Gene transcription studies ofCandida albicansfollowing infection of HEp2 epithelial cells

Abstract

Previously we observed that infection of HEp2 epithelial cells with Candida albicans results in HEp2 cell actin rearrangement as well as reduced membrane ruffling and motility and that supernatants of a C. albicans culture (Candida metabolite) caused the same changes. In this study, we used microarray analysis to determine changes in gene transcription of C. albicans following infection of HEp2 cells compared to control cultures grown in the absence of HEp2 cells. We observed 201 genes whose regulation was increased at least 2-fold following a 3 h incubation with HEp2 cells as well as 87 genes that are down-regulated. Among the up-regulated genes were ALS2 and ALS5 both of which encode proteins that provide an adherence function for C. albicans. To confirm the changes in ALS transcription, we measured by RT-PCR ALS1-9 at 1 h intervals for a total of 4 h. After 1 h of infection, several of the ALS genes were up-regulated compared to C. albicans grown alone. At 2-4 h, an increase in most of the ALS genes was observed in both infected and control cultures. ALS7 transcription was observed only at 3-4 h, but transcription was similar in both infected and control cultures. By RT-PCR, ALS2 and 5, similar to the microarray data, were significantly increased in infected cells at 3 h. Our results show that gene transcription following the adherence of C. albicans to HEp2 cells includes the up-regulation of genes encoding members of a family of known host recognition adhesins that may be critical to successful colonization and invasion of the organism.