Figure S1.
Wound healing scratch assay. (A) Schematic of three-cell-type system with HLA-A2+ fibroblast cells (MRC-5) presenting influenza peptide to influenza-specific CD8 T cells, and HLA-A2− epithelial cells (HaCaT cells, keratinocytes). Visualization was created with BioRender using a graphical element from Fig. 1 B. (B) Expression of HLA-A2 in HaCaT versus MRC-5 cell lines. (C) Depiction of proliferation mask of HaCaT cells in a coculture with MRC-5 cells seeded in a 30:70 HaCaT:MRC-5 cell ratio pulsed with 0 (left) or 100 ng/ml (right) influenza peptide and cocultured with influenza-specific T cells after 0 (top) or 5 (bottom) h. Raw images are depicted in Fig. 1 A. Scale bars = 400 µm; enhanced for improved visibility. (D) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with varying amounts of influenza peptide for 1 h. Cells were cultured in the presence of influenza-specific T cells and cell-free SN was tested in a wound healing assay with HaCaT cells; two independent assay results (n = 3). (E) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with 100 ng/ml influenza peptide for 1 h. Cells were cultured in the presence or absence of influenza-specific T cells o/n. Cells were then either washed 3× and cultured o/n with fresh medium in the absence of influenza-specific T cells (preactivation) or left untouched (continuous activation). After an additional 24 h incubation period, cell-free SN was harvested and tested in a wound healing assay with HaCaT cells (n = 4). (F) Measurement of intracellular TNF and IFN-γ in influenza-specific T cells used in the combined proliferation and killing assay; representative gating. (G and H) Individual, unnormalized experiments of AREG (G) or TGFα (H) ELISA from Fig. 1 D. (I) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with 0 or 100 ng/ml influenza peptide for 1 h. Cells were cultured in the presence of influenza-specific T cells and cell-free SN was tested in a wound healing assay with HaCaT cells in the presence of aIFNγ, Cetuximab, aIFNγ, and Cetuximab or IgG control (n = 4).

Wound healing scratch assay. (A) Schematic of three-cell-type system with HLA-A2+ fibroblast cells (MRC-5) presenting influenza peptide to influenza-specific CD8 T cells, and HLA-A2− epithelial cells (HaCaT cells, keratinocytes). Visualization was created with BioRender using a graphical element from Fig. 1 B. (B) Expression of HLA-A2 in HaCaT versus MRC-5 cell lines. (C) Depiction of proliferation mask of HaCaT cells in a coculture with MRC-5 cells seeded in a 30:70 HaCaT:MRC-5 cell ratio pulsed with 0 (left) or 100 ng/ml (right) influenza peptide and cocultured with influenza-specific T cells after 0 (top) or 5 (bottom) h. Raw images are depicted in Fig. 1 A. Scale bars = 400 µm; enhanced for improved visibility. (D) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with varying amounts of influenza peptide for 1 h. Cells were cultured in the presence of influenza-specific T cells and cell-free SN was tested in a wound healing assay with HaCaT cells; two independent assay results (n = 3). (E) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with 100 ng/ml influenza peptide for 1 h. Cells were cultured in the presence or absence of influenza-specific T cells o/n. Cells were then either washed 3× and cultured o/n with fresh medium in the absence of influenza-specific T cells (preactivation) or left untouched (continuous activation). After an additional 24 h incubation period, cell-free SN was harvested and tested in a wound healing assay with HaCaT cells (n = 4). (F) Measurement of intracellular TNF and IFN-γ in influenza-specific T cells used in the combined proliferation and killing assay; representative gating. (G and H) Individual, unnormalized experiments of AREG (G) or TGFα (H) ELISA from Fig. 1 D. (I) MRC-5 and HaCaT cells were seeded in a 30:70 ratio and pulsed with 0 or 100 ng/ml influenza peptide for 1 h. Cells were cultured in the presence of influenza-specific T cells and cell-free SN was tested in a wound healing assay with HaCaT cells in the presence of aIFNγ, Cetuximab, aIFNγ, and Cetuximab or IgG control (n = 4).

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