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.

  • Article
  • Published:

Production of IL-10 by CD4+ regulatory T cells during the resolution of infection promotes the maturation of memory CD8+ T cells

Nature Immunology volume 16, pages 871–879 (2015)Cite this article

Subjects

Abstract

Memory CD8+ T cells are critical for host defense upon reexposure to intracellular pathogens. We found that interleukin 10 (IL-10) derived from CD4+ regulatory T cells (Treg cells) was necessary for the maturation of memory CD8+ T cells following acute infection with lymphocytic choriomeningitis virus (LCMV). Treg cell–derived IL-10 was most important during the resolution phase, calming inflammation and the activation state of dendritic cells. Adoptive transfer of IL-10-sufficient Treg cells during the resolution phase 'restored' the maturation of memory CD8+ T cells in IL-10-deficient mice. Our data indicate that Treg cell–derived IL-10 is needed to insulate CD8+ T cells from inflammatory signals, and reveal that the resolution phase of infection is a critical period that influences the quality and function of developing memory CD8+ T cells.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: IL-10 is required during the resolution phase of infection to allow optimal maturation of memory CD8+ T cells.
Figure 2: IL-10 is important in insulating CD8+ T cells from inflammatory signals following viral infection.
Figure 3: CD4+ Treg cell–derived IL-10 is critical for the maturation of memory CD8+ T cells.
Figure 4: CD4+ Treg cell–derived IL-10 is necessary for the formation of protective memory CD8+ T cells.
Figure 5: Virus-specific CD8+ T cells from mice lacking CD4+ Treg cell–derived IL-10 display a robust inflammatory gene signature.
Figure 6: Enhanced maturation of DCs and inflammation in the absence of Treg cell–derived IL-10.
Figure 7: Transfer of IL-10-competent CD4+ Treg cells during the resolution phase of LCMV infection is sufficient to 'rescue' the defect in the maturation of memory CD8+ T cells in Il10−/− mice.

Similar content being viewed by others

Accession codes

Primary accessions

Gene Expression Omnibus

References

  1. Ahmed, R. & Gray, D. Immunological memory and protective immunity: understanding their relation. Science 272, 54–60 (1996).

    CAS  PubMed  Google Scholar 

  2. Korber, B.T., Letvin, N.L. & Haynes, B.F. T-cell vaccine strategies for human immunodeficiency virus, the virus with a thousand faces. J. Virol. 83, 8300–8314 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Kaech, S.M. & Ahmed, R. Memory CD8+ T cell differentiation: initial antigen encounter triggers a developmental program in naïve cells. Nat. Immunol. 2, 415–422 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Kaech, S.M. & Cui, W. Transcriptional control of effector and memory CD8+ T cell differentiation. Nat. Rev. Immunol. 12, 749–761 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Cui, W., Liu, Y., Weinstein, J.S., Craft, J. & Kaech, S.M. An interleukin-21-interleukin-10-STAT3 pathway is critical for functional maturation of memory CD8+ T cells. Immunity 35, 792–805 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Rutishauser, R.L. et al. Transcriptional repressor Blimp-1 promotes CD8+ T cell terminal differentiation and represses the acquisition of central memory T cell properties. Immunity 31, 296–308 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Kim, M.V., Ouyang, W., Liao, W., Zhang, M.Q. & Li, M.O. The transcription factor Foxo1 controls central-memory CD8+ T cell responses to infection. Immunity 39, 286–297 (2013).

    CAS  PubMed  Google Scholar 

  8. Joshi, N.S. et al. Inflammation directs memory precursor and short-lived effector CD8+ T cell fates via the graded expression of T-bet transcription factor. Immunity 27, 281–295 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Yang, C.Y. et al. The transcriptional regulators Id2 and Id3 control the formation of distinct memory CD8+ T cell subsets. Nat. Immunol. 12, 1221–1229 (2011).

    CAS  PubMed  Google Scholar 

  10. Zhou, X. et al. Differentiation and persistence of memory CD8+ T cells depend on T cell factor 1. Immunity 33, 229–240 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Araki, K. et al. mTOR regulates memory CD8 T-cell differentiation. Nature 460, 108–112 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Rao, R.R., Li, Q., Gubbels Bupp, M.R. & Shrikant, P.A. Transcription factor Foxo1 represses T-bet-mediated effector functions and promotes memory CD8+ T cell differentiation. Immunity 36, 374–387 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Haring, J.S. & Harty, J.T. Interleukin-18-related genes are induced during the contraction phase but do not play major roles in regulating the dynamics or function of the T-cell response to Listeria monocytogenes infection. Infect. Immun. 77, 1894–1903 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Stelekati, E. et al. Bystander chronic infection negatively impacts development of CD8+ T cell memory. Immunity 40, 801–813 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Sun, J.C., Williams, M.A. & Bevan, M.J. CD4+ T cells are required for the maintenance, not programming, of memory CD8+ T cells after acute infection. Nat. Immunol. 5, 927–933 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Foulds, K.E., Rotte, M.J. & Seder, R.A. IL-10 is required for optimal CD8 T cell memory following Listeria monocytogenes infection. J. Immunol. 177, 2565–2574 (2006).

    CAS  PubMed  Google Scholar 

  17. Vignali, D.A.A., Collison, L.W. & Workman, C.J. How regulatory T cells work. Nat. Rev. Immunol. 8, 523–532 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Suvas, S., Kumaraguru, U., Pack, C.D., Lee, S. & Rouse, B.T. CD4+CD25+ T cells regulate virus-specific primary and memory CD8+ T cell responses. J. Exp. Med. 198, 889–901 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Pace, L. et al. Regulatory T cells increase the avidity of primary CD8+ T cell responses and promote memory. Science 338, 532–536 (2012).

    CAS  PubMed  Google Scholar 

  20. de Goër de Herve, M.G., Jaafoura, S., Vallée, M. & Taoufik, Y. FoxP3+ regulatory CD4 T cells control the generation of functional CD8 memory. Nat. Commun. 3, 986 (2012).

    PubMed  Google Scholar 

  21. Graham, J.B., Da Costa, A. & Lund, J.M. Regulatory T cells shape the resident memory T cell response to virus infection in the tissues. J. Immunol. 192, 683–690 (2014).

    CAS  PubMed  Google Scholar 

  22. Kühn, R., Löhler, J., Rennick, D., Rajewsky, K. & Müller, W. Interleukin-10-deficient mice develop chronic enterocolitis. Cell 75, 263–274 (1993).

    PubMed  Google Scholar 

  23. Maharshak, N. et al. Altered enteric microbiota ecology in interleukin 10-deficient mice during development and progression of intestinal inflammation. Gut Microbes 4, 316–324 (2013).

    PubMed  PubMed Central  Google Scholar 

  24. Stelekati, E. & Wherry, E.J. Chronic bystander infections and immunity to unrelated antigens. Cell Host Microbe 12, 458–469 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Ouyang, W., Rutz, S., Crellin, N.K., Valdez, P.A. & Hymowitz, S.G. Regulation and functions of the IL-10 family of cytokines in inflammation and disease. Annu. Rev. Immunol. 29, 71–109 (2011).

    CAS  PubMed  Google Scholar 

  26. Cui, W., Joshi, N.S., Jiang, A. & Kaech, S.M. Effects of Signal 3 during CD8 T cell priming: Bystander production of IL-12 enhances effector T cell expansion but promotes terminal differentiation. Vaccine 27, 2177–2187 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Parish, I.A. et al. Chronic viral infection promotes sustained Th1-derived immunoregulatory IL-10 via BLIMP-1. J. Clin. Invest. 124, 3455–3468 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Brooks, D.G. et al. Interleukin-10 determines viral clearance or persistence in vivo. Nat. Med. 12, 1301–1309 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Maynard, C.L. et al. Regulatory T cells expressing interleukin 10 develop from Foxp3+ and Foxp3− precursor cells in the absence of interleukin 10. Nat. Immunol. 8, 931–941 (2007).

    CAS  PubMed  Google Scholar 

  30. Srivastava, S., Koch, M.A., Pepper, M. & Campbell, D.J. Type I interferons directly inhibit regulatory T cells to allow optimal antiviral T cell responses during acute LCMV infection. J. Exp. Med. 211, 961–974 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Cheng, G. et al. IL-2 receptor signaling is essential for the development of Klrg1+ terminally differentiated T regulatory cells. J. Immunol. 189, 1780–1791 (2012).

    CAS  PubMed  Google Scholar 

  32. Steinman, R.M., Pack, M. & Inaba, K. Dendritic cells in the T-cell areas of lymphoid organs. Immunol. Rev. 156, 25–37 (1997).

    CAS  PubMed  Google Scholar 

  33. Jung, Y.W., Rutishauser, R.L., Joshi, N.S., Haberman, A.M. & Kaech, S.M. Differential localization of effector and memory CD8 T cell subsets in lymphoid organs during acute viral infection. J. Immunol. 185, 5315–5325 (2010).

    CAS  PubMed  Google Scholar 

  34. Rubtsov, Y.P. et al. Regulatory T cell-derived interleukin-10 limits inflammation at environmental interfaces. Immunity 28, 546–558 (2008).

    CAS  PubMed  Google Scholar 

  35. Kaech, S.M., Hemby, S., Kersh, E. & Ahmed, R. Molecular and functional profiling of memory CD8 T cell differentiation. Cell 111, 837–851 (2002).

    CAS  PubMed  Google Scholar 

  36. Luckey, C.J. et al. Memory T and memory B cells share a transcriptional program of self-renewal with long-term hematopoietic stem cells. Proc. Natl. Acad. Sci. USA 103, 3304–3309 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Agarwal, P. et al. Gene regulation and chromatin remodeling by IL-12 and type I IFN in programming for CD8 T cell effector function and memory. J. Immunol. 183, 1695–1704 (2009).

    CAS  PubMed  Google Scholar 

  38. Amit, I. et al. Unbiased reconstruction of a mammalian transcriptional network mediating pathogen responses. Science 326, 257–263 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Girard-Madoux, M.J.H., Kel, J.M., Reizis, B. & Clausen, B.E. IL-10 controls dendritic cell-induced T-cell reactivation in the skin to limit contact hypersensitivity. J. Allergy Clin. Immunol. 129, 143–150.e1–10 (2012).

    CAS  PubMed  Google Scholar 

  40. Kim, J.M., Rasmussen, J.P. & Rudensky, A.Y. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice. Nat. Immunol. 8, 191–197 (2007).

    CAS  PubMed  Google Scholar 

  41. Onishi, Y., Fehervari, Z., Yamaguchi, T. & Sakaguchi, S. Foxp3+ natural regulatory T cells preferentially form aggregates on dendritic cells in vitro and actively inhibit their maturation. Proc. Natl. Acad. Sci. USA 105, 10113–10118 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Borsellino, G. et al. Expression of ectonucleotidase CD39 by Foxp3+ Treg cells: hydrolysis of extracellular ATP and immune suppression. Blood 110, 1225–1232 (2007).

    CAS  PubMed  Google Scholar 

  43. Fallarino, F. et al. Modulation of tryptophan catabolism by regulatory T cells. Nat. Immunol. 4, 1206–1212 (2003).

    CAS  PubMed  Google Scholar 

  44. Hochrein, H. et al. Differential production of IL-12, IFN-α, and IFN-γ by mouse dendritic cell subsets. J. Immunol. 166, 5448–5455 (2001).

    CAS  PubMed  Google Scholar 

  45. Moser, E.K., Hufford, M.M. & Braciale, T.J. Late engagement of CD86 after influenza virus clearance promotes recovery in a FoxP3+ regulatory T cell dependent manner. PLoS Pathog. 10, e1004315 (2014).

    PubMed  PubMed Central  Google Scholar 

  46. Liston, A. & Gray, D.H.D. Homeostatic control of regulatory T cell diversity. Nat. Rev. Immunol. 14, 154–165 (2014).

    CAS  PubMed  Google Scholar 

  47. Zeiser, R. et al. Differential impact of mammalian target of rapamycin inhibition on CD4+CD25+Foxp3+ regulatory T cells compared with conventional CD4+ T cells. Blood 111, 453–462 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  48. Keating, R. et al. The kinase mTOR modulates the antibody response to provide cross-protective immunity to lethal infection with influenza virus. Nat. Immunol. 14, 1266–1276 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Cui, W. et al. TLR4 ligands lipopolysaccharide and monophosphoryl lipid a differentially regulate effector and memory CD8+ T Cell differentiation. J. Immunol. 192, 4221–4232 (2014).

    CAS  PubMed  Google Scholar 

  50. Best, J.A. et al. Transcriptional insights into the CD8+ T cell response to infection and memory T cell formation. Nat. Immunol. 14, 404–412 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  51. Kühn, R., Löhler, J., Rennick, D., Rajewsky, K. & Müller, W. Interleukin-10-deficient mice develop chronic enterocolitis. Cell 75, 263–274 (1993).

    PubMed  Google Scholar 

  52. Maynard, C.L. et al. Regulatory T cells expressing interleukin 10 develop from Foxp3+ and Foxp3− precursor cells in the absence of interleukin 10. Nat. Immunol. 8, 931–941 (2007).

    CAS  PubMed  Google Scholar 

  53. Roers, A. et al. T cell-specific inactivation of the interleukin 10 gene in mice results in enhanced T cell responses but normal innate responses to lipopolysaccharide or skin irritation. J. Exp. Med. 200, 1289–1297 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  54. Kaech, S.M. et al. Selective expression of the interleukin 7 receptor identifies effector CD8 T cells that give rise to long-lived memory cells. Nat. Immunol. 4, 1191–1198 (2003).

    CAS  PubMed  Google Scholar 

  55. Kim, J.M., Rasmussen, J.P. & Rudensky, A.Y. Regulatory T cells prevent catastrophic autoimmunity throughout the lifespan of mice. Nat. Immunol. 8, 191–197 (2007).

    CAS  PubMed  Google Scholar 

  56. Joshi, N.S. et al. Inflammation directs memory precursor and short-lived effector CD8+ T cell fates via the graded expression of T-bet transcription factor. Immunity 27, 281–295 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  57. Teijaro, J.R. et al. Cutting edge: Tissue-retentive lung memory CD4 T cells mediate optimal protection to respiratory virus infection. J. Immunol. 187, 5510–5514 (2011).

    CAS  PubMed  Google Scholar 

  58. Anders, S. et al. Count-based differential expression analysis of RNA sequencing data using R and Bioconductor. Nat. Protoc. 8, 1765–1786 (2013).

    PubMed  Google Scholar 

  59. Love, M.I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 550 (2014).

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank J.M.M. den Haan (UV University Medical Center, Amsterdam) for mAb ES5-2A5; and all members of the Kaech and Craft laboratories for discussions and critical reading of the manuscript. Supported by the US National Institutes of Health (RO1AI066232 and R01AI074699 to S.M.K.; R01AR40072, P30AR053495 and R21AR063942 to J.C.; T32AI07019 and F31AG07777 to B.J.L.; and T32GM07205 to S.M.G.) and the Howard Hughes Medical Institute (S.M.K., B.J.L., R.A.F. and T.G.).

Author information

Author notes

  1. Weiguo Cui

    Present address: Present address: Blood Research Institute, BloodCenter of Wisconsin, Milwaukee, Wisconsin, USA.,

  2. Joe Craft and Susan M Kaech: These authors jointly directed this work.

Authors and Affiliations

  1. Department of Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA

    Brian J Laidlaw, Weiguo Cui, Robert A Amezquita, Simon M Gray, Tianxia Guan, Yisi Lu, Yasushi Kobayashi, Richard A Flavell, Steven H Kleinstein, Joe Craft & Susan M Kaech

  2. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA

    Robert A Amezquita, Richard A Flavell & Susan M Kaech

  3. Department of Pathology, Yale School of Medicine, New Haven, Connecticut, USA

    Steven H Kleinstein

  4. Interdepartmental Program in Computational Biology and Bioinformatics, Yale School of Medicine, New Haven, Connecticut, USA

    Steven H Kleinstein

  5. Department of Internal Medicine (Rheumatology), Yale School of Medicine, New Haven, Connecticut, USA

    Joe Craft

Authors
  1. Brian J Laidlaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weiguo Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robert A Amezquita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Simon M Gray
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tianxia Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yisi Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yasushi Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Richard A Flavell
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Steven H Kleinstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Joe Craft
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Susan M Kaech
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

B.J.L., J.C., S.M.K. conceived of and designed the experiments, analyzed the data and wrote the manuscript; and B.J.L., W.C., R.A.A., S.M.G., T.G., Y.L., Y.K., S.H.K. and R.A.F. performed the experiments.

Corresponding authors

Correspondence to Joe Craft or Susan M Kaech.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 IL-10 is required for optimal maturation of memory CD8+ NP396+ T cells.

(a) Analysis of the virus-specific CD8+ T cell response 60 days post acute LCMV Armstrong infection. The percentages and numbers of GP33+ T cells along with the (b) representative plots and analysis of the NP396+ T cell response in the spleen of wild type and Il10–/– mice are shown. Percentages and numbers of NP396+ T cells, along with the percentages of CD127+KLRG1-, CD127-KLRG1+, and CD62L+KLRG1- cells in the NP396+ T cell population are shown. (c) Mice were infected with acute LCMV and treated with αIL-10 between days 0-30, 0-8, 8-30, 15-30 or mock injected with PBS. Mice were sacrificed at day 30 and the percentage and numbers of GP33+ T cells determined. Statistical analyses were performed using the unpaired two-tailed Student’s t-test. (*, p < 0.01; **, p < 0.001). Data are from one experiment representative of 5 experiments (a, b) or 3 experiments (c) with at least 4 mice per group carried out 45-60 days (a, b) or 30 days (c) following LCMV Armstrong infection (mean and s.e.m).

Supplementary Figure 2 IL-10 is required in a CD8+ T cell–extrinsic and CD8+ T cell–intrinsic manner to allow the maturation and survival of memory CD8+ T cells.

50,000 Il10raf/f or Il10raf/fCd4-Cre P14 GP33+ cells were transferred into congenically-mismatched mice one day prior to infection with acute LCMV infection. Analysis of the P14 GP33+ T cell response was carried out 30 days p.i. (a) Representative plots and analysis of the P14 GP33+ T cell response. Percentages and numbers of P14 GP33+ T cells, along with the percentages of CD127+KLRG1-, CD127-KLRG1+, and CD62L+KLRG1- cells in the P14 GP33+ T cell population are shown. (b) Representative histograms of GzmB and Tcf1 expression in Il10raf/f (black) or Il10raf/fCd4-Cre (gray filled) P14 GP33+ cells. Statistical analyses were performed using the unpaired two-tailed Student’s t-test. (*, p < 0.05; **, p < 0.01). Data are from one experiment representative of 2 experiments with 4 mice per group carried out 30 days following LCMV Armstrong infection (mean and s.e.m).

Supplementary Figure 3 CD4+ Treg cells continue to produce IL-10 during the resolution phase of infection.

(a) Analysis of the GP33+ T cell response 60 days post acute LCMV infection in Il10f/f, Il10f/fCd4-Cre, Il10f/fLyz2-Cre, and Il10f/fCd11c-Cre mice. Percentages and numbers of GP33+ T cells were determined. (b) Analysis of IL-10 production by T cells following acute LCMV infection in IL-10 reporter (10BiT Thy1.1 mice). Percentages and numbers of IL-10-Thy1.1 reporter-positive cells at multiple time points post LCMV infection are shown. (c) Intravenously (i.v.) administered anti-CD4 antibody was used to distinguish circulating (red pulp localized) versus resident (white pulp localized) CD4+ T cells. (d) Analysis of the GP33+ T cell response 60 days post acute LCMV Armstrong infection. Percentages and numbers of GP33+ T cells in Il10f/f and Il10f/f Foxp3-Cre mice are shown. Data are from one experiment representative of 3 experiments (a, b, c, d) with 3-6 mice per group carried out 45-60 days (a, d), or 0, 8, and 15 days (b, c) following LCMV Armstrong infection (mean and s.e.m).

Supplementary Figure 4 Validation of RNA-seq results.

(a) Expression of Zeb2, Ccr7, Cx3cr1, and Pim1 was determined by qPCR analysis of cDNA isolated from pooled GP33+ and NP396+ CD8+ T cells at 15 days post acute LCMV infection from Il10f/f and Il10f/f Foxp3-Cre mice. Expression of TCF-1 and GzmB in GP33+ in T cells at 15 days post acute LCMV infection from Il10f/f and Il10f/f Foxp3-Cre mice was determined by flow cytometric analysis. (b) Representative plots of the GP33+ T cell response in Il10f/f and Il10f/f Foxp3Cre mice at day 15 post LCMV infection. Data are from one experiment representative of 3 experiments with 3-5 mice per group carried out 15 days following LCMV Armstrong infection (mean and s.e.m).

Supplementary Figure 5 Heat map of differentially expressed genes based on RNA-seq results.

Genes with a p-adjusted value < 0.2 (Benjami-Hochberg) and the corresponding log2 fold-change in mRNA isolated from pooled GP33+ and NP396+ CD8+ T cells at 15 days post acute LCMV infection from Il10f/f and Il10f/f Foxp3-Cre mice.

Supplementary Figure 6 Virus-specific CD8+ T cells from mice lacking CD4+ Treg cell–derived IL-10 display a robust inflammatory and effector gene signature.

mRNA was isolated from pooled GP33+ and NP396+ CD8+ T cells at 15 days post acute LCMV infection from Il10f/f and Il10f/f Foxp3-Cre mice and compared by RNA-seq. (a) Gene set plots showing individual log2 fold-changes of with corresponding standard error based on published effector vs memory gene set. Gene Set Enrichment Analysis (GSEA) was performed using gene sets from the Broad MSigDB collection; select significantly enriched gene sets (FDR < 1e-5) are shown with their running Enrichment Score (ES) (line), where members of the gene set appear in the ranked list of genes (barcode), and the signal to noise ranking metric (bar). A positive ES signifies enrichment in the Il10f/f Foxp3-Cre sample relative to the WT condition of a given gene set; i.e., more highly expressed. (b) GSEA results of CpG (c) and poly:IC stimulated genes (bottom) were visualized. (d) Normalized enrichment scores for Gene Set Enrichment Analysis. Normalized enrichment scores (NES) was calculated for select significantly enriched gene sets (FDR < 1e-5). Gene set name, figure GSEA plots shown in, and NES are shown in table.

Supplementary Figure 7 Transfer of IL-10-competent CD4+ Treg cells during the resolution phase of LCMV infection is sufficient to ‘rescue’ the maturation defect of memory CD8+ T cells in Il10–/– mice.

Analysis of the GP33+ T cell response 60 days p.i. in Foxp3GFP-DTR mice treated with diphtheria toxin at day -1, day 8, or day 15 p.i. or mock injected with PBS. (a) Percentage and numbers of GP33+ T cells are shown. Representative of 3 independent experiments with 3-6 mice per group carried out 45-60 days following LCMV Armstrong infection. (b) Analysis of the GP33+ T cell response 60 days post acute LCMV Armstrong infection in Il10–/– mice and Il10–/– mice that were administered 3x105 Foxp3+ CD4+ T cells isolated from coinfected Foxp3GFP-DTR mice at day 8 p.i. Percentage and numbers of GP33+ T cells are shown. Representative of 2 independent experiments with 3-7 mice per group carried out 60 days following LCMV Armstrong infection. Data are from one experiment representative of 3 experiments (a) or 2 experiments (b) with 3-7 mice per group carried out 45-60 days following LCMV Armstrong infection (mean and s.e.m).

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 (PDF 993 kb)

Source data

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Laidlaw, B., Cui, W., Amezquita, R. et al. Production of IL-10 by CD4+ regulatory T cells during the resolution of infection promotes the maturation of memory CD8+ T cells. Nat Immunol 16, 871–879 (2015). https://doi.org/10.1038/ni.3224

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.3224

This article is cited by

Search

Advanced 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