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Learn MoreWe sought to describe in detail the consequences of MAIT cell activation using a transcriptomic approach to define the basic transcriptome of a MAIT cell in both humans and mice and to determine how this is modulated by activation. Fresh human peripheral blood cells were obtained from three donors. These were cultured for 6 hours with (stimulated) or without (unstimulated) 10 nM 5-OP-RU, magnetically enriched on MR1-tetramer+ cells, and flow-sorted for RNA sequencing of live CD3+TCR Valpha7.2+ MR1-5-OP-RU tetramer+ MAIT cells, and of unstimulated nave live CD8+CD45RA+ T cells as a comparator cell type. For the murine samples we included within the same sequencing experiment live pulmonary CD3+45.2+19-MR1-5-OP-RU tetramer+ MAIT cells which were magnetically enriched and flow-sorted from the lungs of mice 7 days after infection with 1x104 CFU L. longbeachae (acute), or at least 12 weeks post infection (resolution) or 7 days after a second intranasal infection with 2x104 CFU L. longbeachae in mice that had recovered from infection 12 weeks previously (reinfection). Live CD3+CD45.2+CD19-CD8+CD44-CD62L+ nave T cells from uninfected mice were used as a comparator cell type. SOURCE: Paul Klenerman (paul.klenerman@ndm.ox.ac.uk) - NDM Oxford
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