dsiRNA Inhibition of Tetraspanin-1 gene in Trichomonas vaginalis: Effect on host immune responses

Abstract

Trichomonas vaginalis (TV) is a common sexually transmitted infection, that causes Trichomoniasis. Little research is done to understand the mechanisms involved in TV’s communication with host cells. TV can communicate with host cells via specialized extracellular vesicles known as exosomes. Exosomes are membrane-bound, endocytic vesicles (MVBs) that are used to traffic proteins and genetic material, to neighboring cells. The sorting of proteins in the MVBs is facilitated by a group of proteins known as tetraspanins. The tetraspanins take part in a wide range of functions. TV genome contains several tertaspanin genes. Tetraspanin-1 (TSPAN-)1 is known to be lining the MVBs when TV attaches to host cells. To understand the role of TSPAN-1 gene in eliciting host immune responses, we designed custom dsiRNA primers to inhibit the expression of TSPAN-1 in TV parasites and study its effect on TV exosome formation and changes in TLR gene expression in host epithelial cells. We successfully inhibited TSPAN-1 gene in Trichomonas. The transfected parasites when exposed to host epithelial cells, resulted in reduced number of exosomes formed. In addition, TLR 2 gene expression was downregulated, while TLR 9 was upregulated when TSPAN-1 inhibited TVs were exposed to epithelial cells. TLR 4 gene expression, however, was not altered suggesting that tetraspanin-1 in the Trichomonas play a role in host -pathogen interactions and altering the host-immune responses in TV pathogenesis.