单细胞追踪揭示黑色素瘤 TIL 治疗过程中肿瘤反应性 T 细胞的可塑性
Single-cell tracking reveals tumor-reactive T cell plasticity during melanoma TIL therapy.
TIL(肿瘤浸润淋巴细胞)过继细胞治疗可在转移性黑色素瘤中诱导持久缓解,然而在体外扩增过程中及回输后,调控肿瘤反应性T细胞命运的克隆和转录动态仍知之甚少。
英文原题:Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer.
胰腺癌是一种侵袭性恶性肿瘤,预后极差,治疗选择有限。
胰腺癌是一种侵袭性恶性肿瘤,预后极差且治疗选择有限。过继性细胞疗法是一种有前景的实验性方法,该疗法涉及分离并激活患者自身的免疫细胞,如TIL(肿瘤浸润淋巴细胞)(TILs),然后再回输。然而,在临床前胰腺癌模型中,过继性细胞转移的技术尚未充分建立。在此,我们描述了一种使用来自同基因胰腺癌小鼠模型的TILs进行过继性细胞疗法的详细方案。该流程包括将活的或经照射的小鼠胰腺癌细胞植入荧光标记的报告小鼠中以启动免疫细胞浸润,然后通过流式细胞术分选从原发肿瘤中分离淋巴细胞,和/或在体外激活并扩增肿瘤反应性T细胞,并将这些激活的T细胞腹腔过继转移至荷瘤小鼠,随后给予白细胞介素-2。生物发光肿瘤成像可纵向监测原位肿瘤生长及对治疗的反应,特别是评估肿瘤特异性细胞毒性效应。该方法重现了为胰腺癌患者开发过继性细胞转移疗法所涉及的流程。结果表明,与无关淋巴细胞对照相比,过继转移的肿瘤反应性T细胞具有增强的抗肿瘤疗效。这种多功能方法使得能够在体内研究胰腺癌的过继性免疫治疗,并优化细胞处理参数和联合治疗方案。
Pancreatic cancer is an aggressive malignancy with a dismal prognosis and limited therapeutic options. Adoptive cell therapy, which involves isolating and activating a patient's own immune cells, such as tumor-infiltrating lymphocytes (TILs), before re-infusing them, represents a promising experimental approach. However, techniques for adoptive cell transfer in preclinical pancreatic cancer models are not well established. Here, we describe a detailed protocol for adoptive cell therapy using TILs from a syngeneic pancreatic cancer mouse model. The procedure involves implanting live or irradiated mouse pancreatic cancer cells in fluorescence-labeled reporter mice to initiate immune cell influx, then isolating lymphocytes from primary tumors via flow cytometry sorting and/or activating and expanding tumor-reactive T cells ex vivo, and adoptively transferring these activated T cells intraperitoneally into tumor-bearing mice, followed by interleukin-2 administration. Bioluminescent tumor imaging allows for longitudinal monitoring of orthotopic tumor growth and response to therapy, especially evaluating the tumor-specific cytotoxic effects. This approach recapitulates the logistics involved in developing adoptive cell transfer therapies for pancreatic cancer patients. The results demonstrate enhanced antitumor efficacy of adoptively transferred tumor-reactive T cells compared to irrelevant lymphocyte controls. This versatile methodology enables the in vivo study of adoptive immunotherapy in pancreatic cancer as well as the optimization of cell processing parameters and combination treatment regimens.
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