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

letter

. 2021 Jun 4;11(6):108. doi: 10.1038/s41408-021-00499-z

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

© The Author(s) 2021

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 1 — A PD-L1 expression on Nalm-16 cells was analyzed 24 h after stimulation with IFN-γ and TNF-α. B Primary ALL blasts from 31 different pediatric BCP-ALL patients were stimulated with IFN-γ and TNF-α for 24 h. Pie chart shows percentage of PD-L1-inducible vs. PD-L1-non-inducible samples (left panel). Surface PD-L1 expression in inducible patients (n = 16) is shown in the right panel (fold-change of stimulated sample vs. unstimulated sample). C Conventional second-generation CAR T cells were co-cultured with CD19⁺ and CD19⁺/PD-L1⁺ Daudi cells. Percentage of intracellular IFN-γ⁺ in CAR T cells was analyzed 24 h later. D 19_BB_3z T cells were co-cultured with Daudi cells for 24 h and co-culture supernatant was analyzed for concentration of IFN-γ, IL-2 and TNF-α in a flow-cytometry-based assay. E Schematic illustration of CAR constructs and negative control (19t). F 19_3z_PD-1_28 CAR T cells were co-cultured with CD19^-/PD-L1^-, CD19^-/PD-L1⁺ and CD19⁺/PD-L1⁺ target cells (transduced K562 cells). Intracellular cytokine stain was performed 24 h later and showed no unspecific activation of the PD-1-CD28 fusion protein. G Cytotoxicity of conventional second-generation CAR T cells (19_BB_3z) and second-generation CAR T cells with fusion protein (19_BB_3z_PD-1_28) was analyzed 48 h after co-culture with CD19⁺/PD-L1⁺ K562 cells. H CAR T cells were co-cultured with CD19⁺ and CD19⁺/PD-L1⁺ Daudi cells. Intracellular stain for IFN-γ was performed 24 h later. I CAR T cells were co-cultured with CD19⁺/PD-L1⁺ target cells and concentration of IL-2 and TNF-α was analyzed in the supernatant 24 h after start of co-culture. J Schematic illustration of the second-generation anti-CD22 CAR and the version with fusion protein used in this study. K Intracellular cytokine stains of anti-CD22 CAR T cells 24 h after co-culture with Daudi cells showed increased cytokine release of PD-1-CD28 CAR T cells. L Activation markers 4-1BB, CD25 and CD69 were increased in PD-1-CD28 CAR T cells over conventional anti-CD22 CAR T cells 24 h after co-culture with Nalm-6 cells. N ≥ 3 individual donors (C, D, F, G, H and I). N ≥ 2 individual donors (K and L). Statistical significance was calculated using t-test. UT untransduced T cells, E:T ratioeffector to target ratio, FC fold-change, BCP-ALL B-cell precursor ALL, n.s. not significant, MFI geometric mean fluorescent intensity.