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Conclusion:
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In line with the observations of VAC069A, infection led to widespread recruitment of innate-like and adaptive T cells out of circulation during this study. After drug treatment (and clearance of parasites) T cells returned to circulation at near-normal levels within six days. Atthis time-point, 10-20% of the T cell compartment was activated (CD38hi Bcl2lo) in first infection (v11 and v21). In contrast, the three rechallenged volunteers (v02, v05 and v07) exhibited reduced levels of activation (5-9%). Importantly, a direct comparison between VAC069A and VAC069B showed that T cell activation was reduced within these three volunteers by 25-50% in second (compared to first) infection. The lower levels of activation were particularly evident amongst conventional CD4 T cells and to a lesser extent innate-like T cells (e.g. gamma delta and MAIT cells). These data are analogous to our findings in a rechallenge model of falciparum malaria (flowrepository.org experiment number FR-FCM-Z464), which show that T cell activation is attenuated upon reinfection despite an identical pathogen load.
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Comments:
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Staining and acquisition was performed at the MRC Weatherall Institute for Molecular Medicine in Oxford, UK, in collaboration with Michalina Mazurczyk and Giorgio Napolitani. CyTOF was used to resolve changes in the peripheral T cell pool through time in five volunteers (named v02, v05, v07, v11, v21). Demographic information of this cohort is shown in a spreadsheet attached to this experiment. Each volunteer was sampled at four timepoints: C-1 (one day before infection), DoD (day of diagnosis, immediately before treatment), ep6 (six days post-treatment) and C56, 56 days post challenge (~40 days post treatment). The filename contains both the timepoint and individual from whom the sample
was taken. FCS files were generated, followed by bead normalisation and debarcoding using CyTOF software (version 6.7, Fluidigm). All following analyses were carried out in R. T cells were gated by first performing flowSOM clustering to export CD45+CD3+ clusters from whole blood followed by singlet gating using openCyto yielding a Ir191+Ir193+Ce140- population. These singlet T cells were taken forward for CATALYST/flowSOM/diffcyt workflow described in Nowicka et al. (2019 F1000 Research). FlowSOM metaclustering was used to classify T cells into unique subsets. This repository contains fcs files that have been pregated on CD45+CD3+ singlet T cells. Ungated whole blood will shortly be uploaded to a separate experiment. Should this be of interest to you ahead of publication, please get in touch and we will be happy to share.
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