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生成抗同种异体排斥通用型嵌合抗原受体自然杀伤T细胞并评估其疗效、作用机制、安全性和免疫原性的方案。

Protocol to generate allorejection-resistant universal CAR-NKT cells and evaluate their efficacy, mechanism of action, safety, and immunogenicity.

作者信息

Fang Ying, Chen Yuning, Zhu Yichen, Tian Yanxin, Li Yan-Ruide, Yang Lili

机构信息

Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.

Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095, USA.

出版信息

STAR Protoc. 2025 May 2;6(2):103810. doi: 10.1016/j.xpro.2025.103810.

DOI:10.1016/j.xpro.2025.103810
PMID:40317722
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12090235/
Abstract

Universal chimeric antigen receptor-engineered invariant natural killer T (CAR-NKT) cells overcome the limitations of allogeneic chimeric antigen receptor (CAR)-T cell therapies, such as graft-versus-host disease and allorejection. Here, we present a protocol for generating CAR-NKT cells from hematopoietic stem and progenitor cells (HSPCs), followed by feeder-free ex vivo differentiation. We also outline assays to assess antitumor efficacy, safety, and allorejection resistance. For complete details on the use and execution of this protocol, please refer to Li et al..

摘要

通用嵌合抗原受体工程化不变自然杀伤T细胞(CAR-NKT细胞)克服了同种异体嵌合抗原受体(CAR)-T细胞疗法的局限性,如移植物抗宿主病和同种异体排斥反应。在此,我们展示了一种从造血干细胞和祖细胞(HSPCs)生成CAR-NKT细胞的方案,随后进行无饲养层的体外分化。我们还概述了评估抗肿瘤疗效、安全性和同种异体排斥抗性的检测方法。有关本方案使用和执行的完整详细信息,请参考Li等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/134ee7e04cfa/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d75a61cc1987/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d4aea2952dc4/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/7024e56010e2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d5b8d7230141/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/2d0eeeb1fdf0/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/134ee7e04cfa/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d75a61cc1987/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d4aea2952dc4/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/7024e56010e2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/d5b8d7230141/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/2d0eeeb1fdf0/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4343/12090235/134ee7e04cfa/gr5.jpg

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本文引用的文献

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Nat Commun. 2025 Feb 1;16(1):1248. doi: 10.1038/s41467-025-56270-6.
2
Generating allogeneic CAR-NKT cells for off-the-shelf cancer immunotherapy with genetically engineered HSP cells and feeder-free differentiation culture.利用基因工程改造的热休克蛋白(HSP)细胞和无饲养层分化培养技术生成用于现货型癌症免疫治疗的同种异体嵌合抗原受体自然杀伤T细胞(CAR-NKT细胞)
Nat Protoc. 2025 May;20(5):1352-1388. doi: 10.1038/s41596-024-01077-w. Epub 2025 Jan 17.
3
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使用临床指导的培养方法从造血干细胞和祖细胞生成同种异体CAR-NKT细胞。
Nat Biotechnol. 2025 Mar;43(3):329-344. doi: 10.1038/s41587-024-02226-y. Epub 2024 May 14.
4
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Mol Ther. 2024 Jun 5;32(6):1849-1874. doi: 10.1016/j.ymthe.2024.04.005. Epub 2024 Apr 6.
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Cell Rep Med. 2021 Nov 16;2(11):100449. doi: 10.1016/j.xcrm.2021.100449.
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