Fig. 1 | Nature Communications

Fig. 1

From: CXCR1- or CXCR2-modified CAR T cells co-opt IL-8 for maximal antitumor efficacy in solid tumors

Fig. 1

IL-8 secretion is enhanced by IR in tumors. a Tumor chemokine changes after IR. U87 cells were radiated at 0 or 6 Gy and then seeded at 1 × 104/ml. Cells were harvested for RNA extraction 7 days later, followed by qPCR for chemokine profiling. The circle size corresponds to the mean values of relative gene expression (n = 4). b Kinetics of IL-8 secretion by tumor cells. U87 cells were treated with different doses of IR and seeded at 1 × 104/ml. Supernatants were harvested at 4, 24, 48, 96, or 168 hrs, and the IL-8 secretion was measured by ELISA (mean ± SD; n = 3; and the significance was determined by two-way ANOVA). c IL-8 production from multiple GBM lines. IL-8 expression was quantified by qPCR and ELISA, respectively, using established glioma lines (T98G and U118) and primary lines (pGBM-1129 and pGBM-1156) 7 days after IR. Tumor cells were irradiated at 0 or 6 Gy and plated at 1 × 104/ml, and cells and cell supernatants were harvested 7 days later for IL-8 production (means ± SEM; n = 3; and the Mann–Whitney U test was used). d NHA did not secrete IL-8 after IR. NHA and U87 cells were irradiated, and ELISA was carried out for IL-8 production 7 days after IR (means ± SEM; n = 3; and the Mann–Whitney U test was performed). eg IR elevates IL-8 expression in vivo. Design of the xenograft mouse model with local IR (e). Mice were injected intracranially with 5 × 104/mice of U87 cells. Eight days later, mice were locally irradiated with 0–3 fractionated doses daily at 3 Gy/day. Tumor samples were collected when mice were reaching the endpoint. Tumor  IL-8 gene (f) and protein (g) expression (means ± SEM; n = 5; by the Mann–Whitney U test) measured by qPCR or IHC. All the experiments were repeated at least three times. h Schema: IR enhances the expression of IL-8 by tumors that can be co-opted by IL-8 receptor-expressing CAR T cells to increase trafficking of CAR T cells to tumors. *p < 0.05, **p < 0.01, ***p < 0.001

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