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Schematic of the experimental design. CD45.1 donor T cells specific to OVA (OT-I) or gB (gBT-I) were adoptively transferred into CD45.2 hosts with and without intranasal co-administration of polyI:C and OVA or gB peptides to generate high and low levels of CD8+TRM-like cells specific to OVA (CD45.1+tumor-specific OT-I TRM-like cells: t-TRMcells) or gB (CD45.1+bystander gBT-I TRM-like cells: by-TRM) compared with no TRMcontrols. Lung tumor cells generated from aKrasG12D/+;p53flox/floxmouse were engineered to express OVA (KP-OVA) and injected 2 weeks after transfer of CD45.1 cells to generate experimental groups with pre-existing TRM-like cells specific to the tumor OVA antigen (t-TRM) or pre-existing TRM-like cells reactive to gB (by-TRM). Lungs were collected either prior to tumor cell injection (n = 3/group) or 8 weeks after tumor cell injection (n = 6/group). Mice were injected with anti-CD3-AF532 5 min before collection to label circulating T cells.

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