Figure S6 . " width="100%" height="100%">
Journal: Cell Reports Medicine
Article Title: Development of a chimeric cytokine receptor that captures IL-6 and enhances the antitumor response of CAR-T cells
doi: 10.1016/j.xcrm.2024.101526
Figure Lengend Snippet: Functional and phenotypic properties of G6/7R-M452L anti-mesothelin CAR-T cells in a solid tumor model (A) NSG mice subcutaneously inoculated with AsPC1 were infused with T cells engineered with anti-mesothelin CAR alone or anti-mesothelin CAR and G6/7R-M452L. CAR-T cells were cotransduced with the luciferase gene. (B) Monitoring of the subcutaneous tumor volume ( n = 7 mice for each). (C) Progression-free survival of the treated mice ( n = 7 mice, log rank test). (D) Frequency of human T cells in the peripheral blood at the indicated time points ( n = 7 mice, unpaired two-tailed Student’s t test). (E) The total flux of the luciferase activity was analyzed by in vivo bioluminescent imaging ( n = 7 mice, unpaired two-tailed Student’s t test of the log-transformed values). In (A)–(E), the data are a composite of two independent experiments. ∗ p < 0.05, ∗∗ p < 0.01. (F and G) The subcutaneous tumor was harvested 6–9 weeks following anti-mesothelin CAR-T cell infusion in the AsPC-1 tumor model shown in (A). (F) Percentage of CD8 + CAR-T cells expressing exhaustion markers (PD-1, LAG-3, and TIM-3) ( n = 6 mice for the CAR alone group and n = 8 mice for the CAR+G6/7R-M452L group). Error bars denote SD. (G) Phospho-STAT3 and STAT5 levels in the CD8 + CAR-T cell population were quantified by flow cytometry ( n = 8, unpaired two-tailed Student’s t test). (H) AsPC-1-bearing NSG mice were infused with G6/7R or G6/7R-M452L anti-mesothelin CAR-T cells. CAR-T cells persisting in the spleen were analyzed for Ki-67 expression ( n = 8 mice, unpaired two-tailed Student’s t test). (I) Anti-mesothelin G6/7R-M452L CAR-T cells persisting after regression of AsPC-1 were collected from spleen and transplanted into tumor-free NSG mice ( n = 9 mice). (J) The frequency of human T cells in the peripheral blood of mice with secondary transplantation was analyzed by flow cytometry ( n = 9 mice). (K) Total flux of the luciferase expressed in CAR-T cells was serially monitored by in vivo imaging. For comparison, luciferase-transduced Jurkat cells were transplanted into NSG mice and monitored using the same protocol ( n = 9 mice for the CAR-T cell group, and n = 8 mice for the Jurkat group). In (D), (E), (G), (H), (J), and (K), horizontal lines denote mean values. See also Figure S6 .
Article Snippet: PE-Cy7 anti-human CD279 (PD-1) , BD Pharmingen , Cat#561272; RRID: AB_10611585.
Techniques: Functional Assay, Luciferase, Two Tailed Test, Activity Assay, In Vivo, Imaging, Transformation Assay, Expressing, Flow Cytometry, Transplantation Assay, In Vivo Imaging, Comparison
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