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An immunoproteomic approach to characterize the CAR interactome and signalosome.

Sci Signal. 2019-02; 
RamelloMaria C,Benza?dIsmahène,KuenziBrent M,Lienlaf-MorenoMaritza,KandellWendy M,SantiagoDaniel N,Pabón-Salda?aMibel,DarvilleLancia,FangBin,RixUwe,YoderSean,BerglundAnders,KoomenJohn M,HauraEric B,Abate-DagaDa
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Recombinant Antibody Services CAR expression on transduced lymphocytes was detected by using biotinylated Protein L (GenScript) and a secondary incubation with Streptavidin PE (Biolegend). Get A Quote

摘要

Adoptive transfer of T cells that express a chimeric antigen receptor (CAR) is an approved immunotherapy that may be curative for some hematological cancers. To better understand the therapeutic mechanism of action, we systematically analyzed CAR signaling in human primary T cells by mass spectrometry. When we compared the interactomes and the signaling pathways activated by distinct CAR-T cells that shared the same antigen-binding domain but differed in their intracellular domains and their in vivo antitumor efficacy, we found that only second-generation CARs induced the expression of a constitutively phosphorylated form of CD3ζ that resembled the endogenous species. This phenomenon was independent of the... More

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