Oncotarget

Research Papers: Immunology:

A cellular platform for the evaluation of immune checkpoint molecules

Sabrina Jutz, Annika Hennig, Wolfgang Paster, Ömer Asrak, Dejana Dijanovic, Florian Kellner, Winfried F. Pickl, Johannes B. Huppa, Judith Leitner, Peter Steinberger _

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Oncotarget. 2017; 8:64892-64906. https://doi.org/10.18632/oncotarget.17615

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Abstract

Sabrina Jutz1,*, Annika Hennig1,*, Wolfgang Paster1, Ömer Asrak1, Dejana Dijanovic1, Florian Kellner2, Winfried F. Pickl3, Johannes B. Huppa2, Judith Leitner1 and Peter Steinberger1

1 Division of Immune Receptors and T cell Activation, Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

2 Department of Molecular Immunology, Immune Recognition Unit, Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

3 Division of Cellular Immunology and Immunohematology, Institute of Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria

* These authors have contributed equally to this manuscript and share first authorship

Correspondence to:

Peter Steinberger, email:

Keywords: T cell activation, NF-κB, reporter cell lines, Jurkat, immune checkpoints

Received: November 14, 2016 Accepted: April 22, 2017 Published: May 04, 2017

Abstract

Blockade of the T cell coinhibitory molecules CTLA-4 and PD-1 has clinical utility to strengthen T cell responses. In addition to these immune checkpoints an ever-growing number of molecules has been implicated in generating coinhibitory signals in T cells. However, investigating coinhibitory molecules in primary human cells is complicated by the restricted expression and promiscuity of both coinhibitory receptors and their ligands. Here we have evaluated the potential of fluorescence-based transcriptional reporters based on the human Jurkat T cell line in conjunction with engineered T cell stimulator cell lines for investigating coinhibitory pathways. CTLA-4, PD-1, TIGIT, BTLA and 2B4 expressing reporter cells were generated and activated with T cell stimulator cells expressing cognate ligands of these molecules.

All accessory molecules tested were functional in our reporter system. Engagement of CTLA-4, PD-1, BTLA and TIGIT by their ligands significantly inhibited T cell activation, whereas binding of 2B4 by CD48 resulted in enhanced responses. Mutational analysis revealed intracellular motifs that are responsible for BTLA mediated T cell inhibition and demonstrates potent reporter inhibition by CTLA-4 independent of cytoplasmic signaling motifs. Moreover, considerably higher IC50 values were measured for the CTLA-4 blocker Ipilimumab compared to the PD-1 antibody Nivolumab.

Our findings show that coinhibitory pathways can be evaluated in Jurkat-based transcriptional reporters and yield novel insights on their function. Results obtained from this robust reductionist system can complement more time consuming and complex studies of such pathways in primary T cells.

Author Information

Sabrina Jutz

Annika Hennig

Wolfgang Paster

Ömer Asrak

Dejana Dijanovic

Florian Kellner

Winfried F. Pickl

Johannes B. Huppa

Judith Leitner

Peter Steinberger
Primary Contact  _


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