Generation of a Jurkat-based fluorescent reporter cell line to evaluate lipid antigen interaction with the human iNKT cell receptor
PublisherNature Publishing Group
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AbstractInvariant natural killer T (iNKT) cells are a specialized subset of T cells contributing to both, the innate and adaptive immune responses. In contrast to conventional T lymphocytes they recognize lipid antigens. The aim of the project is to establish a novel model system, to study iNKT-TCR – ligand interaction. An iNKT reporter cell line (JE6-1 REP-iNKT ) was engineered by introducing the human iNKT-TCR into a human leukemic T cell line carrying an NF-κB-driven fluorescent transcriptional reporter construct. Antigen presenting BW STIM cells expressing human CD1d and CD80 were generated. Reporter induction in JE6-1 REP-iNKT cells was assessed by flow cytometry. CRISPR/Cas9 was used for β2M knock out in JE6-1 REP-iNKT cells to abrogate CD1d expression and thus excluding antigen self-presentation. Reporter cells were shown to specifically react with iNKT antigens presented via CD1d. Their sensitivity towards α-GalCer was comparable to a murine iNKT hybridoma cell line. In conclusion, we created a novel iNKT reporter platform which, compared to traditional iNKT cell assays, is characterized by a shorter turnaround time and lower costs. It thus facilitates the identification of antigenic structures that drive the activation of iNKT cells in health and disease. © 2019, The Author(s).
SponsorsTis work was partially funded by the Austrian Science Fund (FWF): DK W 1248-B13 for PH and SFB F4603 for KHS.
Rights/TermsAttribution 3.0 United States
Identifier to cite or link to this itemhttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85065767249&origin=inward; http://hdl.handle.net/10713/9327
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