TOX reinforces the phenotype and longevity of exhausted T cells in chronic viral infection

Francesca Alfei, Kristiyan Kanev, Maike Hofmann, Ming Wu, Hazem E. Ghoneim, Patrick Roelli, Daniel T. Utzschneider, Madlaina von Hoesslin, Jolie G. Cullen, Yiping Fan, Vasyl Eisenberg, Dirk Wohlleber, Katja Steiger, Doron Merkler, Mauro Delorenzi, Percy A. Knolle, Cyrille J. Cohen, Robert Thimme, Benjamin Youngblood, Dietmar Zehn

Research output: Contribution to journalArticlepeer-review

398 Scopus citations


Cytotoxic T cells are essential mediators of protective immunity to viral infection and malignant tumours and are a key target of immunotherapy approaches. However, prolonged exposure to cognate antigens often attenuates the effector capacity of T cells and limits their therapeutic potential1–4. This process, known as T cell exhaustion or dysfunction1, is manifested by epigenetically enforced changes in gene regulation that reduce the expression of cytokines and effector molecules and upregulate the expression of inhibitory receptors such as programmed cell-death 1 (PD-1)5–8. The underlying molecular mechanisms that induce and stabilize the phenotypic and functional features of exhausted T cells remain poorly understood9–12. Here we report that the development and maintenance of populations of exhausted T cells in mice requires the thymocyte selection-associated high mobility group box (TOX) protein13–15. TOX is induced by high antigen stimulation of the T cell receptor and correlates with the presence of an exhausted phenotype during chronic infections with lymphocytic choriomeningitis virus in mice and hepatitis C virus in humans. Removal of its DNA-binding domain reduces the expression of PD-1 at the mRNA and protein level, augments the production of cytokines and results in a more polyfunctional T cell phenotype. T cells with this deletion initially mediate increased effector function and cause more severe immunopathology, but ultimately undergo a massive decline in their quantity, notably among the subset of TCF-1+ self-renewing T cells. Altogether, we show that TOX is a critical factor for the normal progression of T cell dysfunction and the maintenance of exhausted T cells during chronic infection, and provide a link between the suppression of effector function intrinsic to CD8 T cells and protection against immunopathology.

Original languageEnglish
Pages (from-to)265-269
Number of pages5
Issue number7764
StatePublished - 11 Jul 2019

Bibliographical note

Funding Information:
Mice. P14 TCRαβ-transgenic mice were provided by A. Oxenius27 and Vβ5 TCRβ-only transgenic mice28 by P. Fink. The C57BL/6N Toxtm1a(KOMP)Wtsi mouse strain used for this project was generated by the trans-NIH Knockout Mouse Project (KOMP) and obtained from the KOMP Repository ( NIH grants to Velocigene at Regeneron (U01HG004085) and the CSD Consortium (U01HG004080) funded the generation of gene-targeted embryonic stem cells for 8,500 genes in the KOMP Program and archived and distributed by the KOMP Repository at UC Davis and CHORI (U42RR024244). For more information or to obtain KOMP products go to or email service@komp. org. The C57BL/6N Toxtm1a(KOMP)Wtsi line was crossed with a FLP-deleter strain to eliminate the LacZ reporter construct and to convert them into Toxtm1c(KOMP)Wtsi mice. The progeny of these mice were crossed with Mx1cre (also known as Mxcre) or Cd4cre, Rosa26-STOP-eYFP (Jackson Laboratories) and P14 transgenic mice, which were subsequently crossed to generate Toxtm1c(KOMP)Wtsi;Mxcre;Rosa26-STOP-eYFP P14 or Toxtm1c(KOMP)Wtsi;CD4cre;Rosa26-stop-eYFP P14 quadruple transgenic mice. In addition, Toxtm1c(KOMP)Wtsi;GzmbcreERT2 were generated. Mxcre;Rosa26-STOP-eYFP P14, Cd4cre;Rosa26-STOP-eYFP P14, or GzmbcreERT2 mice were used as genetically matched controls.

Funding Information:
Acknowledgements We thank M. J. Bevan and M. Prlic for input, feedback and suggestions; T. Herbinger and B. Dötterböck; W. Schmid and C. Amette for technical assistance; and S. Schleicher and C. Lechner for animal husbandry. Work in the D.Z. laboratory was supported by a ‘European Research Council starting grant’ (ProtecTC) and subsequently a ‘European Research Council consolidator grant’ (ToCCaTa), grants from the Swiss National Science Foundation (CRSII3_160708, 310030E-164187, 51PHP0_157319 and PP00P3_144883), the Swiss Vaccine Research Institute (SVRI) and grants from the German Research Foundation (SFB1054). D.Z. and C.J.C. are supported by a German-Israeli Foundation for Scientific Research and Development (GIF) grant (I-1440-414.13/2017). K.S. is supported by German Research Foundation grants (SFB824 and SFB1335). R.T. and M.H. are funded by a German Research Foundation grant (TRR179-TP01). B.Y. is supported by the NIH (R01AI114442) and the American Lebanese Syrian Associated Charities (ALSAC). P.A.K. is supported by the Germany Center for Infection Research Munich site and the CRC TRR179.

Publisher Copyright:
© 2019, The Author(s), under exclusive licence to Springer Nature Limited.


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