Dataset Information

Development of Trypanosoma brucei EATRO 1125 in the host bloodstream

ABSTRACT: Trypanosoma brucei EATRO 1125 parasites were grown in adult MF1 mice with parasites harvested on day 3 post-infection ('ascend/slender') or on day 6 post-infection ('peak/stumpy'). Overall design: Three replicates each of two conditions (ascending/slender, peak/stumpy) were analysed. 'Peak' samples were each derived from infections in three individual mice; 'ascend' samples were derived from 2 individual mice infections (replicate 1 and 2) and from a pool of four infections (replicate 3), to generate sufficient material for analysis. All samples were analysed for their cell cycle status by scoring the number of kinetoplasts and nuclei in parasites and were assayed for PAD1 expression, a marker of stumpy formation.

INSTRUMENT(S): Illumina HiSeq 2000 (Trypanosoma brucei brucei)

SUBMITTER: Alasdair Ivens

PROVIDER: GSE114810 | GEO | 2018-10-01

REPOSITORIES: GEO

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			Action	DRS
	GSE114810_Tb_rawcounts2.txt.gz	Txt

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Publications

A gene expression comparison of Trypanosoma brucei and Trypanosoma congolense in the bloodstream of the mammalian host reveals species-specific adaptations to density-dependent development.

Silvester Eleanor E Ivens Alasdair A Matthews Keith R KR

PLoS neglected tropical diseases 20181011 10

In the bloodstream of mammalian hosts Trypanosoma brucei undergoes well-characterised density-dependent growth control and developmental adaptation for transmission. This involves the differentiation from proliferative, morphologically 'slender' forms to quiescent 'stumpy' forms that preferentially infect the tsetse fly vector. Another important livestock trypanosome, Trypanosoma congolense, also undergoes density-dependent cell-cycle arrest although this is not linked to obvious morphological t ...[more]

PMID: 30307943

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Project description:Human African trypanosomiasis is caused by infection with parasites of the Trypanosoma brucei species complex, and threatens over 70 million people in sub-Saharan Africa. Development of new drugs is hampered by the limitations of current rodent models, particularly for stage II infections, which occur once parasites have accessed the CNS. Bioluminescence imaging of pathogens expressing firefly luciferase (emission maximum 562 nm) has been adopted in a number of in vivo models of disease to monitor dissemination, drug-treatment and the role of immune responses. However, lack of sensitivity in detecting deep tissue bioluminescence at wavelengths below 600 nm has restricted the wide-spread use of in vivo imaging to investigate infections with T. brucei and other trypanosomatids.Here, we report a system that allows the detection of fewer than 100 bioluminescent T. brucei parasites in a murine model. As a reporter, we used a codon-optimised red-shifted Photinus pyralis luciferase (PpyRE9H) with a peak emission of 617 nm. Maximal expression was obtained following targeted integration of the gene, flanked by an upstream 5'-variant surface glycoprotein untranslated region (UTR) and a downstream 3'-tubulin UTR, into a T. brucei ribosomal DNA locus. Expression was stable in the absence of selective drug for at least 3 months and was not associated with detectable phenotypic changes. Parasite dissemination and drug efficacy could be monitored in real time, and brain infections were readily detectable. The level of sensitivity in vivo was significantly greater than achievable with a yellow firefly luciferase reporter.The optimised bioluminescent reporter line described here will significantly enhance the application of in vivo imaging to study stage II African trypanosomiasis in murine models. The greatly increased sensitivity provides a new framework for investigating host-parasite relationships, particularly in the context of CNS infections. It should be ideally suited to drug evaluation programmes.