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TCR gene therapy for cancer

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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