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

. 2021 Nov 8;12:6446. doi: 10.1038/s41467-021-26844-1

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. 6 — Eight- to 12-week-old male NSG mice were conditioned with busulfan on days −2 and −1 before IV injection of 5.10⁶ HLA-A2+ PBMCs followed by retroorbital injection of 5 × 10⁶ of Tregs (b–d): PBMC group (n = 16), CD28 CAR-Tregs (n = 14), 4-1BB CAR-Tregs (n = 10), 4-1BB + rapamycin/vitamin c (n = 13), and MOCK-Tregs (n = 8) from 4, 4, 1, 3, and 2 independent experiments, respectively. a Schematic design of the experiment. Mice were weighed and scored for GVHD three times a week and bled weekly for flow-cytometry analysis. b Survival curves (left panel) and GVHD scores (right panel). Log-rank Mantel-Cox tests comparing the indicated groups. Both CD28 and 4-1BB CAR-Tregs + rapamycin/vitamin c groups demonstrate a significantly better survival than the PBMC group (****p < 0.0001). c Plasma human IFN-γ levels were measured by cytometry bead array at days 10, 17, and 24. IFN-γ levels are significantly lower in CAR-Treg groups than in the PBMC group at day 17 (CD28, ***p = 0,0004; 4-1BB, *p = 0,0147, 4-1BB + rapamycin/vitamin C, *p = 0,0147) and at day 24 (CD28, ***p = 0,0002 and 4-1BB + rapamycin/vitamin C, **p = 0,0073). d Representative contour plots of circulating murine and human cells and frequencies of transferred PBMCs (lower panels). Compared to PBMC group, chimerism is statistically different at days 10 (4-1BB, ****p < 0,0001), 17 (MOCK, *p = 0,0274; CD28, **p = 0,0060; 4-1BB + rapamycin/vitamin C, ***p = 0,0005) and 24 (CD28, ****p < 0,0001; 4-1BB + rapamycin/vitamin C, **p = 0,0030). e Histologic analysis of liver and lung lesions at sacrifice from three independent experiments and three different donors, except for MOCK-Tregs (two experiments) and 4-1BB CAR-Tregs (one experiment). PBMCs only (n = 9), CD28 CAR-Tregs (n = 11), 4-1BB CAR-Tregs (n = 10), 4-1BB CAR-Tregs + rapamycin/ vitamin C (n = 10), and MOCK-Tregs (n = 4). Statistical significance is observed in the following comparisons: PBMC vs. 4-1BB + rapamycin/vitamin C, p < 0.0001 (liver) and p = 0.0346 (lung); PBMC vs. CD28, p = 0,0076 (liver) and p = 0,0043 (lung); PBMC vs. 4-1BB, p = 0,04 (liver) and p = 0.0031 (lung); and 4-1BB cultured with or without rapamycin/vitamin C p = 0,0002 (liver). Data are depicted as mean +/− SEM (panels b to e) and compared using two-tailed Mann–Whitney test (c to e) (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). Abbreviations: B, busulfan; L, bioluminescence.