Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Apr 10;629(8010):211–218. doi: 10.1038/s41586-024-07300-8

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024, corrected publication 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Extended Data Fig. 3 — a, Schematic depicting engineering of truncated NGFR-only (tNGFR), TCF1/tNGFR- (TCF1_OE), and FOXO1/tNGFR- (FOXO1_OE) CAR T cells and magnetic isolation of tNGFR⁺ cells for downstream analyses. b–e, Phenotypic and functional analyses of CD19.BBζ CAR T cells at baseline and during repeat stimulation with Nalm6 cells. b,c, Flow cytometric analysis of CD62L and IL-7Rα (b) and TCF1 and LEF1 (c) from 1 representative donor (n = 4 donors). d, Cytokine secretion from CD19.BBζ cells after 1 or 4 stimulations with Nalm6 cells. Data show mean ± s.d. of 2–3 triplicate wells from 1 representative donor (n = 2 donors). e, Flow cytometric analysis of CD62L, IL-7Rα, and CD39 on tNGFR⁺ CD8⁺ CAR T cells prior to the first stimulation (Stim 0) and 7 days after the third stimulation (Stim 3). Data show mean ± s.e.m. of mean fluorescence intensity normalized to tNGFR levels from n = 2–3 donors. f–i, CAR T cell exhaustion model^7,23 whereby T cells express a high-affinity GD2-targeting CAR (HA.28ζ) that promotes antigen-independent tonic CAR signalling. f, Model schematic. g, Flow cytometric analysis of day 15 CD62L and IL-7Rα. Data show 1 representative donor (n = 5 donors). h, Cytotoxicity of day 15 HA.28ζ cells against 143B cells at a 1:8 E:T ratio. Data is normalized to t = 0 and show mean±s.d. of 3 triplicate wells from 1 representative donor (n = 3 donors). Statistics were performed at t = 96 h. i,j, Seahorse metabolic analyses on day 13 of culture (n = 2 donors). i, Ratio of OCR to ECAR of HA.28ζ cells. j, CD19.28ζ cell OCR (left), OCR to ECAR ratio (centre), and spare respiratory capacity (right). OCR line graph shows 1 representative donor. Bar graphs show mean ± s.d. of three representative time points within each donor. Statistical comparisons were performed using one-way ANOVA with Tukey’s test (h) or Dunnett’s test (i,j). E:T ratio, effector:target cell ratio. OCR, Oxygen consumption rate. ECAR, extracellular acidification rate.

Source Data