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. 2024 Nov 25;14(1):2432345. doi: 10.1080/22221751.2024.2432345

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© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 7. — Evaluation of in vitro CAR-T cell persistence. (A) Schematic representation of rechallenge experiment time course. CAR-T cells were first exposed to target cells (ratio 5:1) and allowed to engage and eliminate the targets over a period of 24 h. Post initial challenge, CAR-T cells were re-isolated and re-challenged with a fresh batch of target cells three times (2nd, 3rd and 4th challenge). (B) Graph shows grouped analysis of CAR-T cell cytotoxicity during 4th challenge. Values are from 5 independent measurements. (C) Proliferation of CAR-T cells during the rechallenge experiment evaluated by total count of CD3⁺ T cells by flow cytometry. Values are from 4 independent measurements. (D-F) Exhaustion profile of the different CAR-T cells after 4 challenges evaluated by flow cytometry staining of PD-1, CTLA-4 and LAG-3 exhaustion markers. Values are from 4 independent measurements. Significance was assigned as follows: ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.