Alterations in response to resistin induced oxidative stress in circulating leukocytes from patients with nonalcoholic fatty liver disease (NAFLD)

We previously demonstrated that Resistin (RES) decrease reactive oxygen species levels (ROS) in peripheral blood mononuclear cells (PBMC) in non-NAFLD, but not in NAFLD patients. To further evaluate RES effects on oxidative stress and on antioxidant defenses (Aox) in PBMC from NAFLD patients, PBMC were cultured in presence/absence of RES (20 ng/ml) for 24h. PBMC from NAFLD patients had higher baseline ROS levels (1353 ± 498 vs. 468 ± 89, vs. non-NAFLD cells). In addition, PBMC from NAFLD patients showed lower superoxide dismutase (SOD) and glutathione reductase (GRd) activities, and higher glutathione peroxidase activity (p <0.05) than non-NAFLD PBMC. 24h incubation with RES induced an increase in catalase and GRd activities (p<0.01), and a decrease in SOD activity (p< 0.05) and glutathione content (p<0.05) in non-NAFLD PBMC. However, RES could not modulate enzymatic activities in PBMC from NALFD patients. In conclusion, RES presence favored the decrease in ROS in non-NAFLD PBMC by modulating Aox enzymes and GSH redox state, but not in PBMC from NAFLD patients. Redox signaling alterations would favor the inflammatory state and the progression of NALFD. The regulatory mechanisms controlled by Res could be used as potential therapeutic targets in NAFLD patients.Fil: García, Cecilia Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Piotrowsky, Barbara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Baz, Placida. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Benavidez, Javier. Hospital Británico de Buenos Aires; ArgentinaFil: Colombato, Luis. Hospital Británico de Buenos Aires; ArgentinaFil: Poncino, Daniel Alberto. Sanatorio Dr. “Julio Méndez”; ArgentinaFil: García, Daniel. Sanatorio Dr. “Julio Méndez”; ArgentinaFil: Fraga, César Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Cherñavsky, Alejandra Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina2019 Annual Meeting “Redox homeostasis: from signaling to damage”FerraraItaliaSociety for Free Radical Research Europ

A preliminar analysis of the influence of Resistin on immunological cells in nonalcoholic fatty liver disease

Background: Resistin (RES) is a cytokine highly expressed in plasma of nonalcoholic fatty liver disease (NAFLD) patients which promotes insulin resistance. It has a pro-inflammatory role through simulation of liver/adipose infiltrating monocytes/macrophages. As these cells are the main source of RES, a pro-inflammatory loop is created due to its role as chemokine for T cells. RES binding to adenylyl cyclase-associated protein 1 (CAP1), which expression was only reported in monocytes, activates nuclear factor kappa B (NF-kB). NF-kB is also activated via TCR to modulate CD69 expression in immunological cells. Aims: to evaluate CAP1 expression in peripheral blood mononuclear cells (PBMC) from patients and controls (Co), and RES-mediated modulation of CD69. Methods: PBMC were obtained by density gradient from whole blood of NAFLD patients (n= 8) and Co (n= 10). All patients provided written informed consent. CAP1 expression was studied by flow cytometry (FC) in PBMC stained with anti-CD3, -CD4, -CD14 and -CD56 antibodies. For functional approach, PBMC from Co were stimulated with coated anti-CD3 (3ug/ml) +/- RES (500 ng/ml) for 24 h, stained with anti-CD3, -CD4 and -CD56 and evaluated for CD69 expression by FC. Mann-Whitney test was used. Results: CAP1 expression is decreased in monocytes (p=0.018), CD3+CD4+ (p=0.016) and CD3+ CD4- (p=0.018) cells in patients with NAFLD (vs. Co). RES alone did not activate PBMC. Costimulation with anti-CD3+RES decreased CD69 mean fluorescence intensity (MFI) in CD3+CD4+ and CD3+CD4-, natural killer T (NKT) and NK CD56 bright cells (p=0.0043, p=0.021, p=0.044 and p=0.035 respectively; anti-CD3+RES vs. anti-CD3). Conclusion: CAP1 is a functional receptor in healthy donors able to regulate RES influence on immunological cells. Even though RES does not induce CD69 expression per se, it can modulate the activation of immunological cells. Thus, an overall decrease in CAP1 expression might modulate the stimulus induced by RES in the context of NAFLD. This potential regulatory circuit deserves further investigation.Fil: Garcia, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Alegre, Nadia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Baz, Placida. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Benavidez, Javier. Hospital Británico de Buenos Aires; ArgentinaFil: Colombato, Luis. Hospital Británico de Buenos Aires; ArgentinaFil: Poncino, Daniel Alberto. Sanatorio Méndez; ArgentinaFil: Garcia Daniel. Hospital Británico de Buenos Aires; ArgentinaFil: Romeo, Juan Manuel. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Ameigeiras, Beatriz. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Cherñavsky, Alejandra Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaLXI Reunión Anual de la Sociedad Agentina de Investigación Clínica; LXIV Reunión Anual de la Sociedad Argentina de Inmunología; XLVIII Reunión Anual de la Sociedad Argentina de Farmacología Experimental; VII Reunión Anual de la Sociedad Argentina de Nanomedicina y V Congreso Nacional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de InmunologíaSociedad Argentina de Investigacion ClínicaSociedad Argentina de Farmacología ExperimentalSociedad Argentina de NanomedicinaAsociación Argentina de Ciencia y Tecnología de Animales de Laboratori

A Decreased Response to Resistin in Mononuclear Leukocytes Contributes to Oxidative Stress in Nonalcoholic Fatty Liver Disease

Background: Deregulation of immune response and oxidative stress contribute to nonalcoholic fatty liver disease (NAFLD) pathogenesis. Resistin is a physiological modulator of inflammation and redox homeostasis of different cell types. Increased resistin serum concentration and the direct association between resistin hepatic expression and NAFLD severity suggest that resistin participates in NAFLD pathogenesis. Aims: To evaluate resistin-induced regulation of redox homeostasis in mononuclear leukocytes from NAFLD patients and controls. Methods: We evaluated basal and resistin-mediated modulation of reactive oxygen species (ROS) and glutathione content by flow cytometry, and antioxidant enzyme activities by spectrophotometry. Results: Peripheral blood mononuclear cells (PBMC) from NAFLD patients showed higher ROS content and glutathione peroxidase activity and lower glutathione content, superoxide dismutase and glutathione reductase activities than control PBMC. Resistin decreased ROS levels and superoxide dismutase activity and increased glutathione reductase and catalase activities in PBMC from controls but not from patients. Resistin decreased glutathione content in PBMC from control and NAFLD patients, with greater effect on patient cells. Basal and resistin-modulated ROS levels were directly associated with obesity-related risk factors for NAFLD. Hepatic myeloid cells and T-lymphocytes from NAFLD patients showed higher basal ROS content than cells from controls. Resistin decreased ROS levels in hepatic T-lymphocytes from controls but not from patients. Conclusions: Resistin regulates redox homeostasis in mononuclear leukocytes. A decreased response to resistin in leukocytes from NAFLD patients is associated with an impaired redox homeostasis.Fil: García, Cecilia Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Piotrkowski, Barbara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Baz, Placida. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: Poncino, Daniel Alberto. Sanatorio Dr. Julio Méndez; ArgentinaFil: Benavides, Javier. Hospital Británico de Buenos Aires; ArgentinaFil: Colombato, Luis. Hospital Británico de Buenos Aires; ArgentinaFil: Reyes Toso, María Laura. Fundación Favaloro; ArgentinaFil: Yantorno, Silvina. Fundación Favaloro; ArgentinaFil: Descalzi, Valeria. Fundación Favaloro; ArgentinaFil: Gondolesi, Gabriel Eduardo. Fundación Favaloro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fraga, César Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Cherñavsky, Alejandra Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentin

Ombitasvir/paritaprevir/ritonavir/dasabuvir ± ribavirin is safe and effective in HCV-infected patients in a real-life cohort from Latin America

Information about the use of ombitasvir/paritaprevir/ritonavir/dasabuvir ± ribavirin (OBV/PTV/r/DSV ± RBV) in real-clinical practice in Latin America is scarce. We aimed to confirm safety and effectiveness of OBV/PTV/r/DSV ± RBV therapy in real-world setting. We analyzed a cohort of patients with genotype 1 infection treated with OBV/PTV/r/DSV ± RBV. Data on demographics, clinical features, safety, and virological response were retrospectively collected from 21 centers in Latin America. A total of 96 patients received OBV/PTV/r/DSV, associated with RBV in 68% of the cases. Most were genotype 1b (80%), 56 (58%) had cirrhosis, and 45 (47%) failed prior HCV treatment. Adverse events occurred in 62% of patients. The most common adverse events were pruritus (21%), hyperbilirubinemia (17%), and asthenia (17%). Five patients discontinued therapy prematurely due to hepatic decompensation, three of them were Child-Pugh B at baseline and one patient died due to multi-organ failure. Follow up HCV-RNA 12 weeks after completion of therapy was evaluated in all the patients and sustained virologic response rate was 97%. No virologic breakthrough was detected. Our study confirms that OBV/PTV/r/DSV treatment is highly effective in patients with chronic HCV without cirrhosis or with Child-Pugh A cirrhosis in non-European populations. Adverse events were often mild and rarely led to treatment discontinuation except for patients with Child-Pugh B cirrhosis or with previous history of hepatic decompensation. These results can support the development of public strategies to expand the access of OBV/PTV/r + DSV and other DAAs combinations in order to reduce the burden of HCV infection in our region.Fil: Mendizabal, Manuel. Hospital Universitario Austral; ArgentinaFil: Haddad, Leila. Hospital Italiano; ArgentinaFil: Gallardo, Patricia E.. Fundación Sayani; ArgentinaFil: Ferrada, Alejandro. Hospital Clinico San Borja Arriaran; ChileFil: Soza, Alejandro A.. Universidad Católica de Chile; Chile. Pontificia Universidad Católica de Chile; ChileFil: Adrover, Raul. Centro de Hepatología; ArgentinaFil: Aravena, Edmundo. Hospital Clinico San Borja Arriaran; ChileFil: Roblero, Juan P.. Hospital Clinico San Borja Arriaran; ChileFil: Prieto, Jhon. Clínica Universitaria Colombia y Centro de Enfermedades Hepáticas y Digestivas; ColombiaFil: Vujacich, Claudia. Fundacion Centro de Estudios Infectologicos; ArgentinaFil: Romero, Gustavo. Gobierno de la Ciudad de Buenos Aires. Hospital de Gastroenterología ; ArgentinaFil: Muñoz, Alberto. Gobierno de la Ciudad de Buenos Aires. Hospital de Gastroenterología ; ArgentinaFil: Anders, Margarita. Hospital Alemán; ArgentinaFil: Hernández, Nelia. Hospital de Clinicas Dr. Manuel Quintela; UruguayFil: Coccozella, Daniel. Centro de Hepatología; ArgentinaFil: Gruz, Fernando. Fundación Favaloro; ArgentinaFil: Reggiardo, Maria V.. Provincia de Santa Fe. Ministerio de Salud y Medio Ambiente - Rosario. Hospital Provincial del Centenario; ArgentinaFil: Ruf, Andres E.. FUNDIEH; ArgentinaFil: Varón, Adriana. Instituto de Cardiologia; ColombiaFil: Cartier, Mariano. Gobierno de la Ciudad de Buenos Aires. Hospital de Gastroenterología ; ArgentinaFil: Pérez Ravier, Roberto. Instituto Universidad Escuela de Medicina del Hospital Italiano; ArgentinaFil: Ridruejo, Ezequiel. Hospital Universitario Austral; Argentina. Centro de Educación Medica E Invest.clinicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Peralta, Mirta. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas ; ArgentinaFil: Poncino, Daniel. Sanatorio Municipal Dr. Julio Méndez; ArgentinaFil: Vorobioff, Julio. Universidad Nacional de Rosario; ArgentinaFil: Aballay Soteras, Gabriel. Sanatorio Mitre; ArgentinaFil: Silva, Marcelo O.. Hospital Universitario Austral; Argentin