Capacity of Shiga toxin to induce in vitro neutrophil extracellular traps (NETs)

RAMOS, MV; MEJIAS, MP; SABBIONE, F; ABREY-RECALDE, MJ; PANEK, CA; TREVANI, A; PALERMO, MS

Ouro Preto, Minas Gerais

Simposio; Keystone Symposia on mollecular and cellular biology: The Innate Immune Response in the Pathogenesis of Infectious Disease.; 2013

Key Stone,Universidade Federal de Ouro Preto (UFOP)

Capacity of Shiga toxin to induce in vitro neutrophil extracellular traps (NETs). Ramos, MV; Mejias, P; Sabbione, F; Abrey-Recalde, MJ; Panek, A; Trevani, A; Palermo, MS. Hemolytic Uremic Syndrome (HUS), a vascular disease characterized by hemolityc anemia, thrombocitopenia and acute renal failure is caused by enterohemorraghic Shiga toxins(Stx)-producing bacteria. Besides Stx, inflammatory response mediated by neutrophils (PMN) is essential to HUS evolution. The aim of this work was to determine the Stx direct effect on PMN and the possible induction of neutrophil extracellular traps (NETs). For this purpose PMN were purified from peripheral blood of healthy donors. Then they were incubated with Stx and the surface expression of activation markers such as CD11b and CD66b was evaluated by flow cytometry. Both markers were increased following stimulation. (MFI±SE: CD66b: basal=65±11, Stx=154±11; CD11b: basal=471±48, Stx=619±100). We also evaluated the capacity of Stx to induce on PMN the extrusion of NETs. Isolated PMN were incubated with phorbol myristate acetate (PMA), a positive inductor of netosis, or Stx and cells stained with PI and anti-elastase antibody were counted by confocal microscopy (Basal=0, PMA=5.6*± 1, Stx=2.8±0.24*; n=3, *p