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A panel of Trypanosoma brucei strains tagged with blue and red-shifted luciferases for bioluminescent imaging in murine infection models

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dc.contributor.author van Reet, N.
dc.contributor.author Van De Vyver, H.
dc.contributor.author Pyana, P. P.
dc.contributor.author Van der Linden, A. M.
dc.contributor.author Büscher, P.
dc.date.accessioned 2015-02-25T13:56:08Z
dc.date.available 2015-02-25T13:56:08Z
dc.date.issued 2014
dc.identifier.issn 1935-2727
dc.identifier.doi http://dx.doi.org/10.1371/journal.pntd.0003054
dc.identifier.other ITG-B1B
dc.identifier.other ITG-B3B
dc.identifier.other ITG-BLA
dc.identifier.other DBM
dc.identifier.other U-PARDIA
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other FTA
dc.identifier.other OAJ
dc.identifier.other E-only
dc.identifier.other Abstract
dc.identifier.other UPD57
dc.identifier.uri http://hdl.handle.net/10390/8299
dc.description.abstract BACKGROUND: Genetic engineering with luciferase reporter genes allows monitoring Trypanosoma brucei (T.b.) infections in mice by in vivo bioluminescence imaging (BLI). Until recently, luminescent T.b. models were based on Renilla luciferase (RLuc) activity. Our study aimed at evaluating red-shifted luciferases for in vivo BLI in a set of diverse T.b. strains of all three subspecies, including some recently isolated from human patients. METHODOLOGY/PRINCIPAL FINDINGS: We transfected T.b. brucei, T.b. rhodesiense and T.b. gambiense strains with either RLuc, click beetle red (CBR) or Photinus pyralis RE9 (PpyRE9) luciferase and characterised their in vitro luciferase activity, growth profile and drug sensitivity, and their potential for in vivo BLI. Compared to RLuc, the red-shifted luciferases, CBR and PpyRE9, allow tracking of T.b. brucei AnTaR 1 trypanosomes with higher details on tissue distribution, and PpyRE9 allows detection of the parasites with a sensitivity of at least one order of magnitude higher than CBR luciferase. With CBR-tagged T.b. gambiense LiTaR1, T.b. rhodesiense RUMPHI and T.b. gambiense 348 BT in an acute, subacute and chronic infection model respectively, we observed differences in parasite tropism for murine tissues during in vivo BLI. Ex vivo BLI on the brain confirmed central nervous system infection by all luminescent strains of T.b. brucei AnTaR 1, T.b. rhodesiense RUMPHI and T.b. gambiense 348 BT. CONCLUSIONS/SIGNIFICANCE: We established a genetically and phenotypically diverse collection of bioluminescent T.b. brucei, T.b. gambiense and T.b. rhodesiense strains, including drug resistant strains. For in vivo BLI monitoring of murine infections, we recommend trypanosome strains transfected with red-shifted luciferase reporter genes, such as CBR and PpyRE9. Red-shifted luciferases can be detected with a higher sensitivity in vivo and at the same time they improve the spatial resolution of the parasites in the entire body due to the better kinetics of their substrate D-luciferin. en_US
dc.language English en_US
dc.subject Protozoal diseases en_US
dc.subject Sleeping sickness en_US
dc.subject Trypanosomiasis, African en_US
dc.subject Trypanosoma brucei brucei en_US
dc.subject Trypanosoma brucei gambiense en_US
dc.subject Trypanosoma brucei rhodesiense en_US
dc.subject Vectors en_US
dc.subject Tsetse flies en_US
dc.subject Glossina morsitans morsitans en_US
dc.subject Bioluminescence en_US
dc.subject Luciferase en_US
dc.subject In vivo en_US
dc.subject Mice en_US
dc.subject Growth en_US
dc.subject Drug sensitivity en_US
dc.subject Murine en_US
dc.subject Ex vivo en_US
dc.subject Laboratory techniques and procedures en_US
dc.title A panel of Trypanosoma brucei strains tagged with blue and red-shifted luciferases for bioluminescent imaging in murine infection models en_US
dc.type Article-E en_US
dc.citation.issue 8 en_US
dc.citation.jtitle PLoS Neglected Tropical Diseases en_US
dc.citation.volume 8 en_US
dc.citation.pages e3054 en_US
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/25144573
dc.citation.jabbreviation PLoS Negl Trop Dis en_US


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