INVESTIGADORES
RUMBO Martin
congresos y reuniones científicas
Título:
Production of Reactive Oxigen Species by intestinal epithelial cells triggered by interaction with probiotic microorganisms
Autor/es:
ROMANIN, DAVID (1);; D GONZALEZ MACIEL,; HIRIART YANINA,; GARROTE, GRACIELA (1,3); MARTIN RUMBO
Reunión:
Congreso; 1st. French-Argentinean Congress of Immunology, November 2-5, 2010. Buenos Aires, Argentina.; 2010
Resumen:
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Different probiotic microorganisms may modulate
intestinal immune response, however the mechanisms involved remain poorly
characterized. Modulation of intestinal epithelial response may contribute to
probiotic function. Different intracellular signaling pathways have been
shown to be modulated by epithelial-microorganism interaction. Differential
targeting to the proteasome of key players of inflammatory pathways such as
IkBa protein can be modulated by probiotic bacteria by a mechanism dependent
on the production of Reactive Oxygen Species (ROS). The aim of the present
work was to set up a system to study this phenomena on intestinal epithelial
cells and test the effect on this pathway by a panel of probiotic bacteria
and yeast.
We standardized a procedure using Caco-2 human
intestinal epithelial cell line, a ROS-indicator dye (C-H2DCFDA) and
physiologic inducers of ROS production on Caco-2 cells to quantify the ROS
production by fluorescence microscopy. We used this system to test a panel of
20 strains from genus Lactococcus, Enterococcus and Lactobacillus and yeasts
from genus Saccharomyces and Kluyveromyces on the induction of ROS. The
capacity of these microorganisms to down-regulate the expression of CCL20 was
evaluated using a reporter CCL20-luc Caco-2 cell line and flagellin
stimulation. Some of the bacterial strains as Lactobacillus brevis ATCC 8287
and Enterococcus faecalis induced a significant two-fold production of
endogenous ROS. Although yeast strains did not induce ROS response, they
reduced more than 90% the flagellin-induced CCL20 response.
A system to evaluate the capacity of microorganisms to
trigger ROS production in intestinal epithelial cells was developed. No
correlation was observed between the capacity to modulate CCL20 response and
ROS production, indicating that other mechanisms triggered by
microorganism-epithelial interaction may control this innate activation of
epithelial cells.