Journal: Journal for immunotherapy of cancer
Article Title: Vaccination generates functional progenitor tumor-specific CD8 T cells and long-term tumor control.
doi: 10.1136/jitc-2024-009129
Figure Lengend Snippet: Figure 2 TST activate and proliferate in mice with early and late lesions. (A) Scheme: CFSE-labeled TCRTAG were adoptively transferred into C57BL/6 (B6; naive, gray), early (blue) and late (orange) AST;Cre-ERT2 mice and analyzed 60 hours later. (B) Top, flow analysis of TCRTAG CFSE dilution in the spleens, liver-draining lymph nodes (ldLN) and livers of early, late, and naive mice. This and all subsequent flow plots are gated on live CD8+Thy1.1+ cells (representative gating is shown in online supplemental figure 1A). Data are concatenated from three biological replicates and representative of three independent experiments. Bottom, percentage of TCRTAG in each cell division for spleen, ldLN, and liver. Each symbol represents an individual mouse with n=6–8/ group. *p<0.05, **p<0.01, ****p<0.0001 (two-way ANOVA followed by post hoc Šídák test). Biological replicates from two independent experiments were combined. (C) Left, PD1 expression directly ex vivo, and TNFα/IFNγ production after ex vivo TAG peptide stimulation, plotted against CFSE dilution. Naive T cells were assessed only from the spleen and shown as dark gray histograms or dots, naive T cells from the spleen are shown again in light gray in the ldLN and spleen plots for reference. Data are concatenated from three biological replicates and representative of two independent experiments for spleens and livers, and one experiment for ldLN. Right, percentage of TCRTAG positive for PD1, TNFα and IFNγ. Gates for this and subsequent cytokine production figures were set based on no peptide stimulation controls. Each symbol represents an individual mouse with n=7–8/ group for spleen and liver and n=3/group for ldLN. ns=not significant, *p<0.05, **p<0.01, ****p<0.0001 (unpaired Student’s t- test). (D) Number of TCRTAG in spleens, ldLN, and livers of early and late mice. Each symbol represents an individual mouse with n=10–13/group combined from three independent experiments for spleens and livers, and n=6 combined from two independent experiments for ldLN. *p<0.05, ***p<0.001, ****p<0.0001 (unpaired Student’s t-test). ANOVA, analysis of variance; CFSE, carboxy fluorescein succinimidyl ester.
Article Snippet: The decreased TST proliferation in mice Table 2 Flow cytometry cell dyes Dye name Source Identifier CFSE Tonbo Cat# 13- 0850 Ghost Dye Red 780 Tonbo Cat# 13- 0865 Cell- surface and intracellular cytokine staining.
Techniques: Labeling, Expressing, Ex Vivo
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