Bioluminescent Imaging of Trypanosoma brucei Shows Preferential Testis Dissemination Which May Hamper Drug Efficacy in Sleeping Sickness

Abstract

Monitoring Trypanosoma spread using real-time imaging in vivo provides a fast method to evaluate parasite distribution especially in immunoprivileged locations. Here, we generated monomorphic and pleomorphic recombinant Trypanosoma brucei expressing the Renilla luciferase. In vitro luciferase activity measurements confirmed the uptake of the coelenterazine substrate by live parasites and light emission. We further validated the use of Renilla luciferase-tagged trypanosomes for real-time bioluminescent in vivo analysis. Interestingly, a preferential testis tropism was observed with both the monomorphic and pleomorphic recombinants. This is of importance when considering trypanocidal drug development, since parasites might be protected from many drugs by the blood-testis barrier. This hypothesis was supported by our final study of the efficacy of treatment with trypanocidal drugs in T. brucei-infected mice. We showed that parasites located in the testis, as compared to those located in the abdominal cavity, were not readily cleared by the drugs. Human African trypanosomiasis or sleeping sickness, caused by two subspecies of Trypanosoma brucei, is endemic in Subsaharan Africa. There is no vaccine and the currently used drugs are toxic and can cause severe side effects and even death. At present, we do not know how and when parasites can leave the blood and penetrate into organs (especially the brain). Such knowledge will be very helpful to develop and validate new drugs that can clear the parasite from both the blood and the tissues. In this study, we developed a novel technique allowing us to track the parasites in a live animal by the use of light signals. By following the luminescent parasites in the mouse we showed that, interestingly, the organisms migrate very early in infection to the testes. Here, they may be protected from the immune system and from drugs. Indeed when treating the mice with a sub-optimal dose of medicine, the parasites in this location were not cleared and subsequently could cause a reinvasion into the blood of the host.