Establishment of a panel of reference Trypanosoma evansi and Trypanosoma equiperdum strains for drug screening

Gillingwater, Kirsten; Büscher, Philippe; Brun, Reto

doi:10.1016/j.vetpar.2007.05.020

Cited by 25 publications

(17 citation statements)

References 28 publications

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“…gambiense than T.b. brucei to suramin has been described previously, but an even higher resistance has been described in T. evansi [53], [54]. T.b.…”

Section: Discussionmentioning

confidence: 78%

“…Dophanil powder (Docpharma, Hoeilaart, Belgium), containing 455 mg diminazene diaceturate and 555 mg antipyrine per gram, was prepared as a 10 mg ml −1 diminazene diaceturate solution in DMSO. A method to measure the IC 50 values of compounds in 96-well plates was performed as described elsewhere [53]. Threefold drug dilutions in duplicate were made in HMI-9 medium to allow testing in final drug concentrations ranging from 100 to 0.14 µg ml −1 for eflornithine and hygromycin, from 50 to 0.07 µg ml −1 for nifurtimox, from 10 to 0.014 µg ml −1 suramin and from 500 to 0.7 ng ml −1 for diminazene diaceturate, melarsoprol and pentamidine with 5×10 3 cells ml −1 in a total volume of 200 µl.…”

Section: Methodsmentioning

confidence: 99%

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A Panel of Trypanosoma brucei Strains Tagged with Blue and Red-Shifted Luciferases for Bioluminescent Imaging in Murine Infection Models

et al. 2014

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BackgroundGenetic 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 findingsWe 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/SignificanceWe 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.

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“…gambiense than T.b. brucei to suramin has been described previously, but an even higher resistance has been described in T. evansi [53], [54]. T.b.…”

Section: Discussionmentioning

confidence: 78%

Section: Methodsmentioning

confidence: 99%

A Panel of Trypanosoma brucei Strains Tagged with Blue and Red-Shifted Luciferases for Bioluminescent Imaging in Murine Infection Models

et al. 2014

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“…We therefore supplemented adenosine, which is mainly taken up via nucleoside transporters P1 and P2 (Carter and Fairlamb, 1993). T. equiperdum OVI has a substantially reduced sensitivity for diamidines like the drug diminazene, which is taken up via P2 (Barrett et al, 1995;Gillingwater et al, 2007). To definitively compensate for the altered nucleoside transporter we added hypoxanthine, which is mainly taken up via the distinct nucleobase transporters, H1 and H4 (Natto et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

In vitro production of Trypanosoma equiperdum antigen and its evaluation for use in serodiagnosis of dourine

Bassarak

Moser

Menge

2016

Veterinary Parasitology

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“…However, this has not been established as an effective treatment strategy for dourine-infected horses. Moreover, diminazene- and melarsomine-resistant T. equiperdum strains have already been reported (Gillingwater et al., 2007). Thus, an in vitro drug screening assay for T. equiperdum is strongly needed for efforts to seek novel trypanocidal compounds both for evaluating drug sensitivity and for establishing an effective treatment strategy for dourine.…”

Section: Introductionmentioning

confidence: 95%

“…Dourine is listed as an internationally important animal disease by the World Organization for Animal Health (OIE). Evidence from in vitro drug sensitivity assays using laboratory strains of T. equiperdum indicates that suramin, diminazene, quinapyramine and Cymelarsan ® (melarsomine) are effective against T. equiperdum (Zhang et al., 1991, Brun and Lun, 1994, Gillingwater et al., 2007). Melarsomine was found to be effective against the T. equiperdum OVI strain in mice, and the T. equiperdum Dodola strain in mice and horses (Hagos et al., 2010, Habte et al., 2015).…”

Section: Introductionmentioning

confidence: 99%

The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

Suganuma

Yamasaki

Molefe

et al. 2017

International Journal for Parasitology: Drugs and Drug Resistan

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Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE), no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T. equiperdum strain was recently isolated from the genital organ of a stallion that showed typical symptoms of dourine. In the present study, we adapted T. equiperdum IVM-t1 from soft agarose media to HMI-9 liquid media to develop a drug screening assay for T. equiperdum. An intracellular ATP-based luciferase assay using CellTiter-Glo reagent and an intracellular dehydrogenase activity-based colorimetric assay using WTS-8 tetrazolium salt (CCK-8 reagent) were used in order to examine the trypanocidal effects of each compound. In addition, the IC50 values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine) were evaluated and compared using established assays. The IC50 values of these reference compounds corresponded well to previous studies involving other strains of T. equiperdum. The luciferase assay would be suitable for the mass screening of chemical libraries against T. equiperdum because it allows for the simple and rapid-evaluation of the trypanocidal activities of test compounds, while a simple, inexpensive colorimetric assay will be applicable in developing countries for the evaluation of the drug sensitivity of epidemic trypanosome strains.

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Establishment of a panel of reference Trypanosoma evansi and Trypanosoma equiperdum strains for drug screening

Cited by 25 publications

References 28 publications

A Panel of Trypanosoma brucei Strains Tagged with Blue and Red-Shifted Luciferases for Bioluminescent Imaging in Murine Infection Models

A Panel of Trypanosoma brucei Strains Tagged with Blue and Red-Shifted Luciferases for Bioluminescent Imaging in Murine Infection Models

In vitro production of Trypanosoma equiperdum antigen and its evaluation for use in serodiagnosis of dourine

The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

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