Department of Neurosurgery, Cedars-Sinai Medical Center, 127 S. San Vicente Boulevard Suite A6600, Los Angeles, CA, 90048, USA.
Department of Radiation Oncology, Cedars-Sinai Medical Center, 8700 Beverly Boulevard, Los Angeles, CA, 90048, USA.
Sci Rep. 2023 Jan 5;13(1):196. doi: 10.1038/s41598-022-27184-w.
Naïve T cells become effector T cells following stimulation by antigen-loaded dendritic cells (DCs) and sequential cytokine activation. We aimed to develop procedures to efficiently activate T cells with tumor-associated antigens (TAAs) to glioblastoma (GBM) stem cells. To remove antigen presentation outside of the immunosuppressive tumor milieu, three different glioma stem cell (GSC) specific antigen sources to load DCs were compared in their ability to stimulate lymphocytes. An activated T cell (ATC) protocol including cytokine activation and expansion in culture to target GSCs was generated and optimized for a planned phase I clinical trial. We compared three different antigen-loading methods on DCs to effectively activate T cells, which were GBM patient-derived GSC-lysate, acid-eluate of GSCs and synthetic peptides derived from proteins expressed in GSCs. DCs derived from HLA-A2 positive blood sample were loaded with TAAs. Autologous T cells were activated by co-culturing with loaded DCs. Efficiency and cytotoxicity of ATCs were evaluated by targeting TAA-pulsed DCs or T2 cells, GSCs, or autologous PHA-blasts. Characteristics of ATCs were evaluated by Flow Cytometry and ELISpot assay, which showed increased number of ATCs secreting IFN-γ targeting GSCs as compared with non-activated T cells and unloaded target cells. Neither GSC-lysate nor acid-eluate loading showed enhancement in response of ATCs but the synthetic peptide pool showed significantly increased IFN-γ secretion and increased cytotoxicity towards target cells. These results demonstrate that ATCs activated using a TAA synthetic peptide pool efficiently enhance cytotoxicity specifically to target cells including GSC.
幼稚 T 细胞在抗原负载树突状细胞 (DC) 的刺激下和顺序细胞因子激活后成为效应 T 细胞。我们旨在开发有效激活针对脑胶质瘤（GBM）干细胞的肿瘤相关抗原 (TAA) 的 T 细胞的程序。为了消除免疫抑制肿瘤微环境之外的抗原呈递，我们比较了三种不同的脑胶质瘤干细胞 (GSC) 特异性抗原来源，以比较其刺激淋巴细胞的能力。生成了一种包括细胞因子激活和培养中扩增以靶向 GSCs 的激活 T 细胞 (ATC) 方案，并针对计划中的 I 期临床试验进行了优化。我们比较了三种不同的抗原加载方法对 DC 有效激活 T 细胞的能力，这三种方法是 GBM 患者来源的 GSC 裂解物、GSCs 的酸洗脱物和源自 GSCs 中表达的蛋白质的合成肽。从 HLA-A2 阳性血液样本中提取的 DC 加载 TAA。通过与负载的 DC 共培养来激活自体 T 细胞。通过靶向 TAA 脉冲的 DC 或 T2 细胞、GSCs 或自体 PHA 母细胞来评估 ATC 的效率和细胞毒性。通过流式细胞术和 ELISpot 分析评估 ATC 的特征，结果表明，与非激活的 T 细胞和未负载的靶细胞相比，针对 GSCs 的 ATC 分泌 IFN-γ 的数量增加。GSC 裂解物或酸洗脱物加载均未增强 ATC 的反应，但合成肽库显示出明显增加的 IFN-γ 分泌和对靶细胞的增加的细胞毒性。这些结果表明，使用 TAA 合成肽库激活的 ATC 可有效增强针对包括 GSC 在内的靶细胞的细胞毒性。
