背景：嵌合抗原受体 (CAR) T 细胞疗法可用于治疗造血起源的癌症，但实体瘤中的嵌合抗原受体 (CAR) T 细胞疗法会因 CAR 识别的抗原丢失而影响疗效。然而，树突状细胞 (DC)/肿瘤融合疫苗呈现一系列已知或未知的肿瘤抗原，以刺激 T 细胞扩增并增强 T 细胞反应。通过 DC/肿瘤融合疫苗刺激来开发增强基于纳米抗体的 CAR-T (Nb-CAR-T) 细胞抗肿瘤活性的新策略将为更有效的 CAR-T 细胞疗法提供指导。方法：考虑到纳米抗体（Nb）的治疗潜力，我们首先筛选EGFRvIII Nb，然后构建并验证EGFRvIII Nb-CAR-T细胞的体外和体内功能。我们进一步联合DC/肿瘤融合疫苗来增强EGFRvIII Nb-CAR-T细胞的抗肿瘤效果，并在体外和荷瘤异种移植小鼠模型中评估了Nb-CAR-T细胞的功能。结果：我们首次成功选择 EGFRvIII Nb 来生成新型 EGFRvIII Nb-CAR-T 细胞。重要的是，我们的结果表明，DC/肿瘤融合疫苗刺激 Nb-CAR-T 细胞反应不仅可以改善体外 T 细胞增殖、T 细胞活化、细胞因子分泌和肿瘤特异性细胞毒性，而且还可以显着减轻肿瘤负荷，延长生存期并改善 Nb-CAR-T 细胞浸润。结论：我们创新性地证明，DC/肿瘤融合疫苗可显着增强 Nb-CAR-T 细胞对抗实体瘤的功效。这一新策略为促进CAR-T细胞疗法的临床治疗提供了一个有前景的治疗平台。©作者。

Background: Chimeric antigen receptor (CAR) T-cell therapy is practical in treating cancers of hematopoietic origin, but of that in solid tumors compromises efficacy for the loss of the antigen recognized by the CAR. However, dendritic cell (DC)/tumor fusion vaccines present a spectrum of known or unknown tumor antigens to stimulate T cell expansion and enhanced T cell response. Developing a new strategy of enhanced nanobody-based CAR-T (Nb-CAR-T) cells antitumor activity by DC/tumor fusion vaccines stimulation would provide guidance for more effective CAR-T cell therapies. Methods: Considering the therapeutic potential of nanobody (Nb), we first screened EGFRvIII Nb, then constructed and verified the function of EGFRvIII Nb-CAR-T cells in vitro and in vivo. We further combined DC/tumor fusion vaccines to boost EGFRvIII Nb-CAR-T cells antitumor effect, which was evaluated in vitro Nb-CAR-T cell function and in the tumor-bearing xenograft mouse models. Results: We had for the first time successfully selected EGFRvIII Nb for the generation of the novel EGFRvIII Nb-CAR-T cells. Importantly, our results suggested that DC/tumor fusion vaccines stimulate Nb-CAR-T cells response not only in improving T cell proliferation, T cell activation, cytokine secretion and tumor-specific cytotoxicity in vitro, but also significantly reducing tumor burden, prolonging survival and improving Nb-CAR-T cells infiltration. Conclusions: We have innovatively shown that DC/tumor fusion vaccines significantly enhance the efficacy of Nb-CAR-T cells against solid tumors. This new strategy has provided a promising therapeutic platform for promoting the clinical treatment of CAR-T cells therapy.© The author(s).