Chimeric antigen receptor T (CAR-T) cell therapy holds great promise as an innovative immunotherapeutic approach for cancer treatment. To optimize the production and application of CAR-T cells, we evaluated the in vivo stability and efficacy capacities of CAR-T cells developed under different conditions. In this study, CAR-T cells were activated using Phytohemagglutinin (PHA) or anti-CD3&anti-CD28 and were compared in an in vivo CD19+B-cell cancer model in mouse groups. Our results demonstrated that CAR-T cells activated with PHA exhibited higher stability and anti-cancer efficacy compared to those activated with anti-CD3&anti-CD28. Specifically, CAR19BB-T cells activated with PHA exhibited continuous proliferation and long-term persistence without compromising their anti-cancer efficacy. Kaplan–Meier survival analysis revealed prolonged overall survival in the CAR-T cell-treated groups compared to the only tumor group. Furthermore, specific LTR-targeted RT-PCR analysis confirmed the presence of CAR-T cells in the treated groups, with significantly higher levels observed in the CAR19BB-T (PHA) group compared to other groups. Histopathological analysis of spleen, kidney, and liver tissue sections indicated reduced inflammation and improved tissue integrity in the CAR-T cell-treated groups. Our findings highlight the potential benefits of using PHA as a co-stimulatory method for CAR-T cell production, offering a promising strategy to enhance their stability and persistence. These results provide valuable insights for the development of more effective and enduring immunotherapeutic approaches for cancer treatment. CAR-T cells activated with PHA may offer a compelling therapeutic option for advancing cancer immunotherapy in clinical applications.

嵌合抗原受体 T (CAR-T) 细胞疗法作为一种创新的癌症免疫治疗方法具有广阔的前景。为了优化 CAR-T 细胞的生产和应用，我们评估了在不同条件下开发的 CAR-T 细胞的体内稳定性和功效能力。在本研究中，使用植物血凝素 (PHA) 或抗 CD3 和抗 CD28 激活 CAR-T 细胞，并在小鼠组的体内 CD19+B 细胞癌症模型中进行比较。我们的结果表明，与抗CD3和抗CD28激活的CAR-T细胞相比，用PHA激活的CAR-T细胞表现出更高的稳定性和抗癌功效。具体来说，用PHA激活的CAR19BB-T细胞表现出持续增殖和长期持久性，且不影响其抗癌功效。Kaplan-Meier 生存分析显示，与单一肿瘤组相比，CAR-T 细胞治疗组的总体生存期延长。此外，特定的 LTR 靶向 RT-PCR 分析证实了治疗组中存在 CAR-T 细胞，与其他组相比，CAR19BB-T (PHA) 组中观察到的水平显着更高。脾脏、肾脏和肝脏组织切片的组织病理学分析表明，CAR-T 细胞治疗组的炎症减轻并改善了组织完整性。我们的研究结果强调了使用 PHA 作为 CAR-T 细胞生产的共刺激方法的潜在好处，为增强其稳定性和持久性提供了一种有前景的策略。这些结果为开发更有效和持久的癌症治疗免疫治疗方法提供了宝贵的见解。用 PHA 激活的 CAR-T 细胞可能为临床应用中推进癌症免疫疗法提供令人信服的治疗选择。