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. 2022 Apr 20;10(4):e004218. doi: 10.1136/jitc-2021-004218

Figure 3.

Figure 3

BEMPEG+NKTR-262 induces a greater expansion of active CD8+ T cells than BEMPEG+RT. (A) We determined immune phenotypes from PBL 7 days post-therapy. These data are presented as Spearman correlations of PBL phenotypes with tumor size, across individuals and treatment groups. Significant correlations after FDR correction are indicated. (B) Representative flow cytometry plots of CD45+, CD4+, and CD8+ gates. (C) Representative flow cytometry plots demonstrating CD8+ cell expression of CD62L, PD-1, GzmA, ICOS, Ki-67, and AH1-A5 after BEMPEG+RT (blue) or BEMPEG+NKTR-262 (red). (D) PBL immune phenotypes determined by flow cytometry. Box and whisker plots represent the min and max (whiskers), the quartiles (box) and median (line). Each point represents an individual mouse. N=10–20, from two independent experiments, except AH1-A5, which is one of two representative experiments N=4–8 per experiment. One-way ANOVA with Šídáks multiple comparisons test. (E) (Left) Representative flow cytometry plot showing AH1-A5+ and GzmA+ CD8+ T cells. (right) Granzyme A (GzmA), proliferation (Ki-67), and CD62L expression on tumor specific (AH1-A5+, filled circle) or not (AH1-A5-, open circle). Only mice that had more than 70 AH1-A5+ cells were analyzed for AH1-A5+ phenotypes. Data from one of two representative experiments. For comparisons between AH1-A5+/- within one treatment group, Student’s t-test. For comparisons among treatment groups, one-way ANOVA with Šídáks multiple comparisons test. *P<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; FDR, false discovery rate; PBL, peripheral blood; RT, radiation therapy.