Melanoma Immunotherapy in Mice Using Genetically Engineered Pluripotent Stem Cells.
Melanoma Immunotherapy in Mice Using Genetically Engineered Pluripotent Stem Cells.
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DOI：
10.3727/096368916x690467
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发表时间：
2016
影响因子：
3.3
通讯作者：
Song J
中科院分区：
文献类型：
--
作者：
Haque M;Song J;Fino K;Sandhu P;Wang Y;Ni B;Fang D;Song J
抗原（Ag）特异性CD 8+细胞毒性T淋巴细胞（CTL）的连续细胞转移（ACT）是用于多种疾病的非常有前途的治疗。幼稚或中枢记忆T细胞衍生的效应CTL是基于ACT的免疫疗法的最佳群体，因为这些细胞具有高增殖潜力，比终末分化细胞更不易于凋亡，并且具有更高的响应稳态细胞因子的能力。然而，由于难以从患者获得足够的细胞，这种具有T细胞持久性的ACT通常是不可行的。在这里，我们提出，在体外分化的HSC的工程PSC可以在体内发展成肿瘤Ag特异性幼稚CTL，有效地抑制黑色素瘤的生长。小鼠诱导的PSC（iPSC）用编码鸡卵清蛋白（OVA）特异性T细胞受体（TCR）和存活相关蛋白（即，BCL-xL和生存素）。在鼠重组细胞因子（rFlt 3L和rIL-7）的存在下，将基因转导的iPSC在表达δ样配体1的0 P9（0 P9-DL 1）鼠基质细胞上培养一周。然后将这些iPSC衍生的细胞静脉内过继转移到受体小鼠中，随后腹膜内注射激动剂α-Notch 2抗体和细胞因子（rFlt 3L和rIL-7）。两周后，在小鼠外周淋巴系统中观察到幼稚OVA特异性CD 8 + T细胞，其对OVA特异性刺激有反应。此外，小鼠对B16-OVA黑色素瘤诱导的挑战具有抵抗力。这些结果表明，遗传修饰的干细胞可用于基于ACT的免疫治疗或作为潜在的疫苗。
Adoptive cell transfer (ACT) of antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) is a highly promising treatment for a variety of diseases. Naive or central memory T-cell-derived effector CTLs are optimal populations for ACT-based immunotherapy because these cells have a high proliferative potential, are less prone to apoptosis than terminally differentiated cells, and have the higher ability to respond to homeostatic cytokines. However, such ACT with T-cell persistence is often not feasible due to difficulties in obtaining sufficient cells from patients. Here we present that in vitro differentiated HSCs of engineered PSCs can develop in vivo into tumor Ag-specific naive CTLs, which efficiently suppress melanoma growth. Mouse-induced PSCs (iPSCs) were retrovirally transduced with a construct encoding chicken ovalbumin (OVA)-specific T-cell receptors (TCRs) and survival-related proteins (i.e., BCL-xL and survivin). The gene-transduced iPSCs were cultured on the delta-like ligand 1-expressing OP9 (OP9-DL1) murine stromal cells in the presence of murine recombinant cytokines (rFlt3L and rIL-7) for a week. These iPSC-derived cells were then intravenously adoptively transferred into recipient mice, followed by intraperitoneal injection with an agonist α-Notch 2 antibody and cytokines (rFlt3L and rIL-7). Two weeks later, naive OVA-specific CD8+ T cells were observed in the mouse peripheral lymphatic system, which were responsive to OVA-specific stimulation. Moreover, the mice were resistant to the challenge of B16-OVA melanoma induction. These results indicate that genetically modified stem cells may be used for ACT-based immunotherapy or serve as potential vaccines.