IMEX   05356
INSTITUTO DE MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Increased levels of Neutrophil extracellular traps in HUS patients
Autor/es:
RAMOS, MARIA VICTORIA; MEJIAS, MARIA PILAR; SABBIONE FLORENCIA; FERNANDEZ BRANDO, R; SANTIAGO, ADRIANA; AMARAL, MARIA MARTA; EXENI, RAMON; TREVANI, ANALIA; PALERMO, MARINA SANDRA
Lugar:
Boston
Reunión:
Congreso; 9th International Symposium on Shiga Toxin (Verocytotin)-Producing Escherichia coli Infections . VTEC2015; 2015
Resumen:
Title: Increased levels of Neutrophil extracellular traps in HUS patients Authors: María Victoria Ramos1, María Pilar Mejias1, Florencia Sabbione2, Romina Jimena Fernandez-Brando1, Adriana Patricia Santiago3, María Marta Amaral4, Ramón Exeni3, Analia Trevani2 and Marina Sandra Palermo1. 1Laboratorio de Patogénesis e Inmunología de Procesos Infecciosos and 2Laboratorio de Inmunidad Innata, Instituto de Medicina Experimental Medicine (IMEX-CONICET), Academia Nacional de Medicina, Buenos Aires, Argentina. 3Departamento de Nefrología, Hospital Municipal del Niño, San Justo, Provincia de Buenos Aires, Argentina. 4Laboratorio de Fisiopatogenia, Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina. Introduction: Hemolytic Uremic Syndrome (HUS), is a vascular disease caused by Shiga toxin (Stx) but also inflammatory response mediated by neutrophils (PMN) is essential for its evolution. PMN can release "neutrophil extracellular traps" (NETs) composed of DNA, histones and proteins such as elastase and myeloperoxidase. Since NETs are involved in several inflammatory diseases, the aim of this work was to investigate the presence of NETs in HUS patients. Methods: Peripheral blood from acute HUS and healthy children (HC)(n=15) was collected, plasmas were separated and PMN were isolated by gradients. PMN were incubated with medium or phorbolmyristate acetate (PMA) or monosodium-urate crystals (MSU), as inducers of NETosis. After 4h, free-DNA concentration and elastase activity were measured in the supernatants by fluorometry and spectrophotometry respectively. Circulating-free DNA (cf-DNA) in plasma was measured by fluorometry. Colocalization of cf-DNA and myeloperoxidase in plasma was analyzed by immunofluorescence and confocal microscopy. To evaluate the effect of NETs on endothelium, Human Glomerular Endothelial Cells (HGEC) were incubated during 24h with NETs, then cell viability was evaluated by spectrophotometry and cytokine levels in the supernatants were evaluated by ELISA. Results and Discussion: Plasmatic cf-DNA levels were increased in plasma from HUS compared to HC (HC=5,1±0,5; HUS=10±0,5*, Mean±SEM, ug/ml, n=15, *p