Suppression of mRNAs Encoding Tegument Tetraspanins from Schistosoma mansoni Results in Impaired Tegument Turnover

Abstract

Schistosomes express a family of integral membrane proteins, called tetraspanins (TSPs), in the outer surface membranes of the tegument. Two of these tetraspanins, Sm-TSP-1 and Sm-TSP-2, confer protection as vaccines in mice, and individuals who are naturally resistant to S. mansoni infection mount a strong IgG response to Sm-TSP-2. To determine their functions in the tegument of S. mansoni we used RNA interference to silence expression of Sm- tsp-1 and Sm- tsp-2 mRNAs. Soaking of parasites in Sm-tsp dsRNAs resulted in 61% (p = 0.009) and 74% (p = 0.009) reductions in Sm- tsp-1 and Sm- tsp-2 transcription levels, respectively, in adult worms, and 67%–75% (p = 0.011) and 69%–89% (p = 0.004) reductions in Sm- tsp-1 and Sm- tsp-2 transcription levels, respectively, in schistosomula compared to worms treated with irrelevant control ( luciferase) dsRNA. Ultrastructural morphology of adult worms treated in vitro with Sm- tsp-2 dsRNA displayed a distinctly vacuolated and thinner tegument compared with controls. Schistosomula exposed in vitro to Sm- tsp-2 dsRNA had a significantly thinner and more vacuolated tegument, and morphology consistent with a failure of tegumentary invaginations to close. Injection of mice with schistosomula that had been electroporated with Sm- tsp-1 and Sm- tsp-2 dsRNAs resulted in 61% (p = 0.005) and 83% (p = 0.002) reductions in the numbers of parasites recovered from the mesenteries four weeks later when compared to dsRNA-treated controls. These results imply that tetraspanins play important structural roles impacting tegument development, maturation or stability.

Author Summary

Schistosomes, or blood flukes, reside in the blood vessels surrounding the liver and bowel of their human hosts. They infect 200 million people and kill many thousands each year in developing countries. The parasites cover themselves in a unique series of cell membranes called the tegument. Molecules in the tegument membranes are a major target for the development of new drugs and vaccines against the parasite. Here we show that at least one member of a family of tegument membrane proteins called tetraspanins, Sm-TSP-2, is integral to the proper formation of the tegument and subsequent survival of the parasite in its human host, providing a potential mechanism by which a vaccine based on Sm-TSP-2 protects immunized hosts.