Research Papers:

A Jurkat 76 based triple parameter reporter system to evaluate TCR functions and adoptive T cell strategies

Sandra Rosskopf, Judith Leitner, Wolfgang Paster, Laura T. Morton, Renate S. Hagedoorn, Peter Steinberger and Mirjam H.M. Heemskerk _

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Oncotarget. 2018; 9:17608-17619. https://doi.org/10.18632/oncotarget.24807

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Sandra Rosskopf1, Judith Leitner1, Wolfgang Paster1, Laura T. Morton2, Renate S. Hagedoorn2, Peter Steinberger1 and Mirjam H.M. Heemskerk2

1Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

2Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands

Correspondence to:

Mirjam H.M. Heemskerk, email: [email protected]

Peter Steinberger, email: [email protected]

Keywords: adoptive T cell therapy; TCR gene transfer; immunotherapy; reporter T cells; chimeric receptors

Received: November 22, 2017     Accepted: February 26, 2018     Published: April 03, 2018


Adoptive T cell therapy using TCR transgenic autologous T cells has shown great potential for the treatment of tumor patients. Thorough characterization of genetically reprogrammed T cells is necessary to optimize treatment success. Here, we describe the generation of triple parameter reporter T cells based on the Jurkat 76 T cell line for the evaluation of TCR and chimeric antigen receptor functions as well as adoptive T cell strategies. This Jurkat subline is devoid of endogenous TCR alpha and TCR beta chains, thereby circumventing the problem of TCR miss-pairing and unexpected specificities. The resultant reporter cells allow simultaneous determination of the activity of the transcription factors NF-κB, NFAT and AP-1 that play key roles in T cell activation. Human TCRs directed against tumor and virus antigens were introduced and reporter responses were determined using tumor cell lines endogenously expressing the antigens of interest or via addition of antigenic peptides. Finally, we demonstrate that coexpression of adhesion molecules like CD2 and CD226 as well as CD28 chimeric receptors represents an effective strategy to augment the response of TCR-transgenic reporters to cells presenting cognate antigens.

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