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Proinflammatory microenvironments within the intestine regulate the differentiation of tissue-resident CD8+ T cells responding to infection

Abstract

We report that oral infection with Yersinia pseudotuberculosis results in the development of two distinct populations of pathogen-specific CD8+ tissue-resident memory T cells (TRM cells) in the lamina propria. CD103 T cells did not require transforming growth factor-β (TGF-β) signaling but were true resident memory cells. Unlike CD103+CD8+ T cells, which were TGF-β dependent and were scattered in the tissue, CD103CD8+ T cells clustered with CD4+ T cells and CX3CR1+ macrophages and/or dendritic cells around areas of bacterial infection. CXCR3-dependent recruitment of cells to inflamed areas was critical for development of the CD103 population and pathogen clearance. Our studies have identified the 'preferential' development of CD103 TRM cells in inflammatory microenvironments within the lamina propria and suggest that this subset has a critical role in controlling infection.

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Figure 1: Oral Yptb-OVA infection generates a robust intestinal CD8+ T cell response.
Figure 2: CD103CD8+ populations develop in the lamina propria and have features of resident memory cells.
Figure 3: TGF-β signaling is not required for development of the lamina propria CD103CD8+ TRM cell population.
Figure 4: Yptb-OVA infection stimulates the formation of CD103CD8+ T cell clusters in the lamina propria.
Figure 5: Yptb-OVA–induced clusters contain T cells and CX3CR1+ macrophages and/or DCs, but not B cells.
Figure 6: CX3CR1-expressing macrophage and/or DC populations present Yptb-OVA antigens, but antigenic stimulation does not downregulate CD103 expression on intestinal TRM cells.
Figure 7: Cxcr3-deficient T cells enter the intestine but fail to localize to areas of inflammation.
Figure 8: CXCR3-mediated localization affects CD103 expression on intestinal CD8+ TRM cells and protection.

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References

  1. Masopust, D., Vezys, V., Marzo, A.L. & Lefrancois, L. Preferential localization of effector memory cells in nonlymphoid tissue. Science 291, 2413–2417 (2001).

    CAS  PubMed  Google Scholar 

  2. Johansson-Lindbom, B. et al. Selective generation of gut tropic T cells in gut-associated lymphoid tissue (GALT): requirement for GALT dendritic cells and adjuvant. J. Exp. Med. 198, 963–969 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Mora, J.R. et al. Selective imprinting of gut-homing T cells by Peyer's patch dendritic cells. Nature 424, 88–93 (2003).

    CAS  PubMed  Google Scholar 

  4. Masopust, D. et al. Dynamic T cell migration program provides resident memory within intestinal epithelium. J. Exp. Med. 207, 553–564 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Masopust, D., Vezys, V., Wherry, E.J., Barber, D.L. & Ahmed, R. Cutting edge: gut microenvironment promotes differentiation of a unique memory CD8 T cell population. J. Immunol. 176, 2079–2083 (2006).

    CAS  PubMed  Google Scholar 

  6. Mueller, S.N., Gebhardt, T., Carbone, F.R. & Heath, W.R. Memory T cell subsets, migration patterns, and tissue residence. Annu. Rev. Immunol. 31, 137–161 (2013).

    CAS  PubMed  Google Scholar 

  7. Sheridan, B.S. et al. Oral infection drives a distinct population of intestinal resident memory CD8(+) T cells with enhanced protective function. Immunity 40, 747–757 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Mackay, L.K. et al. The developmental pathway for CD103(+)CD8(+) tissue-resident memory T cells of skin. Nat. Immunol. 14, 1294–1301 (2013).

    CAS  PubMed  Google Scholar 

  9. Wakim, L.M. et al. The molecular signature of tissue resident memory CD8 T cells isolated from the brain. J. Immunol. 189, 3462–3471 (2012).

    CAS  PubMed  Google Scholar 

  10. Casey, K.A. et al. Antigen-independent differentiation and maintenance of effector-like resident memory T cells in tissues. J. Immunol. 188, 4866–4875 (2012).

    CAS  PubMed  Google Scholar 

  11. Zhang, N. & Bevan, M.J. Transforming growth factor-β signaling controls the formation and maintenance of gut-resident memory T cells by regulating migration and retention. Immunity 39, 687–696 (2013).

    PubMed  PubMed Central  Google Scholar 

  12. Skon, C.N. et al. Transcriptional downregulation of S1pr1 is required for the establishment of resident memory CD8+ T cells. Nat. Immunol. 14, 1285–1293 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Bergman, M.A., Loomis, W.P., Mecsas, J., Starnbach, M.N. & Isberg, R.R. CD8(+) T cells restrict Yersinia pseudotuberculosis infection: bypass of anti-phagocytosis by targeting antigen-presenting cells. PLoS Pathog. 5, e1000573 (2009).

    PubMed  PubMed Central  Google Scholar 

  14. Lin, J.-S., Szaba, F.M., Kummer, L.W., Chromy, B.A. & Smiley, S.T. Yersinia pestis YopE contains a dominant CD8 T cell epitope that confers protection in a mouse model of pneumonic plague. J. Immunol. 187, 897–904 (2011).

    CAS  PubMed  Google Scholar 

  15. Zhang, Y. et al. A protective epitope in type iii effector YopE is a major CD8 T cell antigen during primary infection with Yersinia pseudotuberculosis. Infect. Immun. 80, 206–214 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Barnes, P.D., Bergman, M.A., Mecsas, J. & Isberg, R.R. Yersinia pseudotuberculosis disseminates directly from a replicating bacterial pool in the intestine. J. Exp. Med. 203, 1591–1601 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Melton-Witt, J.A., Rafelski, S.M., Portnoy, D.A. & Bakardjiev, A.I. Oral infection with signature-tagged listeria monocytogenes reveals organ-specific growth and dissemination routes in guinea pigs. Infect. Immun. 80, 720–732 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Wiedig, C.A., Kramer, U., Garbom, S., Wolf-Watz, H. & Autenrieth, I.B. Induction of CD8(+) T cell responses by Yersinia vaccine carrier strains. Vaccine 23, 4984–4998 (2005).

    CAS  PubMed  Google Scholar 

  19. Logsdon, L.K. & Mecsas, J. Requirement of the Yersinia pseudotuberculosis effectors YopH and YopE in colonization and persistence in intestinal and lymph tissues. Infect. Immun. 71, 4595–4607 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Cornelis, G.R. et al. The virulence plasmid of Yersinia, an antihost genome. Microbiol. Mol. Biol. Rev. 62, 1315–1352 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Wakim, L.M., Woodward-Davis, A. & Bevan, M.J. Memory T cells persisting within the brain after local infection show functional adaptations to their tissue of residence. Proc. Natl. Acad. Sci. USA 107, 17872–17879 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. El-Asady, R. et al. TGF-beta-dependent CD103 expression by CD8(+) T cells promotes selective destruction of the host intestinal epithelium during graft-versus-host disease. J. Exp. Med. 201, 1647–1657 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Bouskra, D. et al. Lymphoid tissue genesis induced by commensals through NOD1 regulates intestinal homeostasis. Nature 456, 507–510 (2008).

    CAS  PubMed  Google Scholar 

  24. Clark, M.A., Hirst, B.H. & Jepson, M.A. M-cell surface beta 1 integrin expression and invasin-mediated targeting of Yersinia pseudotuberculosis to mouse Peyer's patch M cells. Infect. Immun. 66, 1237–1243 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Jang, M.H. et al. Intestinal villous M cells: an antigen entry site in the mucosal epithelium. Proc. Natl. Acad. Sci. USA 101, 6110–6115 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Halle, S. et al. Solitary intestinal lymphoid tissue provides a productive port of entry for Salmonella enterica serovar Typhimurium. Infect. Immun. 75, 1577–1585 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Autenrieth, I.B., Tingle, A., Reske-Kunz, A. & Heesemann, J. T lymphocytes mediate protection against Yersinia enterocolitica in mice: characterization of murine T-cell clones specific for Y. enterocolitica. Infect. Immun. 60, 1140–1149 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Philipovskiy, A.V. & Smiley, S.T. Vaccination with live Yersinia pestis primes CD4 and CD8 T cells that synergistically protect against lethal pulmonary Y. pestis infection. Infect. Immun. 75, 878–885 (2007).

    CAS  PubMed  Google Scholar 

  29. Parent, M.A. et al. Cell-mediated protection against pulmonary Yersinia pestis infection. Infect. Immun. 73, 7304–7310 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Zigmond, E. et al. Ly6C hi monocytes in the inflamed colon give rise to proinflammatory effector cells and migratory antigen-presenting cells. Immunity 37, 1076–1090 (2012).

    CAS  PubMed  Google Scholar 

  31. Mackay, L.K. et al. Long-lived epithelial immunity by tissue-resident memory T (TRM) cells in the absence of persisting local antigen presentation. Proc. Natl. Acad. Sci. USA 109, 7037–7042 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Schulz, O. et al. Intestinal CD103+, but not CX3CR1+, antigen sampling cells migrate in lymph and serve classical dendritic cell functions. J. Exp. Med. 206, 3101–3114 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Kohlmeier, J.E. et al. Inflammatory chemokine receptors regulate CD8(+) T cell contraction and memory generation following infection. J. Exp. Med. 208, 1621–1634 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Harris, T.H. et al. Generalized Lévy walks and the role of chemokines in migration of effector CD8+ T cells. Nature 486, 545–548 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Nakanishi, Y., Lu, B., Gerard, C. & Iwasaki, A. CD8(+) T lymphocyte mobilization to virus-infected tissue requires CD4(+) T-cell help. Nature 462, 510–513 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Schenkel, J.M., Fraser, K.A., Vezys, V. & Masopust, D. Sensing and alarm function of resident memory CD8(+) T cells. Nat. Immunol. 14, 509–513 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Groom, J.R. & Luster, A.D. CXCR3 ligands: redundant, collaborative and antagonistic functions. Immunol. Cell Biol. 89, 207–215 (2011).

    CAS  PubMed  Google Scholar 

  38. Kurachi, M. et al. Chemokine receptor CXCR3 facilitates CD8(+) T cell differentiation into short-lived effector cells leading to memory degeneration. J. Exp. Med. 208, 1605–1620 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Hu, J.K., Kagari, T. & Clingan, J.M. Expression of chemokine receptor CXCR3 on T cells affects the balance between effector and memory CD8 T-cell generation. Proc. Natl. Acad. Sci. USA 108, E118–E127 (2011).

    PubMed  PubMed Central  Google Scholar 

  40. Travis, M.A. & Sheppard, D. TGF-β activation and function in immunity. Annu. Rev. Immunol. 32, 51–82 (2014).

    CAS  PubMed  Google Scholar 

  41. Dube, P.H., Revell, P.A., Chaplin, D.D., Lorenz, R.G. & Miller, V.L. A role for IL-1 alpha in inducing pathologic inflammation during bacterial infection. Proc. Natl. Acad. Sci. USA 98, 10880–10885 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  42. DePaolo, R.W. et al. A specific role for TLR1 in protective T(H)17 immunity during mucosal infection. J. Exp. Med. 209, 1437–1444 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Dube, P.H., Handley, S.A., Lewis, J. & Miller, V.L. Protective role of interleukin-6 during Yersinia enterocolitica infection is mediated through the modulation of inflammatory cytokines. Infect. Immun. 72, 3561–3570 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Jung, C. et al. Yersinia pseudotuberculosis disrupts intestinal barrier integrity through hematopoietic TLR-2 signaling. J. Clin. Invest. 122, 2239–2251 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Schenkel, J.M. et al. T cell memory. Resident memory CD8 T cells trigger protective innate and adaptive immune responses. Science 346, 98–101 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Ariotti, S. et al. T cell memory. Skin-resident memory CD8+ T cells trigger a state of tissue-wide pathogen alert. Science 346, 101–105 (2014).

    CAS  PubMed  Google Scholar 

  47. Gebhardt, T. et al. Different patterns of peripheral migration by memory CD4(+) and CD8(+) T cells. Nature 477, 216–219 (2011).

    CAS  PubMed  Google Scholar 

  48. Bergsbaken, T. & Cookson, B.T. Macrophage activation redirects Yersinia-infected host cell death from apoptosis to caspase-1-dependent pyroptosis. PLoS Pathog. 3, e161 (2007).

    PubMed  PubMed Central  Google Scholar 

  49. Viboud, G.I., So, S., Ryndak, M.B. & Bliska, J.B. Proinflammatory signalling stimulated by the type III translocation factor YopB is counteracted by multiple effectors in epithelial cells infected with Yersinia pseudotuberculosis. Mol. Microbiol. 47, 1305–1315 (2003).

    CAS  PubMed  Google Scholar 

  50. Palmer, L.E., Hobbie, S., Galan, J.E. & Bliska, J.B. YopJ of Yersinia pseudotuberculosis is required for the inhibition of macrophage TNF-alpha production and downregulation of the MAP kinases p38 and JNK. Mol. Microbiol. 27, 953–965 (1998).

    CAS  PubMed  Google Scholar 

  51. Zhang, N. & Bevan, M.J. TGF-β signaling to T cells inhibits autoimmunity during lymphopenia-driven proliferation. Nat. Immunol. 13, 667–673 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  52. Moolenbeek, C. & Ruitenberg, E.J. The 'Swiss roll': a simple technique for histological studies of the rodent intestine. Lab. Anim. 15, 57–59 (1981).

    CAS  PubMed  Google Scholar 

  53. Sung, J.H. et al. Chemokine guidance of central memory T cells is critical for antiviral recall responses in lymph nodes. Cell 150, 1249–1263 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank M. Chase and X. Cun-Pan for technical assistance and N. Zhang for critical reading of the manuscript. Supported by the Howard Hughes Medical Institute, the US National Institutes of Health (AI-19335 to M.J.B.) and the Cancer Research Institute (T.B.).

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T.B. and M.J.B. designed experiments and wrote the manuscript; T.B. performed experiments and analyzed data.

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Correspondence to Michael J Bevan.

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Integrated supplementary information

Supplementary Figure 1 Generation of Yptb-OVA and verification of protein production and OT-I priming.

(a) Organization of manipulated genes on pIB1 virulence plasmid. (b) Western blot of supernatants from Yptb-OVA and Yptb-NEG strains using antibodies to ovalbumin. (c) YopE-specific and OT-I responses in the spleen on day 10 after infection with Yptb-OVA or Yptb-NEG. Plots are of CD8β+ gate. (d) Mice received OT-I T cells and were immunized with VSV-OVA. Sixty days after VSV-OVA infection, mice were infected with 2 × 108 Yptb-OVA or Yptb-NEG, and body weight was monitored. All mice survived Yptb-OVA infection, whereas 40% of mice infected with Yptb-NEG lost more than 20% of their body weight and had to be euthanized.

Supplementary Figure 2 The ratio of YopE-tet to OT-I T cells in the lamina propria remains stable over time.

Mice received OT-I T cells followed by Yptb-OVA. The ratio of YopE-species CD8+ T cells to OT-I T cells was analyzed at the indicated time points after infection. Data are mean and s.d. with three mice per group.

Supplementary Figure 3 S1pr1 expression is elevated in CD103 lamina propria populations.

The indicated CD8+ populations were sorted from Yptb-OVA–infected mice at more than 28 d after infection. RNA was isolated, and S1pr1 expression was determined using qRT-PCR; levels were normalized to Actb expression. *P < 0.05, **P < 0.005 compared to splenic CD8+ cells.

Supplementary Figure 4 OT-I clustering and Yptb-OVA colonization in different regions of the intestine.

(ad) C57BL/6 mice received 1 × 104 GFP+ OT-I T cells and were infected with Yptb-OVA. The intestine was isolated at 9 d after infection, and tissue sections were analyzed by immunohistochemistry. (a) The percentage of CD103+ OT-I cells in individual clusters was enumerated by microscopy and compared to the total in the lamina propria as measured by FACS. *P < 0.0001. Representative images from three mice showing the distribution of OT-I cells in the cecum (b), colon (c), and duodenum (d) of the small intestine. (e) C57BL/6 mice were infected with Yptb-OVA, and at 6 d after infection the small intestine, colon, and cecum were isolated. The small intestine was divided into three equal parts, and tissues were washed, homogenized, and plated on CIN agar. Data are expressed as the total number of colony-forming units (CFU). Data pooled from multiple experiments; dark symbols indicate that the number of CFU was below the limit of detection.

Supplementary Figure 5 CD8+ clusters do not contain B220+ cells.

C57BL/6 mice were infected with Yptb-OVA, and at 9 d after infection the small intestine was isolated and tissue sections were analyzed by immunohistochemistry. Representative image from three mice.

Supplementary Figure 6 CX3CR1-GFPint cells have a phenotype consistent with recently recruited inflammatory monocytes.

Cx3cr1gfp/+ mice were infected with Yptb-OVA, and on day 7 after infection lamina propria cells were isolated and the expression of CCR2, Ly6C, and F4/80 was examined on CX3CR1hi and CX3CR1int APC populations.

Supplementary Figure 7 Clusters persist after infection is cleared but diminish over time.

(a) C57BL/6 mice were infected with Yptb-OVA, and at 6 d after infection the ileum was isolated. Tissues were washed, homogenized, and plated on CIN agar. Data are expressed as the total number of colony-forming units (CFU). Data pooled from multiple experiments. Dark symbols indicate that the number of CFU was below the limit of detection. (bd) C57BL/6 mice received 1 × 104 GFP+ OT-I T cells and were infected with Yptb-OVA. The intestine was isolated at 30 or 120 d after infection, tissue sections were analyzed by immunohistochemistry (c,d), and the number of OT-I cells per villus was determined (b). Representative images from three mice.

Supplementary Figure 8 Altered peripheral activation/stimulation of Cxcr3-deficient OT-I cells in vivo does not affect CD103 expression.

(a) Mice received 5 × 103 each of wild-type and Cxcr3-deficient OT-I cells. On day 7 after infection, the percentage of OT-I cells with SLEC (KLRG1+CD127) and MPEC (KLRG1CD127+) phenotypes in the MLN was determined. *P < 0.05, **P < 0.005. (b) Day 5 Yptb-OVA mice received 1 × 106 each of wild-type and Cxcr3-deficient in vitro activated OT-I T cells. Four days after cell transfer, the expression of CD103 was examined on lamina propria OT-I cells. Lines connect cell populations from the same mouse, pooled from three experiments. *P < 0.05. (c) Cells were isolated from the MLN on day 5 after infection and stimulated in vitro with TGFβ and CXCL10 for 20 h or were left untreated, and CD103 expression on OT-I T cells was assessed. Representative of three experiments.

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Bergsbaken, T., Bevan, M. Proinflammatory microenvironments within the intestine regulate the differentiation of tissue-resident CD8+ T cells responding to infection. Nat Immunol 16, 406–414 (2015). https://doi.org/10.1038/ni.3108

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