Identification of genes of Saccharomyces cerevisiae envolved im immunomodulation using reporter intestinal epithelial cells

ROMANIN D E; PERELMUTER K.,; CARBO N., ; GARROTE G L; AGUILAR P.S.; BOLLATI-FOGOLIN M.; RUMBO M

Otro; LXII Reunión Anual de La Sociedad Argentina de Inmunología; 2014

Sociedad Argentina de Inmunología

In previous works we have demonstrated the ability of different yeast to modulate the inflammatory response triggered by diverse innate immunity agonists in vitro. This capacity is dependent on yeast viability. In the present work Saccharomyces cerevisiae single-gene mutants were used to evaluate the role of those genes in the immunomodulatory effect. We used a stably transfected cell line that reports epithelial inflammation based on human intestinal enterocyte HT29 cells with GFP as reporter driven by a NF-kB dependent artificial promoter. From a collection of approximately 5000 non-essential gene deletion mutants, forty were selected according to their described phenotypes in Saccharomyces Genome Database (SGD). The selection of mutants was focused 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 yeast YER044C, with a deletion on gene ERG28 coding for an enzyme located in the endoplasmic reticulum which is involved in ergosterol biosynthesis, shows a significant lower immunomodulatory capacity compared to the parental haploid strain By4741, using TNF-a as proinflammatory agonist (p