Department of Biological Sciences, Clemson University, Clemson, SC, United States of America.
PLoS One. 2018 Jul 25;13(7):e0201100. doi: 10.1371/journal.pone.0201100. eCollection 2018.
Many of the cytokine-based cancer immunotherapies are hindered by the devastating side effects of systemic delivery of the cytokines. To address this problem, we previously described a novel approach to locally achieve high doses of interleukin-12 (IL-12) in tumors and demonstrated that bi-functional fusion protein mIL-12/FasTI expressed by stable clones of TC-1 cells efficiently suppressed tumor proliferation by activating natural killer (NK) cells and other cytolytic killer cells and sending apoptotic signals into tumor cells. In the present study, we employed a lentiviral vector-based gene delivery system to deliver this fusion construct directly into tumor cells. We show that lentiviral vector efficiently delivers the fusion constructs into Hela cells in vitro as assayed by RT-PCR and immunohistochemistry (IHC). We also confirm that fusion protein mIL-12/FasTI delivered by the viral vector significantly enhanced killer cell activation, increased caspase-3 activity and decreased tumor growth in vitro. This study offers a further step for fusion protein cancer therapy for cancer patients.
许多基于细胞因子的癌症免疫疗法受到细胞因子全身给药的破坏性副作用的阻碍。为了解决这个问题，我们之前描述了一种在肿瘤中局部获得高剂量白细胞介素-12 (IL-12) 的新方法，并证明由 TC-1 细胞稳定克隆表达的双功能融合蛋白 mIL-12/FasTI 通过激活自然杀伤 (NK) 细胞和其他细胞毒性杀伤细胞，并向肿瘤细胞发送凋亡信号，有效地抑制了肿瘤的增殖。在本研究中，我们使用基于慢病毒载体的基因传递系统将这种融合构建体直接递送至肿瘤细胞。我们通过 RT-PCR 和免疫组织化学 (IHC) 证实，慢病毒载体可有效地将融合构建体递送至体外的 Hela 细胞中。我们还证实，病毒载体递送的融合蛋白 mIL-12/FasTI 可显著增强杀伤细胞的激活、增加 caspase-3 活性并减少体外肿瘤生长。这项研究为癌症患者的融合蛋白癌症治疗提供了进一步的步骤。
