CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
癌症的 TCR 基因治疗
英文原题：TCR Gene Therapy for Cancer.
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TCR Gene Therapy for Cancer.
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中文摘要
该方案描述了用于过继性 T 细胞治疗的抗原特异性 T 细胞生成和 TCR 鉴定的程序。我们描述了生成抗原特异性 T 细胞的两种途径：第一，用负载抗原肽的自体树突状细胞刺激 T 细胞；第二，在合适的 mouse 模型中通过基因枪应用肽或 DNA 进行体内 T 细胞刺激，随后体外富集肽反应性 T 细胞。经肽刺激的 T 细胞通过荧光激活细胞分选术按 CD107α 或 IFNγ 表达进行分选，随后分离的 RNA 用于针对 TCR 的 5' cDNA 末端快速扩增（RACE）-PCR，以进行 TCR 链鉴定。经过逆转录病毒克隆后，将其重新表达于人类 T 细胞上，以测试其在过继性 T 细胞治疗中的适用性。
展开英文摘要原文
The protocol describes the procedure of antigen-specific T cell generation and TCR identification for the use in adoptive T cell therapy.
We describe two paths of generating antigen-specific T cells, first, T cell stimulation with autologous dendritic cells pulsed with antigen peptide, second, in vivo T cell stimulation with peptide or DNA by gene gun application in a suitable mouse model followed by in vitro enrichment of peptide-reactive T cells.
Peptide-stimulated T cells are sorted by fluorescence-activated cell sorting for CD107α or IFNγ expression and subsequently isolated RNA is used in a 5' rapid amplification of cDNA ends (RACE )-PCR specific for TCR for TCR chain identification. After retroviral cloning, it is re-expressed on human T cells to test its applicability in adoptive T cell therapy.
论文信息
- 作者
- Rhein S、Çakmak-Görür N
- 单位
- Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany. simone.rhein@mdc-berlin.de.Germany
- 期刊
- Methods in molecular biology (Clifton, N.J.)2022
- 原文标识
- PubMed 35732995 · DOI 10.1007/978-1-0716-2441-8_6