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 8;12(4):e008429. doi: 10.1136/jitc-2023-008429

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

© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

PMC Copyright notice

PD-1 inhibition promotes the memory phenotype and reduces the exhaustion of BCMA CAR-T cells. (A) Schematic diagram of the CAR-T cell cytotoxicity assay after long-term repetitive antigen stimulation. (B) CAR expression was determined by flow cytometry in PD-1^KD BCMA CAR-T cells (upper) and BCMA CAR-T cells (lower) after fourth repetitive stimulation (d13). Representative (B) and summarized (E) data from three representative healthy donors are shown. (C) Annexin V and PI-staining of PD-1^KD BCMA CAR+T and BCMA CAR+T cells after four repetitive stimulations (d13). Representative (C) and summarized (F) data from three representative healthy donors are shown. (D) Representative differentiation statuses of PD-1^KD BCMA CAR-T cells (upper) and BCMA CAR-T cells (lower) cells were determined by flow cytometry using the surface markers CD45RA and CD62L after four repetitive stimulations (d13). (G, H) Ratios of central memory (Tcm; CD45RA−CD62L+) and effector memory (Tem; CD45RA−CD62L−) cells were compared for PD-1^KD BCMA CAR-T and BCMA CAR-T cells. (I) GSEA of central memory genes. The central memory genes are shown in online supplemental table 3. The genes from the left to the right of the rank-ordered list are enriched in PD-1^KD BCMA CAR-T cells and BCMA CAR-T cells. (J) GSEA of effector versus exhausted up genes. Effector versus exhausted upgenes are shown in online supplemental table 4. The genes from the left to the right of the rank-ordered list are enriched in PD-1^KD BCMA CAR-T cells and BCMA CAR-T cells. (K, L) Volcano plots of genes differentially expressed between PD-1^KD BCMA CAR+T cells and BCMA CAR+T cells sorted after one antigen stimulation (d1) or after four repetitive stimulations (d13). Significantly differentially expressed genes associated with the memory phenotype are highlighted with an orange circle. (M) Heatmap of normalized RNA-seq reads for exhaustion-related genes differentially expressed between PD-1^KD BCMA CAR+ and BCMA CAR+T cells after four repetitive stimulations (d13). The results are from two representative healthy donors with two duplicates. BCMA, B cell maturation antigen; GSEA, gene set enrichment analysis; HD, heathy donor; NES, normalized enrichment score; FDR, false discovery rate.