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| Item Type: | Article | ||||
|---|---|---|---|---|---|
| Title: | TCR gene therapy for cancer | ||||
| Creators Name: | Rhein, S. and Çakmak-Görür, N. | ||||
| Abstract: | 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. | ||||
| Keywords: | TCR, T Cell Stimulation, Antigen-Specific T Cells, T Cell Sort, RACE PCR, Viral Transduction, Animals, Mice | ||||
| Source: | Methods in Molecular Biology | ||||
| Series Name: | Methods in Molecular Biology | ||||
| Title of Book: | Gene therapy of cancer : methods and protocols | ||||
| ISSN: | 1064-3745 | ||||
| ISBN: | 9781071624401 | ||||
| Publisher: | Springer / Humana Press | ||||
| Volume: | 2521 | ||||
| Number: | 3 | ||||
| Page Range: | 95-128 | ||||
| Number of Pages: | 445 | ||||
| Date: | 23 June 2022 | ||||
| Official Publication: | https://doi.org/10.1007/978-1-0716-2441-8_6 | ||||
| PubMed: | View item in PubMed | ||||
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