TCR gene therapy for cancer

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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
Related to:	URL URL Type https://edoc.mdc-berlin.de/21792/ Book

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