Fig. 1: B7-H3.CAR-T cells target B7-H3+ NSCLC cell lines and organoid in vitro. | Nature Communications

Fig. 1: B7-H3.CAR-T cells target B7-H3+ NSCLC cell lines and organoid in vitro.

From: Targeting brain lesions of non-small cell lung cancer by enhancing CCL2-mediated CAR-T cell migration

Fig. 1

a Representative micrographs showing B7-H3 expression in cryosections of normal lung and NSCLC assessed by staining with the 376.96 mAb at the final concentration of 1 μg/mL. Scale bars, 200 μm. b B7-H3 expression score summary of the immunochemistry results in normal lung and NSCLC showing in (a), n = 3 for normal lung, n = 5 for adenocarcinoma, n = 30 for squamous cell carcinoma. Data are presented as mean values ± SD. c Representative flow cytometry plots showing B7-H3 expression in NSCLC cell lines, stained with the 376.96 mAb antibody. d, e NSCLC cell lines labeled with GFP were cocultured with CD19.28, B7-H3.28, or B7-H3.BB CAR-T cells at the T cell to tumor cell ratio of 1:5. On day 5, NSCLC cells (GFP+) and CAR-T cells (CD3+) were enumerated by flow cytometry. Representative flow-cytometry plots (d) and quantification of residual tumor cells (e) are illustrated. Data are presented as mean values + SD. f, g Summary of IFNγ (f) and IL2 (g) released by CAR-T cells in the culture supernatant after 24 h of coculture with the indicated cell lines as measured by ELISA, n = 4 independent experiments using CAR-T cells generated from four different donors in (d–g). Data are presented as mean values + SD. h Representative flow plots showing B7-H3 expression in lung cancer organoid LU6438B, which was stained with the 376.96 mAb antibody. i, j Organoid LU6438B was cocultured with CD19.28 or B7-H3.28 CAR-T cells at the T cell to organoid cell ratio of 1:2 and 1:5. On day 4, organoid cells (CD45–) and CAR-T cells (CD45+) were identified by flow cytometry. Representative flow-cytometry plots (i) and quantification of residual organoid cells (j) are illustrated. Data are presented as mean values. k, l Summary of IFNγ (k) and IL2 (l) released by CAR-T cells in the supernatant after 24 h of coculture with organoids as measured by ELISA. Data are presented as mean values. N = 2 independent experiments using CAR-T cells generated from two different donors. Source data for (b, e, f, g, j, k, l) are provided as a Source Data file.

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