Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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

Abstract

Interactions of T cell antigen receptors (TCRs) with complexes of self peptide and major histocompatibility complex (MHC) are crucial to T cell development, but their role in peripheral T cell responses remains unclear. Specific and nonspecific stimulation of LLO56 and LLO118 T cells, which transgenically express a TCR specific for the same Listeria monocytogenes epitope, elicited distinct interleukin 2 (IL-2) and phosphorylated kinase Erk responses, the strength of which was set in the thymus and maintained in the periphery in proportion to the avidity of the binding of the TCR to the self peptide–MHC complex. Deprivation of self peptide–MHC substantially compromised the population expansion of LLO56 T cells in response to L. monocytogenes in vivo. Despite their very different self-reactivity, LLO56 T cells and LLO118 T cells bound cognate peptide–MHC with an identical affinity, which challenges associations made between these parameters. Our findings highlight a crucial role for selecting ligands encountered during thymic 'education' in determining the intrinsic functionality of CD4+ T cells.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: LLO56 T cells and LLO118 T cells diverge in their IL-2 responses to specific or nonspecific stimuli.
Figure 2: The stronger IL-2 responses of LLO56 T cells are linked to greater activation-induced phosphorylation of Erk and basal phosphorylation of TCRζ than that of LLO118 T cells.
Figure 3: Strength of intrinsic IL-2 responses and signaling in polyclonal B6 CD4+ and CD8+ T cells correlates with CD5 expression.
Figure 4: Functional attributes of LLO56 and LLO118 T cells emerge during positive selection, during which the LLO56 TCR receives a stronger signal from selecting self peptide–MHC.
Figure 5: Deprivation of self peptide–MHC compromises intrinsic IL-2 and Erk responses and in vivo responses to L. monocytogenes.
Figure 6: CD5 antagonizes signaling from self peptide–MHC and intrinsic IL-2 and phosphorylated Erk responses.

References

  1. van der Merwe, P.A. & Davis, S.J. Molecular interactions mediating T cell antigen recognition. Annu. Rev. Immunol. 21, 659–684 (2003).

    CAS  PubMed  Google Scholar 

  2. Davis, M.M. et al. Ligand recognition by αβ T cell receptors. Annu. Rev. Immunol. 16, 523–544 (1998).

    CAS  PubMed  Google Scholar 

  3. Morris, G.P. & Allen, P.M. How the TCR balances sensitivity and specificity for the recognition of self and pathogens. Nat. Immunol. 13, 121–128 (2012).

    CAS  PubMed  Google Scholar 

  4. Ernst, B., Lee, D.S., Chang, J.M., Sprent, J. & Surh, C.D. The peptide ligands mediating positive selection in the thymus control T cell survival and homeostatic proliferation in the periphery. Immunity 11, 173–181 (1999).

    CAS  PubMed  Google Scholar 

  5. Krogsgaard, M. et al. Agonist/endogenous peptide-MHC heterodimers drive T cell activation and sensitivity. Nature 434, 238–243 (2005).

    CAS  PubMed  Google Scholar 

  6. Lo, W.L. et al. An endogenous peptide positively selects and augments the activation and survival of peripheral CD4+ T cells. Nat. Immunol. 10, 1155–1161 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Kirberg, J., Berns, A. & von Boehmer, H. Peripheral T cell survival requires continual ligation of the T cell receptor to major histocompatibility complex-encoded molecules. J. Exp. Med. 186, 1269–1275 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Cho, J.H., Kim, H.O., Surh, C.D. & Sprent, J. T cell receptor-dependent regulation of lipid rafts controls naive CD8+ T cell homeostasis. Immunity 32, 214–226 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Kersh, G.J. et al. Structural and functional consequences of altering a peptide MHC anchor residue. J. Immunol. 166, 3345–3354 (2001).

    CAS  PubMed  Google Scholar 

  10. Hataye, J., Moon, J.J., Khoruts, A., Reilly, C. & Jenkins, M.K. Naive and memory CD4+ T cell survival controlled by clonal abundance. Science 312, 114–116 (2006).

    CAS  PubMed  Google Scholar 

  11. Obar, J.J., Khanna, K.M. & Lefrancois, L. Endogenous naive CD8+ T cell precursor frequency regulates primary and memory responses to infection. Immunity 28, 859–869 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Malherbe, L., Hausl, C., Teyton, L. & McHeyzer-Williams, M.G. Clonal selection of helper T cells is determined by an affinity threshold with no further skewing of TCR binding properties. Immunity 21, 669–679 (2004).

    CAS  PubMed  Google Scholar 

  13. Busch, D.H. & Pamer, E.G. T cell affinity maturation by selective expansion during infection. J. Exp. Med. 189, 701–710 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Zehn, D., Lee, S.Y. & Bevan, M.J. Complete but curtailed T-cell response to very low-affinity antigen. Nature 458, 211–214 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Weber, K.S. et al. Distinct CD4+ helper T cells involved in primary and secondary responses to infection. Proc. Natl. Acad. Sci. USA 109, 9511–9516 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Lenardo, M. et al. Mature T lymphocyte apoptosis–immune regulation in a dynamic and unpredictable antigenic environment. Annu. Rev. Immunol. 17, 221–253 (1999).

    CAS  PubMed  Google Scholar 

  20. Hochweller, K. et al. Dendritic cells control T cell tonic signaling required for responsiveness to foreign antigen. Proc. Natl. Acad. Sci. USA 107, 5931–5936 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Stefanová, I., Dorfman, J.R. & Germain, R.N. Self-recognition promotes the foreign antigen sensitivity of naive T lymphocytes. Nature 420, 429–434 (2002).

    PubMed  Google Scholar 

  22. Lo, W.L., Donermeyer, D.L. & Allen, P.M. A voltage-gated sodium channel is essential for the positive selection of CD4+ T cells. Nat. Immunol. 13, 880–887 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Kaye, J., Kersh, G., Engel, I. & Hedrick, S.M. Structure and specificity of the T cell antigen receptor. Semin. Immunol. 3, 269–281 (1991).

    CAS  PubMed  Google Scholar 

  24. Moran, A.E. et al. T cell receptor signal strength in Treg and iNKT cell development demonstrated by a novel fluorescent reporter mouse. J. Exp. Med. 208, 1279–1289 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Smith, K. et al. Sensory adaptation in naive peripheral CD4 T cells. J. Exp. Med. 194, 1253–1261 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Skoberne, M., Holtappels, R., Hof, H. & Geginat, G. Dynamic antigen presentation patterns of Listeria monocytogenes-derived CD8 T cell epitopes in vivo. J. Immunol. 167, 2209–2218 (2001).

    CAS  PubMed  Google Scholar 

  27. Tarakhovsky, A. et al. A role for CD5 in TCR-mediated signal transduction and thymocyte selection. Science 269, 535–537 (1995).

    CAS  PubMed  Google Scholar 

  28. Zhou, X.Y. et al. CD5 costimulation up-regulates the signaling to extracellular signal-regulated kinase activation in CD4+CD8+ thymocytes and supports their differentiation to the CD4 lineage. J. Immunol. 164, 1260–1268 (2000).

    CAS  PubMed  Google Scholar 

  29. Peña-Rossi, C. et al. Negative regulation of CD4 lineage development and responses by CD5. J. Immunol. 163, 6494–6501 (1999).

    PubMed  Google Scholar 

  30. Azzam, H.S. et al. Fine tuning of TCR signaling by CD5. J. Immunol. 166, 5464–5472 (2001).

    CAS  PubMed  Google Scholar 

  31. Gapin, L., Matsuda, J.L., Surh, C.D. & Kronenberg, M. NKT cells derive from double-positive thymocytes that are positively selected by CD1d. Nat. Immunol. 2, 971–978 (2001).

    CAS  PubMed  Google Scholar 

  32. Lio, C.W. & Hsieh, C.S. A two-step process for thymic regulatory T cell development. Immunity 28, 100–111 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Stritesky, G.L., Jameson, S.C. & Hogquist, K.A. Selection of self-reactive T cells in the thymus. Annu. Rev. Immunol. 30, 95–114 (2012).

    CAS  PubMed  Google Scholar 

  34. Zikherman, J., Parameswaran, R. & Weiss, A. Endogenous antigen tunes the responsiveness of naive B cells but not T cells. Nature 489, 160–164 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Daniels, M.A. et al. Thymic selection threshold defined by compartmentalization of Ras/MAPK signalling. Nature 444, 724–729 (2006).

    CAS  PubMed  Google Scholar 

  36. Huang, J. et al. The kinetics of two-dimensional TCR and pMHC interactions determine T-cell responsiveness. Nature 464, 932–936 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Sabatino, J.J. Jr., Huang, J., Zhu, C. & Evavold, B.D. High prevalence of low affinity peptide-MHC II tetramer-negative effectors during polyclonal CD4+ T cell responses. J. Exp. Med. 208, 81–90 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Tubo, N.J. et al. Single naive CD4+ T cells from a diverse repertoire produce different effector cell types during infection. Cell 153, 785–796 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Kim, C., Wilson, T., Fischer, K.F. & Williams, M.A. Sustained interactions between T cell receptors and antigens promote the differentiation of CD4+ memory T cells. Immunity 39, 508–520 (2013).

    CAS  PubMed  Google Scholar 

  40. Martin, B. et al. Highly self-reactive naive CD4 T cells are prone to differentiate into regulatory T cells. Nat. Commun. 4, 2209 (2013).

    PubMed  Google Scholar 

  41. Kersh, E.N., Shaw, A.S. & Allen, P.M. Fidelity of T cell activation through multistep T cell receptor zeta phosphorylation. Science 281, 572–575 (1998).

    CAS  PubMed  Google Scholar 

  42. Persaud, S.P., Donermeyer, D.L., Weber, K.S., Kranz, D.M. & Allen, P.M. High-affinity T cell receptor differentiates cognate peptide-MHC and altered peptide ligands with distinct kinetics and thermodynamics. Mol. Immunol. 47, 1793–1801 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Weber, K.S., Donermeyer, D.L., Allen, P.M. & Kranz, D.M. Class II-restricted T cell receptor engineered in vitro for higher affinity retains peptide specificity and function. Proc. Natl. Acad. Sci. USA 102, 19033–19038 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Garcia, K.C., Radu, C.G., Ho, J., Ober, R.J. & Ward, E.S. Kinetics and thermodynamics of T cell receptor- autoantigen interactions in murine experimental autoimmune encephalomyelitis. Proc. Natl. Acad. Sci. USA 98, 6818–6823 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Hallaq, H. et al. Activation of protein kinase C alters the intracellular distribution and mobility of cardiac Na+ channels. Am. J. Physiol. Heart Circ. Physiol. 302, H782–H789 (2012).

    CAS  PubMed  Google Scholar 

  46. Moon, J.J. et al. Tracking epitope-specific T cells. Nat. Protoc. 4, 565–581 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank E. Huseby (University of Massachusetts Medical School) for soluble I-Ab; Q.J. Li for generating the LLO56 and LLO118 TCR-encoding transgene constructs; J. Ting (University of North Carolina Chapel Hill School of Medicine) for mice doubly deficient in H-2M and β2-microglobulin; K. Murray (Vanderbilt University School of Medicine) for the SCN5A–green fluorescent protein construct; D. Kreamalmeyer for mouse breeding and care; S. Horvath for peptide synthesis; D. Brinja and E. Lantelme for assistance with sorting by flow cytometry; D. Donermeyer, A. Shaw, E. Unanue and E. Brown for comments on the manuscript; and A. Chakraborty, M. Davis, M. Dustin, M. Kardar, E. Pamer, A. Perelson, D. Portnoy and A. Shaw (members of the program project (AI-071195) under which this work was initiated). Supported by the US National Institutes of Health (AI-24157).

Author information

Authors and Affiliations

Authors

Contributions

S.P.P., K.S.W. and P.M.A. designed the study; S.P.P., K.S.W., C.R.P. and W.-L.L. did all experiments; S.P.P. and P.M.A. wrote the manuscript; K.S.W. did the initial studies with LLO56 and LLO118 mice; and all authors read, commented on and approved of the manuscript before submission.

Corresponding author

Correspondence to Paul M Allen.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Phenotyping and in vivo proliferative responses of mature LLO56 and LLO118 T cells.

(a) In vivo proliferation of CFSE-labeled LLO56 and LLO118 T cells in response to Listeria at 2, 4, and 6 days post-infection. The presented FACS plots are representative of 6-7 mice analyzed over four experiments. (b) Expression of signaling and costimulatory molecules on LLO56 and LLO118 T cells. The distribution of CD5 expression for polyclonal B6 CD4+ T cells is included in the CD5 histogram for reference. Data are representative of at least three mice from at least three experiments.

Supplementary Figure 2 Validation of LLO56 and LLO118 signaling results.

(a) Immunoblot analysis of ERK phosphorylation kinetics of PMA-stimulated LLO56 and LLO118 CD4+ T cells, representative of two experiments. (b) Confirmation of p21 band in 4G10 blots of unstimulated CD4+ whole cell lysates as phospho-TCRζ, using rabbit polyclonal anti-ζ serum 777. 4G10 and anti-ζ serum staining was performed on the same blot. LLO56 was used for this validation as it gave the most easily detectable p21 band. Data are representative of at least three experiments.

Supplementary Figure 3 CD5hi B6 CD4+ and CD8+ T cells are more responsive to stimulation than are their CD5lo counterparts.

(a) Primary (top) and graphed (bottom) data from IL-2 capture assay analysis of CD4+ and CD8+ T cells stimulated with αCD3 + αCD28. Cells were gated into four equal fractions based on CD5 expression (Q1 through Q4, from lowest to highest CD5 expression). (b) Comparison of CD5 expression on stimulated and unstimulated CD4+ and CD8+ T cells in IL-2 capture (left), intracellular IL-2 (middle), and ERK phosphorylation (right) assays. (c) Overlays of B6 CD4+ and CD8+ T cells pre-sorted into CD5 fractions (Q1 through Q4) by flow cytometry, then analyzed for PMA + ionomycin-induced IL-2 (top) and PMA-induced phospho-ERK responses (3 minute stimulation, bottom). Data are representative of two or three experiments. Bar graphs depict means ± SEM, with statistical analyses done using unpaired two-tailed Student's t tests. *P < 0.01 and **P < 0.001.

Source data

Supplementary Figure 4 Ectopic expression of the SCN4B and SCN5A voltage-gated sodium channel subunits confers self-reactivity to CD4+ T cells in proportion to CD5 expression.

(a) Gating strategy for the experiments, showing identification of transfected VGSC+ T cells. (b) CD69 upregulation response of VGSC+ LLO56, LLO118 and B6 CD4+ T cells when cultured with or without B6 APCs, or with APCs pretreated with anti I-Ab. Representative primary data (left) and compiled data (right) are presented. (c) Comparison of CD69 responses of untransfected (SCN5A-SCN4B-), singly-transfected (SCN5A-SCN4B+), and doubly-transfected VGSC+ T cells (SCN5A+SCN4B+), in the presence of B6 APCs (without I-Ab blockade). Bars depict means ± SEM. For cells cultured with and without APCs, results from eight cultures over three experiments (LLO56 and LLO118), or six cultures over two experiments (B6 CD4+) were compiled for the graph in panel (b); for blocking studies with anti-I-Ab, 3 or 4 cultures over two experiments were compiled. Statistical analysis was done using a two-tailed Mann-Whitney test. *P < 0.001.

Source data

Supplementary Figure 5 Detailed analysis of selection and activation responses in thymocytes with transgenic expression of a TCR.

(a) Identification of post-selection (TCRhiCD69+ thymocytes) from total viable LLO56 and LLO118 thymocytes, representative of at least three experiments. (b) Frequencies of NK (CD3-NK1.1+), NKT (CD3+NK1.1+), and γδ T cells (GL3+) among DN thymocytes. Data are representative of analyses of three mice from two experiments. (c) Gradual emergence of LLO56 and LLO118 responses during the DP to CD4SP thymocyte transition (top plots, labeled 1 through 5; DN thymocytes gated out for clarity). PMA + ionomycin-induced IL-2 responses (bottom left, numbers indicate % IL-2+ cells) and ERK phosphorylation after 3 minute PMA stimulation (bottom right, red and blue numbers indicate LLO56 and LLO118 pERK MFI, respectively) are presented. Data are representative of at least three experiments. (d) PMA + ionomycin-induced intracellular IL-2 responses of CD4+ T cells transgenically expressing the AND TCR on H2b or H2k MHC haplotype backgrounds representative of three experiments. (e) Analysis of cell survival-associated markers among LLO56 and LLO118 thymocyte subsets and mature CD4+ T cells, representative of two or three experiments. Bar graphs depict means ± SEM, with statistical analyses done using unpaired two-tailed Student's t tests. *P < 0.0001.

Source data

Supplementary Figure 6 Loss of the IL-2 responses of LLO56 and LLO118 T cells in adoptive-transfer experiments tracks with deprivation of self peptide–MHC class II.

(a) Cell surface phenotype of LLO56 and LLO118 T cells following 4-day transfer to B6 or MHC II-deficient recipients, representative of at least three experiments. (b) Ex vivo analysis of PMA + ionomycin-induced IL-2 responses of LLO56 and LLO118 T cells following 4-day transfer to TCR Cα-deficient (LLO56 n = 8, LLO118 n = 6) or H-2M-deficient (LLO56 n = 11, LLO118 n = 10) recipients. Data are compilations of four experiments. (c) Ex vivo analysis of PMA + ionomycin-induced IL-2 responses of freshly isolated LLO56 T cells (n = 4) or LLO56 T cells transferred to H-2M-deficient recipients for the indicated periods of time (Day 1 n = 8, Day 2 n = 6, Day 4 n = 11). (d) Schematic of experiment testing effect of self-pMHC withdrawal on LLO56 and LLO118 T cell response to Listeria in vivo. For (b) and (c), each data point comprising the bar graphs is the % IL-2+ LLO T cells from a single recipient. Bar graphs depict means ± SEM, with statistical analyses done using unpaired two-tailed Student's t tests. NS, not significant and *P < 0.0001.

Source data

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 (PDF 6153 kb)

Source data

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Persaud, S., Parker, C., Lo, WL. et al. 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). https://doi.org/10.1038/ni.2822

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.2822

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing