Journal: Advanced Science
Article Title: GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
doi: 10.1002/advs.202408086
Figure Lengend Snippet: Anti‐GRP78 PE38 immunotoxin exerts potent antitumor effects against colorectal cancer and primary melanoma. a) Schematic of HT29 colorectal cancer model treated with PBS, Nb C5 (0.08 mg kg −1 ), or C5‐PE38 (0.3 mg kg −1 ) as indicated. BALB/c nude mice were injected subcutaneously with 5 × 10 6 HT29 tumor cells. b–d) Tumor growth and Kaplan‐Meier survival analysis of HT29 tumor‐bearing mice (mean ± SEM, n = 6–7 mice per group). Mice were euthanatized as an endpoint when the tumor volume reached 1500 mm 3 for survival analysis. e) Schematic of the B16F10 melanoma model treated with PBS, Nb C5, or C5‐PE38 as indicated. C57BL/6 mice were injected intradermally with 5 × 10 5 B16F10 tumor cells. f,g) Tumor growth of B16F10 tumor‐bearing mice following treatment (mean ± SEM, n = 4–5 mice per group). h) Tumor weight from B16F10 tumor‐bearing mice on day 9 post‐treatment (mean ± SEM, n = 4–5 mice per group). i) Representative images and quantification of Ki67 and TUNEL staining in tumor sections from the B16F10 tumor model following different treatments (mean ± SEM, n = 3 mice per group). H&E staining was performed for histological analysis. Scale bars, 100 µm (H&E), 50 µm (Ki67 and TUNEL). j) Representative immunofluorescence images and quantification of CALR staining in tumor sections from the B16F10 tumor model (mean ± SEM, n = 4 mice per group). Scale bars, 50 µm. Ki67‐positive, TUNEL‐positive, and CALR‐positive cell frequencies were quantified using ImageJ software. Statistical significance was assessed by two‐way ANOVA with Tukey's multiple‐comparisons (b, f), log‐rank Mantel‐Cox test (d), one‐way ANOVA with Tukey's multiple‐comparisons (h, i), and unpaired Student's t ‐test (j). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.
Article Snippet: Human colorectal cancer cell lines (HT29, SW620), murine colorectal cancer cell lines (CT26, MC38), murine melanoma cell lines (B16, B16F1, B16F10), murine breast cancer cell line (4T1), murine dendritic cell line (DC2.4), and human embryonic kidney cell (HEK293) were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China) or ATCC (USA).
Techniques: Injection, TUNEL Assay, Staining, Immunofluorescence, Software