Limited expression of TLR9 on T cells and its functional consequences in patients with nonalcoholic fatty liver disease

We demonstrated a limited expression of Toll-like receptor 9 (TLR9) and interferon gamma (IFN-γ) production by T cells in patients with simple steatosis (SS). The limited expression of TLR9 on T cells was directly associated with lower liver necroinflammatory activity and fibrosis, and lower antrophometric and biochemical alterations of nonalcoholic fatty liver disease. Besides, co-stimulation of T cell activation via TLR9 induced a limited number of IFN-γ-producing CD8+ T cells in SS patients. Accordingly, these patients showed a low frequency of circulating type 1 CD8+ cells whereas additional pro-inflammatory signals may be responsible for the higher frequency observed at baseline in the context of steatohepatitis.Fil: 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: 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: Billordo, Luis Ariel. 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: Ameigeiras, Beatriz. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Poncino, Daniel. Sanatorio Municipal Dr. Julio Méndez. Servicio de Gastroenterología; ArgentinaFil: Benavides, Javier. Hospital Británico de Buenos Aires; ArgentinaFil: Colombato, Luis. Hospital Británico de Buenos Aires; 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

“Hepatic toxicity by methotrexate with weekly single doses associated with folic acid in rheumatoid and psoriatic arthritis. What is its real frequency?”

Introduction and Objectives: Liver injury caused by methotrexate (MTX) has mostly been investigated without applying criteria for the assessment of causality of drug induced liver injury (DILI). Hence, the existence of DILI by MTX in many cases is debatable. This study aimed to describe the frequency and characteristics of liver injury caused by MTX, applying DILI diagnostic criteria. Material and methods: Rheumatoid arthritis (RA) and psoriatic arthritis (PsA) patients who were treated with MTX in association with folic acid were included. Serial determinations of alanine amino transferase (ALT) and aspartate amino transferase (AST) were performed. The Roussel Uclaf Causality Assessment Method (RUCAM) was applied in cases of increases of ALT/AST over 1.5 upper limit of normal. Liver biopsy was considered when the total cumulative dosage (TCD) of MTX was ≥3.5 g. Results: A total of 43 patients were analyzed (median follow up 32 (range: 1–48) months; 3.33 ALT/AST determinations per year). Five subjects presented an increase of ALT/AST. All presented a RUCAM score for MTX ≤ 2 (improbable). Three had a RUCAM score for non-steroidal anti-inflammatory drugs ≥7 (probable) and two patients presented non-alcoholic fatty liver disease. Five patients with no other cause for liver disease consented to liver biopsy (TCD MTX: median 5.1; range: 3.5–7.4 g). No significant fibrosis or steatosis was evident on histology. Conclusions: No biochemical or significant histological liver toxicity for MTX was demonstrated when applying causality criteria for DILI. More studies with this methodology are necessary in order to improve the assessment of its frequency

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