IDENTIFICATION OF GENES OF SACCHAROMYCES CEREVISIAE INVOLVED IN IMMUNOMODULATION USING REPORTER INTESTINAL EPITHELIAL CELLS

ROMANIN, D; PERELMUTER, K; CARBÓ, N; GARROTE, G; AGUILAR, P.; BOLLATI, M; RUMBO, M

Mar del Plata

Congreso; LXII Reunión científica anual SAI; 2014

Sociedad Argentina de Inmunología

In previous works we have demonstrated the ability of different yeast to modulate the inflammatory response triggered by diverseinnate immunity agonists in vitro. This capacity is dependent on yeast viability. In the present work Saccharomyces cerevisiaesingle-gene mutants were used to evaluate the role of those genes in the immunomodulatory effect. We used a stably transfectedcell line that reports epithelial inflammation based on human intestinal enterocyte HT29 cells with GFP as reporter driven by aNF-kB dependent artificial promoter. From a collection of approximately 5000 non-essential gene deletion mutants, forty wereselected according to their described phenotypes in Saccharomyces Genome Database (SGD). The selection of mutants wasfocused on genes involved in yeast metabolism, cell wall synthesis, protein glycosilation and membrane transport.Most of the mutants analyzed showed no altered capacity to modulate epithelial response activation. However, the mutant yeastYER044C, with a deletion on gene ERG28 coding for an enzyme located in the endoplasmic reticulum which is involved inergosterol biosynthesis, shows a significant lower immunomodulatory capacity compared to the parental haploid strain By4741,using TNF-a as proinflammatory agonist (p<0,05, ANOVA + Bonferroni?s multiple comparison test). On the other hand, two mutantswith significantly higher modulation capacity were identified: YGR032W and YJL078C, with mutations on the catalytic subunit forthe beta 1,3 glucan synthase (coded in the gene GSC2) and a protein coded by the gene PRY3 which participates in the transport ofacylsterols respectively.Together these results set the bases for the study of the molecules present in yeast that are responsible for the interactionbetween microorganism and epithelial cells and the mechanistic basis of epithelial proinflammatory modulation.