In vitro generation of CD8 Trm cells

The precise molecular mechanisms controlling tissue resident T cell (Trm cell) development remain unclear. In our studies, we have found that the vitamin A metabolite all-trans retinoic acid (atRA) synergises with TGFβ to promote Trm cells. We further show that polyclonal activation of naive splenic CD8 T cells in the presense of IL-2, IL-15, TGFβ + atRA leads to the development of CD69+CD103+ Trm cells. We have examined the transcriptome of CD8 T cells under IL-2/15, IL-2/15/atRA, IL-2/IL-15/TGFβ and IL-2/IL-15/TGFβ/atRA conditions. RNA isolation was conducted on day 7 from cells cultured in either IL-2 + IL-15, IL-2 + IL 15 + atRA, IL-2 + IL-15 + TGFβ, or IL-2 + IL-15 + TGFβ + atRA