Figure 2.

The CXCR3 axis promotes T cell effector function during TIM-3 blockade. (A) Percent change in tumor volume from the start of PTX administration in mice treated with αTIM-3, αIFNγ and/or αCD8β as indicated. Merged data from two independent experiments; n=13–16 mice per group. (B) CD8⁺ T cells per mm² of tumor, quantified from whole tumor immunofluorescent images. Merged data from four independent experiments, n≥28 mice per group. (C) Representative histograms showing CXCR3 expression by tumor CD4⁺ and CD8⁺ T cells, as well as cDC1. (D) Mean fluorescence intensity (MFI) of surface CXCR3 on tumor T cells. Representative data from one of three independent experiments, n=9 mice. (E) Mice bearing PyMT tumors were treated with PTX and either αTIM-3 or the IgG_2a control, without (left) or with (right) αCXCR3. Merged data from two independent replicates; n≥14 mice per group. (F) Infiltration of CD8⁺ T cells (left) and CD4⁺ T cells (right) in the tumors from (E). Shown as a percent of CD45⁺ cells. (G) Percentage of CD8⁺ T cells expressing CD69, CD44, Ki67, or PD-1 after isolation from tumors treated with αTIM-3/PTX vs IgG_2a/PTX. Representative data from one of two independent experiments, n=10 mice per group. (G) Representative flow plots showing IFNγ expression by CD8⁺ T cells from PyMT tumors following ex vivo stimulation with PMA and ionomycin, isolated 12 days post PTX. (I) Quantitation of IFNγ expression by CD8⁺ T cells from (G). (J) Quantitation of IFNγ expression by restimulated CD8⁺ T cells isolated from mice treated with αTIM-3/PTX or IgG_2a/PTX,±AMG487. Merged data from two independent experiments; n≥8 mice per group. Significance in A and E determined by two-way analysis of variance. Significance in F, I, J determined by unpaired t-test. Shown as *p≤0.05, **p≤0.01. cDC1, type 1 conventional dendritic cell; IFNγ, interferon γ; ns, no significance; PD-1, programmed death-1; PMA, phorbol 12-myristate 13-acetate; PTX, paclitaxel; TIM-3, T cell immunoglobulin and mucin domain containing-3.