PHENOTYPIC SWITCHING CHANGES ADHESION PROFILE OF Candida tropicalis MORPHOTYPES IN HELA CELLS

Candida tropicalis has been frequently associated with candidemia cases, being considered a important human pathogen, expressing many virulence factors, as adhesion and phenotypic switching event. Phenotypic switching have being related to increased of yeast adaptability, can changes the yeast's ability to colonize and adhere to the host. This research, aim avalueted the morphotype adhesion profile of the C. tropicalis switching systems 49.07, 100.10 and 335.07 in HeLa cells. After, HeLa cells (ATCC, CCL2TM) achieving 80% confluence in monolayer, it was co-cultured with C. tropicalis morphotypes (1x107cells/ml) and incubated for 2 hours at 37 ° C in 5% CO2, in DMEM medium. The wells were washed with PBS buffer, staining with crystal violet (1%) and washed with PBS. Ethanol:acetone solution was added to remove color from epithelial cells. The absorbance was measure at 570 nm, for quantification total biomass. To evaluate the mean of yeasts adhered, HeLA cells was adhered in glass coverslips in cell culture plates, and co-cultivation happened as described. Subsequently, coverslips was fixed and staining by methylene blue (1%), and observed in optical microscope. The quantification by total biomass, evidenced adhesion ability for all morphotypes in HeLA cells. Of the ten switching lines evaluated, 70% differed from parental adhesion. The parental morphotype of 335.07 system showed high adhesion, in opposition to Irregular center revertant (p<0.001). Both variants of 100.10 system (Crepe and Rough) differed from parental adhesion, as the Rough revertent (p <0.001). The Rough, Crepe and Rough revertents, of system 49.07, were more adherent than parental (p <0.001). The means of yeasts adhered in HeLa cells by morphotypes from 49.07, was similar between variants and Parental, however, Crepe revertent (5,944473 yeast/HeLa) and Rough revertent (6,531272 yeast/HeLa) differed from Parental (4,135222 yeast/HeLa). Crepe of 100.10, showed a small number of yeasts adhered in HeLa (4,158214 yeast/HeLa) than parental (5,404858 yeast/HeLa), while Revertants presenting similar means of adhered yeasts per HeLa cells, differ from their variants. Irregular center of 335.07, presented yeast mean per HeLa higher than was observed in its Revertent. The total biomass data and the mean of adhered yeasts, presented high correlation, by the Pearson correlation coefficient. Considering these results, it is possible observed a tendency of the variants for a low number of yeasts adhered per HeLa, indicating what colony complexity is not a factor related to adhesion, and what phenotypic switching in C. tropicalis is associated with changes the adhesion profile in HeLa cells.