry instrumentation, Flow Cytometry standards, Laboratories standards, Quality Control, Teleworking standards
- Abstract
The COVID-19 pandemic has dramatically affected shared resource lab (SRL) staff in-person availability at institutions globally. This article discusses the challenges of ensuring reliable instrument performance and quality data output while facility staff and external service provider on-site presence is severely limited. Solutions revolve around the adoption of remote monitoring and troubleshooting platforms, provision of self-service troubleshooting resources specific to facility instruments and workflows, development of an assistance contact policy, and ensuring efficiency of limited in-person staff time. These solutions employ software and hardware tools that are already in use or readily available in the SRL community, such as remote instrument access tools, video hosting and conferencing platforms, and ISAC shared resources. © 2020 International Society for Advancement of Cytometry., (© 2020 International Society for Advancement of Cytometry.)
- Published
- 2021
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21. Regional anesthesia during the COVID-19 pandemic: a time to reconsider practices? (Letter #2).
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Aliste J, Altermatt FR, Atton R, Bravo D, Layera S, Miranda P, and Pesce I
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- COVID-19, Humans, SARS-CoV-2, Anesthesia, Conduction, Betacoronavirus, Coronavirus Infections, Pandemics, Pneumonia, Viral
- Published
- 2020
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22. Application of CRISPR/Cas9 editing and digital droplet PCR in human iPSCs to generate novel knock-in reporter lines to visualize dopaminergic neurons.
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Überbacher C, Obergasteiger J, Volta M, Venezia S, Müller S, Pesce I, Pizzi S, Lamonaca G, Picard A, Cattelan G, Malpeli G, Zoli M, Beccano-Kelly D, Flynn R, Wade-Martins R, Pramstaller PP, Hicks AA, Cowley SA, and Corti C
- Subjects
- Cell Line, Dopaminergic Neurons cytology, Green Fluorescent Proteins genetics, Humans, Induced Pluripotent Stem Cells cytology, Microscopy, Fluorescence, CRISPR-Cas Systems, Dopaminergic Neurons metabolism, Gene Editing, Gene Knock-In Techniques, Green Fluorescent Proteins biosynthesis, Induced Pluripotent Stem Cells metabolism, Polymerase Chain Reaction, Transgenes
- Abstract
Human induced pluripotent stem cells (hiPSCs) have become indispensable for disease modelling. They are an important resource to access patient cells harbouring disease-causing mutations. Derivation of midbrain dopaminergic (DAergic) neurons from hiPSCs of PD patients represents the only option to model physiological processes in a cell type that is not otherwise accessible from human patients. However, differentiation does not produce a homogenous population of DA neurons and contaminant cell types may interfere with the readout of the in vitro system. Here, we use CRISPR/Cas9 to generate novel knock-in reporter lines for DA neurons, engineered with an endogenous fluorescent tyrosine hydroxylase - enhanced green fluorescent protein (TH-eGFP) reporter. We present a reproducible knock-in strategy combined with a highly specific homologous directed repair (HDR) screening approach using digital droplet PCR (ddPCR). The knock-in cell lines that we created show a functioning fluorescent reporter system for DA neurons that are identifiable by flow cytometry., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2019
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23. Ultrasensitive detection of cancer biomarkers by nickel-based isolation of polydisperse extracellular vesicles from blood.
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Notarangelo M, Zucal C, Modelska A, Pesce I, Scarduelli G, Potrich C, Lunelli L, Pederzolli C, Pavan P, la Marca G, Pasini L, Ulivi P, Beltran H, Demichelis F, Provenzani A, Quattrone A, and D'Agostino VG
- Subjects
- Case-Control Studies, Cell Line, Tumor, Flow Cytometry, Humans, Liquid Biopsy standards, Neoplasms genetics, Neoplasms metabolism, Polymerase Chain Reaction, Sensitivity and Specificity, Ultracentrifugation, Biomarkers, Tumor blood, Extracellular Vesicles metabolism, Extracellular Vesicles ultrastructure, Liquid Biopsy methods, Neoplasms blood, Neoplasms diagnosis, Nickel
- Abstract
Background: Extracellular vesicles (EVs) are secreted membranous particles intensively studied for their potential cargo of diagnostic markers. Efficient and cost-effective isolation methods need to be established for the reproducible and high-throughput study of EVs in the clinical practice., Methods: We designed the nickel-based isolation (NBI) to rapidly isolate EVs and combined it with newly-designed amplified luminescent proximity homogeneous assay or digital PCR to detect biomarkers of clinical utility., Findings: From plasma of 46 healthy donors, we systematically recovered small EV (~250 nm of mean diameter; ~3 × 10
10 /ml) and large EV (~560 nm of mean diameter; ~5 × 108 /ml) lineages ranging from 50 to 700 nm, which displayed hematopoietic/endothelial cell markers that were also used in spike-in experiments using EVs from tumor cell lines. In retrospective studies, we detected picomolar concentrations of prostate-specific membrane antigen (PSMA) in fractions of EVs isolated from the plasma of prostate cancer patients, discriminating them from control subjects. Directly from oil-encapsulated EVs for digital PCR, we identified somatic BRAF and KRAS mutations circulating in the plasma of metastatic colorectal cancer (CRC) patients, matching 100% of concordance with tissue diagnostics. Importantly, with higher sensitivity and specificity compared with immuno-isolated EVs, we revealed additional somatic alterations in 7% of wild-type CRC cases that were subsequently validated by further inspections in the matched tissue biopsies., Interpretation: We propose NBI-combined approaches as simple, fast, and robust strategies to probe the tumor heterogeneity and contribute to the development of EV-based liquid biopsy studies. FUND: Associazione Italiana per la Ricerca sul Cancro (AIRC), Fondazione Cassa di Risparmio Trento e Rovereto (CARITRO), and the Italian Ministero Istruzione, Università e Ricerca (Miur)., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2019
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24. Modulation of endotoxicity of Shigella generalized modules for membrane antigens (GMMA) by genetic lipid A modifications: relative activation of TLR4 and TLR2 pathways in different mutants.
- Author
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Rossi O, Pesce I, Giannelli C, Aprea S, Caboni M, Citiulo F, Valentini S, Ferlenghi I, MacLennan CA, D'Oro U, Saul A, and Gerke C
- Subjects
- Acylation immunology, Acyltransferases genetics, Acyltransferases metabolism, Antigens, Bacterial genetics, Antigens, Bacterial metabolism, Bacterial Outer Membrane Proteins genetics, Bacterial Outer Membrane Proteins metabolism, Bacterial Proteins genetics, Bacterial Proteins metabolism, Cells, Cultured, Cytokines immunology, Cytokines metabolism, Electrophoresis, Polyacrylamide Gel, HEK293 Cells, Humans, Lipid A analysis, Lipid A metabolism, Microscopy, Electron, Transmission, Monocytes immunology, Monocytes metabolism, Mutation, Shigella genetics, Shigella metabolism, Shigella flexneri genetics, Shigella flexneri immunology, Shigella flexneri metabolism, Shigella sonnei genetics, Shigella sonnei immunology, Shigella sonnei metabolism, Signal Transduction immunology, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Toll-Like Receptor 2 metabolism, Toll-Like Receptor 4 metabolism, Antigens, Bacterial immunology, Bacterial Outer Membrane Proteins immunology, Lipid A immunology, Shigella immunology, Toll-Like Receptor 2 immunology, Toll-Like Receptor 4 immunology
- Abstract
Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from Shigella. As GMMA are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (LPS). We examined ways of reducing their reactogenicity by modifying lipid A, the endotoxic part of LPS, through deletion of late acyltransferase genes, msbB or htrB, in GMMA-producing Shigella sonnei and Shigella flexneri strains. GMMA with resulting penta-acylated lipid A from the msbB mutants showed a 600-fold reduced ability, and GMMA from the S. sonnei ΔhtrB mutant showed a 60,000-fold reduced ability compared with GMMA with wild-type lipid A to stimulate human Toll-like receptor 4 (TLR4) in a reporter cell line. In human peripheral blood mononuclear cells, GMMA with penta-acylated lipid A showed a marked reduction in induction of inflammatory cytokines (S. sonnei ΔhtrB, 800-fold; ΔmsbB mutants, 300-fold). We found that the residual activity of these GMMA is largely due to non-lipid A-related TLR2 activation. In contrast, in the S. flexneri ΔhtrB mutant, a compensatory lipid A palmitoleoylation resulted in GMMA with hexa-acylated lipid A with ∼10-fold higher activity to stimulate peripheral blood mononuclear cells than GMMA with penta-acylated lipid A, mostly due to retained TLR4 activity. Thus, for use as vaccines, GMMA will likely require lipid A penta-acylation. The results identify the relative contributions of TLR4 and TLR2 activation by GMMA, which need to be taken into consideration for GMMA vaccine development., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2014
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25. Long-term survival and immunological parameters in metastatic melanoma patients who responded to ipilimumab 10 mg/kg within an expanded access programme.
- Author
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Di Giacomo AM, Calabrò L, Danielli R, Fonsatti E, Bertocci E, Pesce I, Fazio C, Cutaia O, Giannarelli D, Miracco C, Biagioli M, Altomonte M, and Maio M
- Subjects
- Adult, Aged, Antibodies, Monoclonal adverse effects, Antineoplastic Agents adverse effects, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, Female, Humans, Inducible T-Cell Co-Stimulator Protein metabolism, Ipilimumab, Lymphocyte Count, Male, Melanoma immunology, Melanoma pathology, Middle Aged, Neoplasm Metastasis, Neoplasm Staging, Retrospective Studies, Treatment Outcome, Young Adult, Antibodies, Monoclonal administration & dosage, Antineoplastic Agents administration & dosage, Melanoma drug therapy, Melanoma mortality
- Abstract
Background: Ipilimumab can result in durable clinical responses among patients with advanced melanoma. However, no predictive marker of clinical activity has yet been identified. We provide preliminary data describing the correlation between immunological parameters and response/survival among patients with advanced melanoma who received ipilimumab 10 mg/kg in an expanded access programme., Methods: Patients received ipilimumab 10 mg/kg every 3 weeks (Q3W) for four doses (induction) and Q12W from week 24 (W24) as maintenance therapy. Tumor assessments were conducted Q12W. Expression of inducible T cell costimulator (ICOS) on CD4(+) and CD8(+) T cells was assessed at baseline, W7, W12 and W24, and the ratio between absolute neutrophils (N) and lymphocytes (L) determined at baseline, W4, W7 and W10., Results: Median overall survival among 27 patients was 9.6 months (95 % CI 3.2-16.1), with 3- and 4-year survival rates of 20.4 %. Five patients survived >4 years. Patients with an increase in the number of circulating ICOS(+) T cells at W7 were more likely to experience disease control and have improved survival. An N/L ratio below the median at W7 and W10 was also associated with better survival compared with an N/L ratio above the median., Conclusions: Ipilimumab can induce long-term survival benefits in heavily pretreated patients with metastatic melanoma. Changes in the number of circulating ICOS(+) T cells or N/L ratio during ipilimumab treatment may represent early markers of response. However, given the limited sample size, further investigation is required.
- Published
- 2013
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26. High yield production process for Shigella outer membrane particles.
- Author
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Berlanda Scorza F, Colucci AM, Maggiore L, Sanzone S, Rossi O, Ferlenghi I, Pesce I, Caboni M, Norais N, Di Cioccio V, Saul A, and Gerke C
- Subjects
- Animals, Antigens, Surface isolation & purification, Blotting, Western, Computational Biology, DNA Primers genetics, Electrophoresis, Gel, Two-Dimensional, Enzyme-Linked Immunosorbent Assay, Female, Fermentation, Gene Knockout Techniques, Mice, Microscopy, Electron, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Vaccines biosynthesis, Bacterial Outer Membrane Proteins metabolism, Biotechnology methods, Membrane Lipids metabolism, Protein Engineering methods, Shigella sonnei metabolism
- Abstract
Gram-negative bacteria naturally shed particles that consist of outer membrane lipids, outer membrane proteins, and soluble periplasmic components. These particles have been proposed for use as vaccines but the yield has been problematic. We developed a high yielding production process of genetically derived outer membrane particles from the human pathogen Shigella sonnei. Yields of approximately 100 milligrams of membrane-associated proteins per liter of fermentation were obtained from cultures of S. sonnei ΔtolR ΔgalU at optical densities of 30-45 in a 5 L fermenter. Proteomic analysis of the purified particles showed the preparation to primarily contain predicted outer membrane and periplasmic proteins. These were highly immunogenic in mice. The production of these outer membrane particles from high density cultivation of bacteria supports the feasibility of scaling up this approach as an affordable manufacturing process. Furthermore, we demonstrate the feasibility of using this process with other genetic manipulations e.g. abolition of O antigen synthesis and modification of the lipopolysaccharide structure in order to modify the immunogenicity or reactogenicity of the particles. This work provides the basis for a large scale manufacturing process of Generalized Modules of Membrane Antigens (GMMA) for production of vaccines from gram-negative bacteria.
- Published
- 2012
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27. Intranasal administration of CpG induces a rapid and transient cytokine response followed by dendritic and natural killer cell activation and recruitment in the mouse lung.
- Author
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Pesce I, Monaci E, Muzzi A, Tritto E, Tavarini S, Nuti S, De Gregorio E, and Wack A
- Subjects
- Administration, Intranasal, Animals, B-Lymphocytes immunology, Bronchoalveolar Lavage Fluid cytology, Bronchoalveolar Lavage Fluid immunology, CD11c Antigen metabolism, Female, Lung cytology, Lymphocyte Activation immunology, Macrophages, Alveolar immunology, Mice, Mice, Inbred BALB C, Oligodeoxyribonucleotides immunology, Adjuvants, Immunologic administration & dosage, Cytokines metabolism, Dendritic Cells immunology, Killer Cells, Natural immunology, Lung immunology, Oligodeoxyribonucleotides administration & dosage
- Abstract
CpG-containing oligodeoxynucleotides are potent mucosal adjuvants and effective as stand-alone treatment of respiratory infections in mice. Although CpG is also used as a type 1 helper immunomodulator in the treatment of asthma and allergic disease, immune modulation following intranasal application has not been fully characterized yet. Using a B-type CpG, we monitored RNA expression profiles, cytokine production and cellular activation in lung tissue and bronchoalveolar lavages ex vivo and cytokine production of purified cell populations in vitro. CpG triggered the upregulation of many transcripts, including interferon response genes and proinflammatory cytokine genes, between 3 h and 4 days. Overlapping subsets of these cytokine proteins were induced in vitro in purified CD11c+ cells, B cells and alveolar macrophages from the lung, thus identifying these cells as direct targets of CpG. While lung B cells strongly respond to CpG in vitro, less activation is found ex vivo, suggesting efficient CpG sequestering or rapid B cell migration after activation. In contrast, a type II alveolar epithelial cell line did not respond to CpG in vitro. We noted selective recruitment of plasmacytoid dendritic cells (DCs) into the lung tissue, and of conventional DCs and natural killer (NK) cells into the lung tissue and bronchoalveolar space. Furthermore, CpG induced activation of intrapulmonary DCs, NK and T cells. We hypothesize that CpG-linked adjuvanticity and clearance of respiratory pathogens are mediated by two major mechanisms: transient induction of the interferon pathway limiting microbial survival and selective recruitment of DCs and NK cells, which allows for better adaptive responses.
- Published
- 2010
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28. Effects of gliadin stimulation on bone marrow-derived dendritic cells from HLA-DQ8 transgenic MICE.
- Author
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Ciccocioppo R, Rossi M, Pesce I, Ricci G, Millimaggi D, Maurano F, and Corazza GR
- Subjects
- Animals, CD4-Positive T-Lymphocytes physiology, Cells, Cultured, Dendritic Cells metabolism, HLA-DQ Antigens, Interferon-alpha metabolism, Interleukins metabolism, Lymphocyte Culture Test, Mixed, Mice, Mice, Inbred BALB C, Mice, Transgenic, Toll-Like Receptors metabolism, Bone Marrow Cells physiology, Celiac Disease physiopathology, Cell Differentiation drug effects, Dendritic Cells drug effects, Gliadin pharmacology
- Abstract
Background and Aim: Gliadin presentation by HLA-DQ2/8 molecules to T cells plays a crucial role in triggering the inflammatory cascade in coeliac disease. We aimed to study the immunological effects of gliadin stimulation on dendritic cells (DCs) from HLA-DQ8 transgenic and BALB/c mice., Methods: Bone marrow-derived DCs were stimulated with alpha-chymotrypsin-digested gliadin or ovoalbumin (100 microg/ml). Modification of DC maturation, through HLA-DQ8 and MHC class II expression, and activation, by CD80 and CD86, was assessed by flow cytometry. The ability of pulsed and unpulsed DCs to prime T cells was evaluated by mixed leucocyte reaction. The expression of interleukin-4, -10, -12p70 and interferon-alpha, as well as of Toll-like receptor-4, -7, -8, -9 was determined by ELISA and real-time RT-PCR, respectively., Results: Gliadin stimulation induced DC maturation (p<0.001 in BALB/c, p<0.01 in DQ8) but not activation, whereas ovoalbumin upregulated all markers (p<0.01 for maturation and p<0.001 for activation in both DC populations). No increase of T proliferation was elicited by pulsed DCs with respect to unpulsed DCs. Only in DQ8 DCs, gliadin induced Toll-like receptor-4 (p<0.001), -7 (p<0.001), -8 (p<0.005) expression and interferon-alpha (p<0.001) secretion., Conclusion: Gliadin resulted unable to activate DC, but stimulated Toll-like receptor expression and interferon-alpha secretion.
- Published
- 2008
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29. The acquired immune response to the mucosal adjuvant LTK63 imprints the mouse lung with a protective signature.
- Author
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Tritto E, Muzzi A, Pesce I, Monaci E, Nuti S, Galli G, Wack A, Rappuoli R, Hussell T, and De Gregorio E
- Subjects
- Animals, B-Lymphocyte Subsets immunology, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, Cell Movement immunology, Chemokines biosynthesis, Chemokines genetics, Flow Cytometry, Gene Expression Profiling, Immunity, Cellular, Immunity, Innate, Intubation, Intratracheal, Lung cytology, Lymphocyte Activation immunology, Mice, Mice, Inbred BALB C, Mice, Nude, Mice, SCID, Transcription, Genetic immunology, Adjuvants, Immunologic administration & dosage, Bacterial Toxins administration & dosage, Bacterial Toxins immunology, Enterotoxins administration & dosage, Enterotoxins immunology, Escherichia coli Proteins administration & dosage, Escherichia coli Proteins immunology, Immunity, Mucosal, Lung immunology, Lung metabolism
- Abstract
LTK63, a nontoxic mutant of Escherichia coli heat labile enterotoxin (LT), is a potent and safe mucosal adjuvant that has also been shown to confer generic protection to several respiratory pathogens. To understand the mechanisms of action underlying the LTK63 protective effect, we analyzed the molecular and cellular events triggered by its administration in vivo. We show here that LTK63 intrapulmonary administration induced in the mouse lung a specific gene expression signature characterized by the up-regulation of cell cycle genes, several host defense genes, chemokines, chemokine receptors, and immune cell-associated genes. Such a transcriptional profile reflected the activation of alveolar macrophages and the recruitment to the lung of T and B cells and innate immune cells such as granulocytes, NK, and dendritic cells. All of these events were T cell dependent and specific for LTK63 because they were absent in SCID and nude mice. Additionally, we showed that LTK63 induces a potent adaptive immune response against itself directed to the lung. We propose that acquired response to LTK63 is the driving force for the local recruitment of both adaptive and innate immune cells. Our data suggest that LTK63 acts as an airway infection mimic that establishes a generic protective environment limiting respiratory infection by innate immune mechanisms and by improving adaptive responses to invading pathogens.
- Published
- 2007
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30. Reduced number and function of peripheral dendritic cells in coeliac disease.
- Author
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Ciccocioppo R, Ricci G, Rovati B, Pesce I, Mazzocchi S, Piancatelli D, Cagnoni A, Millimaggi D, Danova M, and Corazza GR
- Subjects
- Adolescent, Adult, Aged, Celiac Disease diet therapy, Cell Count, Cell Differentiation immunology, Cytokines metabolism, Female, Flow Cytometry methods, Gliadin administration & dosage, Humans, Immunophenotyping, Lymphocyte Culture Test, Mixed, Male, Middle Aged, Celiac Disease immunology, Dendritic Cells immunology
- Abstract
Dendritic cells (DC) play a pivotal role in shaping the immune response in both physiological and pathological conditions. In peripheral blood at least two subsets, the myeloid and plasmacytoid, have been described as having different T stimulatory functions and a variable degree of maturation. Certainly, antigen presentation plays a crucial role in the pathogenesis of coeliac disease and circulating immune cells are thought to reflect the state of immune response within the gut. Therefore, we aimed to investigate the quantitative and phenotypical modifications of peripheral blood DC, together with their functional properties, in this pathological condition. Blood samples from 11 untreated patients before and after a course of gluten-free diet, 27 treated patients and 14 controls underwent flow-cytometric analysis, while immunomagnetically sorted DC from the CD patients and eight human leucocyte antigen (HLA)-DQ2/8(+) bone marrow donors were used to evaluate maturation status through the CD83 expression, cytokine profile for interleukin (IL)-6, IL-10, IL-12 and interferon (IFN)-alpha by enzyme-linked immunosorbent assay (ELISA), and functional properties by mixed leucocyte reaction before and after pulsing with digested gliadin. We found that in both untreated and treated patients, a significant reduction of the entire DC population, mainly the plasmacytoid subset, in comparison to healthy controls was observed. In active disease, an impaired allogenic lymphocyte reaction and a significant reduction of IFN-alpha production, paralleled by the presence of a more immature status, were also demonstrated. All the latter modifications have been reverted by pulsing DC with digested gliadin.
- Published
- 2007
- Full Text
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31. Expression of cytokines, inducible nitric oxide synthase, and matrix metalloproteinases in pouchitis: effects of probiotic treatment.
- Author
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Ulisse S, Gionchetti P, D'Alò S, Russo FP, Pesce I, Ricci G, Rizzello F, Helwig U, Cifone MG, Campieri M, and De Simone C
- Subjects
- Acute Disease, Cytokines analysis, Humans, Matrix Metalloproteinases analysis, Nitric Oxide Synthase analysis, Pouchitis enzymology, Pouchitis immunology, Cytokines biosynthesis, Matrix Metalloproteinases metabolism, Nitric Oxide Synthase metabolism, Pouchitis therapy, Probiotics therapeutic use
- Abstract
Objective: The efficacy of probiotic organisms in the treatment of pouchitis has been reported. In the present study, we evaluated the tissue levels of pro- and anti-inflammatory cytokines, nitric oxide synthase, and matrix metalloproteinases in control and inflamed pouches before and after antibiotic and probiotic treatment of patients with acute pouchitis., Methods: Pouch biopsy samples were obtained from seven patients with pouchitis before and after antibiotic and probiotic treatment. Tissue samples from five patients with normal pouches were used as controls. Cytokines were determined by ELISA, matrix metalloproteinase activity was evaluated by zymograms, and nitric oxide synthase activity was determined by measuring arginine to citrulline conversion., Results: Tissue levels of tumor necrosis factor a increased (p < 0.01) in pouchitis relative to uninflamed pouches and reduced after antibiotic and probiotic treatment. Also, interferon y and interleukin 1alpha (IL-1alpha) augmented in pouchitis, but their increase did not reach statistical significance. The latter, however, were lower (p < 0.05) after treatment with the antibiotics and probiotics. Tissue levels of IL-4 and IL-10 were unchanged in inflamed pouches and unaffected by antibiotic treatment. However, IL-10 increased (p < 0.05) after probiotic treatment. Moreover, inflamed pouches had higher levels of inducible nitric oxide synthase and gelatinase activities, which decreased after treatment., Conclusions: The ability of antibiotic and probiotic treatments to increase tissue levels of IL-10, at a higher level than those observed in control pouches, and to decrease, to levels present in control pouches, proinflammatory cytokine, inducible nitric oxide synthase, and matrix metalloproteinase activity may suggest a mechanism of action to explain the efficacy of this therapeutic regime in pouchitis.
- Published
- 2001
- Full Text
- View/download PDF
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