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. Author manuscript; available in PMC: 2025 Jul 29.
Published in final edited form as: Nat Cancer. 2025 May 16;6(6):1073–1087. doi: 10.1038/s43018-025-00968-5

Fig. 5 |. Adoptive transfer of anti-VHH CAR T cells into mice with VHH-expressing tumors delays tumor growth, enhances infiltration of tumor-reactive T cells and promotes endogenous antitumor immunity.

Fig. 5 |

a, MC38-VHH tumors were treated with murine anti-VHH CAR T cells and monitored for tumor growth. T cells were isolated from tumors, spleens and lymph nodes on day 18 for downstream analysis. s.c., subcutaneous; LN, lymph node; TCR, T cell receptor. b, Tumor growth curves following ACT. Statistical analysis was performed using a two-way ANOVA (mean ± s.e.m., n = 4 mice). **P = 0.0043 and ****P < 0.0001. c, Count and frequency of indicated T cell populations isolated from the tumor. Statistical analysis was performed using an unpaired one-sided Student’s t-test (mean displayed, n = 4 tumors for control CAR T cells and n = 5 tumors for anti-VHH CAR T cells). Left, **P = 0.0013; middle, **P = 0.0016; right, *P = 0.0492. d, Representative flow plot (n = 5 for anti-VHH CAR T cells and n = 4 for control CAR T cells) of endogenous (CAR−) T cell expression of the Reps1 T cell receptor in tdLNs and the frequency of this population in the ipsilateral and contralateral draining lymph nodes. Statistical analysis was performed using an unpaired two-sided Student’s t-test (mean displayed, n = 3 independent lymph nodes for contralateral control CAR T cell, n = 4 independent lymph nodes for ipsilateral control CAR T cell and contralateral anti-VHH CAR T cell and n = 5 independent lymph nodes for ipsilateral anti-VHH CAR T cell. *P = 0.0374. NS, nonsignificant. Each lymph node was obtained from an individual mouse. e, E0771-VHH tumors were treated with murine anti-VHH CAR T cells. f, Individual traces of tumor growth curves of E0771-VHH tumor-bearing mice treated with control CAR T cells (black) or anti-VHH CAR T cells (blue) (n = 4 mice). g, Next, 45 days after initial treatment, cured mice (n = 3) were rechallenged with WT E0771 tumor cells. Statistical analysis was performed using a two-way ANOVA (mean ± s.e.m., n = 5 mice). ****P < 0.0001. h, Survival curves of tumor-bearing mice following treatment. Statistical analysis was performed using a log-rank (Mantel–Cox) test (n = 4 mice). **P = 0.0019.