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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Nat Biotechnol. 2015 Nov 2;33(12):1287–1292. doi: 10.1038/nbt.3386

Figure 1. JEDI T-cells specifically kill EGFP-expressing cells and can be used to study pathogen clearance and model autoimmune disease.

Figure 1

(a) Schematic of the methodology used to make the JEDI mice. (b) Splenocytes from JEDI mice were stained with CD3e, CD4, CD8a antibody, and an H-2Kd-GFP200-208 pentamer to measure the frequency of EGFP-specific T-cells. (c) Mice were injected with a lentivirus expressing EGFP (LV.EGFP) and 2 days later JEDI or control (Ctrl) CD8+ T-cells were injected. Flow cytometry was performed to measure EGFP-expressing cells in the spleen after 5 days. Shown is a representative dotplot from two separate experiments. Graph presents mean±s.d. of the percentage of EGFP+ cells among live splenocytes (n= 7 mice/group). **P<0.01 vs Control-treated. (d) Fluorescent microscopy analysis of the spleen of mice described in (c). Representative images are shown. White bar represents 100 μm. (e) Mice were injected with LV expressing EYFP (LV.EYFP) and 2 days later JEDI or control CD8+ T-cells were injected. Flow cytometry was performed to measure EYFP-expressing cells in the spleen after 5 days. Shown is a representative dotplot from two experiments (left). Graph presents the mean±s.d. of EYFP+ cells among live splenocytes (n=3 mice/group). (f) Bone marrow cells were transduced with LV.EYFP or LV.EGFP and differentiated into dendritic cells (BMDCs). Isolated JEDI or B10D2 CD8+ T-cells were stained with Violet Cell Proliferation dye and added to BMDCs. EGFP-specific cells were identified by H-2Kd-GFP200-208 pentamer staining. Representative flow cytometry plots are shown (n=3 wells/group). (g) MIP-EGFP mice were injected with 1×106 JEDI or control CD8+ T-cells, and vaccinated with EGFP. Fluorescent microscopy analysis of the pancreas was performed 6 days after T-cell transfer. Representative images are shown (n=4 mice/group). Arrows point to CD8+ cells in the tissue. White bar represents 100 μm. (h) Flow cytometry of H-2Kd-GFP200-208 pentamer positive CD8+ T-cells in the brachial and pancreatic lymph nodes of MIP-EGFP mice injected with JEDI T-cells as in (g). (i) MIP-EGFP mice were injected with 2×106 Violet Cell Proliferation dye-labeled JEDI or control CD8+ T-cells, and 6 days later proliferation was measured in the CD8+ T-cells in the pancreatic lymph nodes. EGFP-specific T-cells were identified by H-2Kd-GFP200-208 pentamer staining. Representative histogram is shown (n=2 mice/group). (j) Blood glucose measurements at the indicated times of mice described in (g) (n=4 mice/group).