Abstract
In the thymus, low-affinity T cell antigen receptor (TCR) engagement facilitates positive selection of a useful T cell repertoire. Here we report that TCR responsiveness of mature CD8+ T cells is fine tuned by their affinity for positively selecting peptides in the thymus and that optimal TCR responsiveness requires positive selection on major histocompatibility complex class I–associated peptides produced by the thymoproteasome, which is specifically expressed in the thymic cortical epithelium. Thymoproteasome-independent positive selection of monoclonal CD8+ T cells results in aberrant TCR responsiveness, homeostatic maintenance and immune responses to infection. These results demonstrate a novel aspect of positive selection, in which TCR affinity for positively selecting peptides produced by thymic epithelium determines the subsequent antigen responsiveness of mature CD8+ T cells in the periphery.
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References
von Boehmer, H. The developmental biology of T lymphocytes. Annu. Rev. Immunol. 6, 309–326 (1988).
Palmer, E. Negative selection—clearing out the bad apples from the T-cell repertoire. Nat. Rev. Immunol. 3, 383–391 (2003).
Starr, T.K., Jameson, S.C. & Hogquist, K.A. Positive and negative selection of T cells. Annu. Rev. Immunol. 21, 139–176 (2003).
Bevan, M.J. Thymic education. Immunol. Today 2, 216–219 (1981).
Allen, P.M. Peptides in positive and negative selection: a delicate balance. Cell 76, 593–596 (1994).
Anderson, G., Owen, J.J., Moore, N.C. & Jenkinson, E.J. Thymic epithelial cells provide unique signals for positive selection of CD4+CD8+ thymocytes in vitro. J. Exp. Med. 179, 2027–2031 (1994).
Laufer, T.M., DeKoning, J., Markowitz, J.S., Lo, D. & Glimcher, L.H. Unopposed positive selection and autoreactivity in mice expressing class II MHC only on thymic cortex. Nature 383, 81–85 (1996).
Capone, M., Romagnoli, P., Beermann, F., MacDonald, H.R. & van Meerwijk, J.P. Dissociation of thymic positive and negative selection in transgenic mice expressing major histocompatibility complex class I molecules exclusively on thymic cortical epithelial cells. Blood 97, 1336–1342 (2001).
Klein, L., Hinterberger, M., Wirnsberger, G. & Kyewski, B. Antigen presentation in the thymus for positive selection and central tolerance induction. Nat. Rev. Immunol. 9, 833–844 (2009).
Takada, K. & Takahama, Y. Positive-selection-inducing self-peptides displayed by cortical thymic epithelial cells. Adv. Immunol. 125, 87–110 (2015).
Murata, S. et al. Regulation of CD8+ T cell development by thymus-specific proteasomes. Science 316, 1349–1353 (2007).
Nitta, T. et al. Thymoproteasome shapes immunocompetent repertoire of CD8+ T cells. Immunity 32, 29–40 (2010).
Takahama, Y., Takada, K., Murata, S. & Tanaka, K. β5t-containing thymoproteasome: specific expression in thymic cortical epithelial cells and role in positive selection of CD8+ T cells. Curr. Opin. Immunol. 24, 92–98 (2012).
Rock, K.L. & Goldberg, A.L. Degradation of cell proteins and the generation of MHC class I-presented peptides. Annu. Rev. Immunol. 17, 739–779 (1999).
Hogquist, K.A. et al. T cell receptor antagonist peptides induce positive selection. Cell 76, 17–27 (1994).
Brändle, D. et al. Involvement of both T cell receptor Vα and Vβ variable region domains and α chain junctional region in viral antigen recognition. Eur. J. Immunol. 21, 2195–2202 (1991).
Mamalaki, C. et al. Positive and negative selection in transgenic mice expressing a T-cell receptor specific for influenza nucleoprotein and endogenous superantigen. Dev. Immunol. 3, 159–174 (1993).
Stefanová, I., Dorfman, J.R. & Germain, R.N. Self-recognition promotes the foreign antigen sensitivity of naive T lymphocytes. Nature 420, 429–434 (2002).
Mandl, J.N., Monteiro, J.P., Vrisekoop, N. & Germain, R.N. T cell-positive selection uses self-ligand binding strength to optimize repertoire recognition of foreign antigens. Immunity 38, 263–274 (2013).
Xing, Y., Jameson, S.C. & Hogquist, K.A. Thymoproteasome subunit-β5T generates peptide-MHC complexes specialized for positive selection. Proc. Natl. Acad. Sci. USA 110, 6979–6984 (2013).
Moon, J.J. et al. Naive CD4+ T cell frequency varies for different epitopes and predicts repertoire diversity and response magnitude. Immunity 27, 203–213 (2007).
Moon, J.J. et al. Tracking epitope-specific T cells. Nat. Protoc. 4, 565–581 (2009).
Haluszczak, C. et al. The antigen-specific CD8+ T cell repertoire in unimmunized mice includes memory phenotype cells bearing markers of homeostatic expansion. J. Exp. Med. 206, 435–448 (2009).
Sosinowski, T. et al. CD8α+ dendritic cell trans presentation of IL-15 to naive CD8+ T cells produces antigen-inexperienced T cells in the periphery with memory phenotype and function. J. Immunol. 190, 1936–1947 (2013).
Obar, J.J. et al. Pathogen-induced inflammatory environment controls effector and memory CD8+ T cell differentiation. J. Immunol. 187, 4967–4978 (2011).
Van Laethem, F. et al. Lck availability during thymic selection determines the recognition specificity of the T cell repertoire. Cell 154, 1326–1341 (2013).
Yamashita, I., Nagata, T., Tada, T. & Nakayama, T. CD69 cell surface expression identifies developing thymocytes which audition for T cell antigen receptor-mediated positive selection. Int. Immunol. 5, 1139–1150 (1993).
Ueno, T. et al. CCR7 signals are essential for cortex-medulla migration of developing thymocytes. J. Exp. Med. 200, 493–505 (2004).
Azzam, H.S. et al. CD5 expression is developmentally regulated by T cell receptor (TCR) signals and TCR avidity. J. Exp. Med. 188, 2301–2311 (1998).
Love, P.E., Lee, J. & Shores, E.W. Critical relationship between TCR signaling potential and TCR affinity during thymocyte selection. J. Immunol. 165, 3080–3087 (2000).
Hogquist, K.A. et al. Identification of a naturally occurring ligand for thymic positive selection. Immunity 6, 389–399 (1997).
Santori, F.R. et al. Rare, structurally homologous self-peptides promote thymocyte positive selection. Immunity 17, 131–142 (2002).
Juang, J. et al. Peptide-MHC heterodimers show that thymic positive selection requires a more restricted set of self-peptides than negative selection. J. Exp. Med. 207, 1223–1234 (2010).
Fulton, R.B. et al. The TCR's sensitivity to self peptide-MHC dictates the ability of naive CD8+ T cells to respond to foreign antigens. Nat. Immunol. 16, 107–117 (2015).
Persaud, S.P., Parker, C.R., Lo, W.L., Weber, K.S. & Allen, P.M. Intrinsic CD4+ T cell sensitivity and response to a pathogen are set and sustained by avidity for thymic and peripheral complexes of self peptide and MHC. Nat. Immunol. 15, 266–274 (2014).
Sasaki, K. et al. Thymoproteasomes produce unique peptide motifs for positive selection of CD8+ T cells. Nat. Commun. 6, 7484 (2015).
Fu, G. et al. Fine-tuning T cell receptor signaling to control T cell development. Trends Immunol. 35, 311–318 (2014).
Hogquist, K.A. & Jameson, S.C. The self-obsession of T cells: how TCR signaling thresholds affect fate 'decisions' and effector function. Nat. Immunol. 15, 815–823 (2014).
Lee, J.Y., Hamilton, S.E., Akue, A.D., Hogquist, K.A. & Jameson, S.C. Virtual memory CD8 T cells display unique functional properties. Proc. Natl. Acad. Sci. USA 110, 13498–13503 (2013).
Brodin, P., Kärre, K. & Höglund, P. NK cell education: not an on-off switch but a tunable rheostat. Trends Immunol. 30, 143–149 (2009).
Brodin, P., Lakshmikanth, T., Johansson, S., Kärre, K. & Höglund, P. The strength of inhibitory input during education quantitatively tunes the functional responsiveness of individual natural killer cells. Blood 113, 2434–2441 (2009).
Joncker, N.T., Fernandez, N.C., Treiner, E., Vivier, E. & Raulet, D.H. NK cell responsiveness is tuned commensurate with the number of inhibitory receptors for self-MHC class I: the rheostat model. J. Immunol. 182, 4572–4580 (2009).
Shinkai, Y. et al. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell 68, 855–867 (1992).
Van Kaer, L., Ashton-Rickardt, P.G., Ploegh, H.L. & Tonegawa, S. TAP1 mutant mice are deficient in antigen presentation, surface class I molecules, and CD4−8+ T cells. Cell 71, 1205–1214 (1992).
Morse, H.C. III, Shen, F.W. & Hämmerling, U. Genetic nomenclature for loci controlling mouse lymphocyte antigens. Immunogenetics 25, 71–78 (1987).
Haring, J.S., Corbin, G.A. & Harty, J.T. Dynamic regulation of IFN-γ signaling in antigen-specific CD8+ T cells responding to infection. J. Immunol. 174, 6791–6802 (2005).
Acknowledgements
We thank H. Kosako, I. Ohigashi, M. Kozai and B. Kim for helpful discussion and reading the manuscript. This work was supported by grants from MEXT-JSPS (24111004 and 23249025 to Y.T. and 24790475 and 26460576 to K. Takada), Naito Foundation (to Y.T. and K. Takada), Takeda Science Foundation (to K. Takada), and Uehara Memorial Foundation (to K. Takada).
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K. Takada and Y.T. conceived the study. K. Takada, S.C.J., A.S. and Y.T. designed the experiments. K. Takada, F.V.L., Y.X., K.A., H.S., S.M. and K. Tanaka performed the experiments. K. Takada, S.C.J., A.S. and Y.T. wrote the manuscript.
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Supplementary Figure 1 TCR responsiveness of monoclonal T cells generated in the β5t-deficient thymus.
CD44loCD8+ OT-I-TCR-transgenic T cells generated in control or β5t-deficient mice were stimulated with OVAp–H-2Kb tetramers. Numbers in histograms indicate mean fluorescence intensity (MFI) of CD69 in CD69+ cells. Representative data from four independent experiments are shown. Bars indicate average ± standard errors of the mean. *P < 0.05.
Supplementary Figure 2 Impaired Erk phosphorylation in T cells generated in the β5t-deficient thymus.
ERK1 and 2 phosphorylation in response to OVAp–H-2Kb tetramer stimulation was examined in CD44loCD8+ OT-I-TCR-transgenic T cells at indicated time. Representative data from two independent experiments are shown.
Supplementary Figure 3 Peripheral T cells generated in a β5t-deficient thymus are abnormal.
(a) Frequencies of CD44loCD122lo and CD44hiCD122hi cells in CD8+ T cells from the spleens and lymph nodes of β5t-deficient Rag2-deficient OT-I-TCR-transgenic mice. (b) CD44 and CD122 expression in CD4−CD8+Vα2hi mature OT-I-TCR-transgenic thymocytes. (c) Splenic CD8+ T cells in β5t-deficient mice at different ages were analyzed for the ratios of CD44lo to CD44hi cells. (d) Absolute numbers of naïve (CD44loCD122lo) and memory-like (CD44hiCD122hi) CD8+ T cells in polyclonal TCR-expressing β5t-deficient mice at different ages. (e) Thymocytes from β5t-deficient and control mice were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and adoptively transferred to sublethally irradiated (6.0 Gy) B6-Ly5.1 mice (1×106 SP thymocytes/recipient). CD45.2+ donor cells in recipient spleens were analyzed for CFSE dilution as well as CD4, CD8, and CD44 expression 14 days after transfer. (f) Representative histograms of CD8+ T cells bound to gp33–H-2Db tetramers in the experiments shown in Fig. 3c. Accumulated results from more than six (a and b) or three (c and d) mice/group and representative data from two (e) or three (f) independent experiments are shown. Graphs show data of individual mice (circles) and means (bars) (a and b). Circles and bars indicate average ± standard errors of the mean (c-e). ***P < 0.001, **P < 0.01 *P < 0.05.
Supplementary Figure 4 Increased frequency of memory-like cells in CD8+ T cell compartment.
CD44 and CD122 expression in T cells from the spleens and lymph nodes of β5t-deficient mice were analyzed. Graphs show data of individual mice (circles) and means (bars). Accumulated results from 5-6 mice per group. *P < 0.001.
Supplementary Figure 5 β5t-dependent positive selection affects monoclonal T cell responses to infection.
(a) Graphical scheme of LM-OVA infection experiments. β5t-dependent or -independent OT-I-TCR-transgenic T cells were obtained from bone marrow chimeras (Rag1−/− OT-I > Psmb11+/+ or Rag1−/− OT-I > Psmb11−/−). Bone marrow chimera-derived OT-I-TCR-transgenic T cells (CD45.2+CD90.2+) were co-transferred with normal OT-I T cells (CD45.2+CD90.1+) into B6-Ly5.1 mice (CD45.1+CD45.2−). Blood lymphocytes were analyzed 5 and 8 days following LM-OVA infection. (b) Expression of CD44 and KLRG1 in donor OT-I T cells was examined 8 days after infection in the experiments shown in Fig. 4. Graphs show data of individual mice (circles) and means (bars). Cumulative data from two independent experiments. *P < 0.001.
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Takada, K., Van Laethem, F., Xing, Y. et al. TCR affinity for thymoproteasome-dependent positively selecting peptides conditions antigen responsiveness in CD8+ T cells. Nat Immunol 16, 1069–1076 (2015). https://doi.org/10.1038/ni.3237
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DOI: https://doi.org/10.1038/ni.3237
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