In Vivo Imaging of Trypanosome-Brain Interactions and Development of a Rapid Screening Test for Drugs against CNS Stage Trypanosomiasis
| Autor: | Martin C. Taylor, Michael P. Barrett, Jonathan A. Coles, James M. Brewer, Jeremy C. Mottram, Ryan Ritchie, Alex P. McLatchie, Jean Rodgers, Elmarie Myburgh, Peter G. E. Kennedy |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Drug Evaluation
Preclinical Pathogenesis Parasitemia Melarsoprol Protozoology Mice Diminazene 0302 clinical medicine Infectious Diseases of the Nervous System Drug Discovery Image Processing Computer-Assisted African trypanosomiasis Luciferases 0303 health sciences biology lcsh:Public aspects of medicine Brain 3. Good health Host-Pathogen Interaction Infectious Diseases Host-Pathogen Interactions Medicine Female Research Article Neglected Tropical Diseases medicine.drug Trypanosoma Drugs and Devices lcsh:Arctic medicine. Tropical medicine Drug Research and Development Infectious Disease Control lcsh:RC955-962 Trypanosoma brucei brucei 030231 tropical medicine Antiprotozoal Agents Trypanosoma brucei Microbiology African Trypanosomiasis 03 medical and health sciences Trypanosomiasis Parasitic Diseases medicine Animals Bioluminescence imaging Biology 030304 developmental biology Staining and Labeling Public Health Environmental and Occupational Health lcsh:RA1-1270 biology.organism_classification medicine.disease Virology Disease Models Animal Microscopy Fluorescence Multiphoton Immunology Parastic Protozoans Parasitology Infectious Disease Modeling |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Vol 7, Iss 8, p e2384 (2013) |
| ISSN: | 1935-2727 1935-2735 |
| Popis: | Human African trypanosomiasis (HAT) manifests in two stages of disease: firstly, haemolymphatic, and secondly, an encephalitic phase involving the central nervous system (CNS). New drugs to treat the second-stage disease are urgently needed, yet testing of novel drug candidates is a slow process because the established animal model relies on detecting parasitemia in the blood as late as 180 days after treatment. To expedite compound screening, we have modified the GVR35 strain of Trypanosoma brucei brucei to express luciferase, and have monitored parasite distribution in infected mice following treatment with trypanocidal compounds using serial, non-invasive, bioluminescence imaging. Parasites were detected in the brains of infected mice following treatment with diminazene, a drug which cures stage 1 but not stage 2 disease. Intravital multi-photon microscopy revealed that trypanosomes enter the brain meninges as early as day 5 post-infection but can be killed by diminazene, whereas those that cross the blood-brain barrier and enter the parenchyma by day 21 survived treatment and later caused bloodstream recrudescence. In contrast, all bioluminescent parasites were permanently eliminated by treatment with melarsoprol and DB829, compounds known to cure stage 2 disease. We show that this use of imaging reduces by two thirds the time taken to assess drug efficacy and provides a dual-modal imaging platform for monitoring trypanosome infection in different areas of the brain. Author Summary Trypanosoma brucei, a parasite transmitted by the bite of tsetse fly, is responsible for the disease human African trypanosomiasis (HAT). In advanced stages of HAT, trypanosomes invade the central nervous system (CNS), resulting in an array of neurological symptoms, and eventually death. Existing drugs for treatment of HAT are highly unsatisfactory and new safe drugs are urgently needed. Currently, potential drugs for HAT are screened in a mouse model that relies on the emergence of trypanosomes from tissues and their detection in blood. This can take up to 200 days, making selection and further development of new drugs slow and costly. Here, we employ in vivo imaging and genetically modified trypanosomes to monitor parasite distribution throughout the body in live infected mice. Our bioluminescence imaging approach provides sensitive detection of trypanosomes at sites of infection, allowing more rapid and more effective in vivo screening of candidate HAT drugs. Higher resolution intra-vital microscopy was used to investigate trypanosome dynamics in the brain and their accessibility to drugs during infection. These approaches allow more sensitive real time tracking of trypanosomes during chronic infections and will provide new insights about trypanosome pathogenesis in future experiments. |
| Databáze: | OpenAIRE |
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