1. Identification of a novel population of highly cytotoxic c‐Met‐expressing CD8+ T lymphocytes
Day 5 c‐Met+ or c‐Met− CD45.2+ Pmel‐1 CTLs treated with either PBS or HGF (30 ng/ml for 24 h) were transferred i.v. into congenic WT CD45.2+ recipients. Equal numbers of hgp10025–33‐pulsed CFSE‐labeled target CD45.1+ and unpulsed brilliant violet (BV)‐labeled CD45.1+ splenocytes were injected i.v. into the recipient CD45.2+ mice 5 h after the adoptive transfer of Pmel‐1 CTLs. Recipient CD45.2+ mice were sacrificed 24 h following transfer of target cells and control splenocytes, and the percentage of remaining target cells was quantified by FACS analysis. Representative dot plots of CFSE vs. BV profile of recovered CD45.1+ mouse splenocytes. Numbers indicate the percentage of cells of the high or null CFSE or BV phenotype. Graphical representation (scatterplot) of analyzed data pooled from two experiments (n = 3 mice/group). Controls referred to as non‐transferred. Percentage of in vivo cytotoxic activity was calculated as described in . Error bars show mean ± SEM, ***P < (unpaired, 2‐tailed Student's t‐test). Spleens were removed from each mouse at sacrifice, and content in GrB per gram of tissue was measured by ELISA. Error bars show mean ± SEM of 6 mice per group, ***P < (unpaired, 2‐tailed Student's t‐test). ELISpot assays were used to determine in individual mice the number of cells secreting IFN‐γ after ex vivo re‐stimulation of splenocytes with hgp10025–33 peptide for 24 h. Spots were counted with an ELISpot reader (Cellular Technology Ltd.) and expressed as spot‐forming cells per 1 × 106 splenocytes, after subtraction of control values (no peptide). Error bars show mean ± SEM, *P < 0.05 (unpaired, 2‐tailed Student's t‐test). A representative well image for each group is given on the left of the graph. Data are representative of two independent experiments (n = 3 mice/group).
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EMBO Rep, Volume: 18, Issue: 9, Pages: , First published: 27 July 2017, DOI: ( /embr )