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The tyrosine phosphatase PTPN22 discriminates weak self peptides from strong agonist TCR signals

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Abstract

T cells must be tolerant of self antigens to avoid autoimmunity but responsive to foreign antigens to provide protection against infection. We found that in both naive T cells and effector T cells, the tyrosine phosphatase PTPN22 limited signaling via the T cell antigen receptor (TCR) by weak agonists and self antigens while not impeding responses to strong agonist antigens. T cells lacking PTPN22 showed enhanced formation of conjugates with antigen-presenting cells pulsed with weak peptides, which led to activation of the T cells and their production of inflammatory cytokines. This effect was exacerbated under conditions of lymphopenia, with the formation of potent memory T cells in the absence of PTPN22. Our data address how loss-of-function PTPN22 alleles can lead to the population expansion of effector and/or memory T cells and a predisposition to human autoimmunity.

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Figure 1: PTPN22 limits the basal activation of OT-I T cells.
Figure 2: Lymphopenia-induced proliferation is inhibited by PTPN22.
Figure 3: PTPN22 limits triggering of the TCR induced by weak agonists in naive T cells.
Figure 4: PTPN22 inhibits TCR-induced metabolic changes, transcription factor expression and proliferation.
Figure 5: PTPN22 restrains weak agonist–induced production of inflammatory cytokines by effector CTLs.
Figure 6: PTPN22 limits the proinflammatory capacity of memory T cells induced by homeostatic proliferation.
Figure 7: Enhanced LFA-1-dependent adhesion and conjugate formation by Ptpn22−/− T cells is important for their greater responses to weak antigens.

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Acknowledgements

We thank H. Shen (University of Pennsylvania) for the attenuated OVA-expressing L. monocytogenes strain with deletion of the actin assembly–inducing protein; D. Wright and C. Garcia for technical assistance, and P. Travers for reading the manuscript. Supported by the Wellcome Trust (096669 to R.Z., and 095831 for the Centre for Immunity, Infection and Evolution of the University of Edinburgh) and the Biotechnology and Biological Sciences Research Council (801148 to V.L.M.).

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Authors and Affiliations

Authors

Contributions

R.J.S. designed and did most in vitro experiments; R.J.B. designed and did most in vivo experiments and in vitro analyses of the formation of T cell conjugates; V.L.M. designed and did analyses of adhesion under shear flow; R.Z. designed experiments and led the overall project; and R.J.S. and R.Z. wrote the manuscript.

Corresponding author

Correspondence to Rose Zamoyska.

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Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Basal phenotype of OT-I thymus and lymph node T cells.

(a) Total cell numbers in wildtype (WT) and Ptpn22-/- (KO) thymi. (b) FACS dotplots showing distribution of thymocyte populations in 7 wk old WT and Ptpn22-/- OT-1 Rag1-/- mice. (c) Proportions of gated CD4+CD8+ double-positive (DP), and CD4+ and CD8+ single-positive (SP) thymocytes. (d) Absolute cell numbers of gated DP and SP thymocytes. (e) FACS histograms showing levels of expression of surface markers by gated CD8+ LN OT-1 T cells, as indicated. Data are representative of 8 mice of each genotype. In graphs, lines represent means and dots values from individual mice. NS - not significant, * p<0.001 by Student’s t-test.

Supplementary Figure 2 PTPN22 regulates early activation of T cells.

Wildtype (WT) and Ptpn22-/- (KO) OT-1 T cells were left unstimulated (US) or stimulated (stim) for 24h with 1 μM peptide (as indicated) and stained for cell surface and intracellular markers. Histograms showing the mean fluorescence intensities following intracellular staining for T-bet (a), c-Myc (b), eomesodermin (Eomes) (c) and IRF4 (d). (e) Cell size was assessed by analysis of forward scatter area (FSC-A) and levels of NB-d-glucose uptake (f) and surface expression of CD71 (g) and CD98 (h) measured. Data are representative of triplicate samples within 1 of at least 4 repeated experiments.

Supplementary Figure 3 Flow cytometry of CTLs generated in vitro.

(a) Comparable expansion of day 2 N4-stimulated wildtype (WT) and Ptpn22-/- OT-1 T cells in IL-2. Values represent means (n=3 replicate samples) and error bars SD. (b) Histograms show cell size (FSC-A) and expression of cell surface receptors, as indicated, by d6 IL-2-generated WT and Ptpn22-/- CTLs. (c) Intracellular expression of transcription factors and effector proteins by d6 IL-2-generated WT and Ptpn22-/- CTLs.

Supplementary Figure 4 Elevated inflammatory cytokine production by Ptpn22–/– (KO) CTLs.

(a) Histograms show levels of intracellular TNF and GM-CSF expressed by d6 WT and Ptpn22-/- CTLs following 4 h cultures unstimulated (US) or re-stimulation with 1 μM G4 peptide (stim). Data are representative of >3 repeated experiments. (b) Levels of MIP1α protein in culture supernatants following 24 h of G4 restimulation of d6 WT and Ptpn22-/- CTLs. Values represent means ± SD (n=3) from 1 of 2 repeated experiments.

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Salmond, R., Brownlie, R., Morrison, V. et al. The tyrosine phosphatase PTPN22 discriminates weak self peptides from strong agonist TCR signals. Nat Immunol 15, 875–883 (2014). https://doi.org/10.1038/ni.2958

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