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.

Peripherally induced brain tissue–resident memory CD8+ T cells mediate protection against CNS infection

Abstract

The central nervous system (CNS) is classically viewed as immune-privileged; however, recent advances highlight interactions between the peripheral immune system and CNS in controlling infections and tissue homeostasis. Tissue-resident memory (TRM) CD8+ T cells in the CNS are generated after brain infections, but it is unknown whether CNS infection is required to generate brain TRM cells. We show that peripheral infections generate antigen-specific CD8+ memory T cells in the brain that adopt a unique TRM signature. Upon depletion of circulating and perivascular memory T cells, this brain signature was enriched and the surveilling properties of brain TRM cells was revealed by intravital imaging. Notably, peripherally induced brain TRM cells showed evidence of rapid activation and enhanced cytokine production and mediated protection after brain infections. These data reveal that peripheral immunizations can generate brain TRM cells and will guide potential use of T cells as therapeutic strategies against CNS infections and neurological diseases.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: Enrichment of antigen-specific CD8+ T cells in the CNS after peripheral immunizations.
Fig. 2: Antigen-specific T cells in CNS adopt a tissue-resident memory phenotype after peripheral immunization.
Fig. 3: Increased representation of antigen-specific CD8+ T cells after peripheral immunizations is CNS specific.
Fig. 4: CD8+ T cell dynamics in CNS after peripheral immunization.
Fig. 5: Peripherally induced CNS TRM cells are resistant to systemic depletion.
Fig. 6: Enhanced effector functions of peripherally induced TRM CD8+ T cells in the CNS.
Fig. 7: Peripherally induced brain-resident CD8+ T cells mediate protection against CNS infections.

Data availability

The raw flow cytometric data that support the findings in SPADE analyses in Figs. 2, 3 and 5 are available from the corresponding author upon request. Source data for figures are provided with the paper. The RNA-seq data are deposited at the GEO with accession number GSE146077.

References

  1. 1.

    Louveau, A., Harris, T. H. & Kipnis, J. Revisiting the mechanisms of CNS immune privilege. Trends Immunol. 36, 569–577 (2015).

    PubMed  PubMed Central  CAS  Google Scholar 

  2. 2.

    Korn, T. & Kallies, A. T cell responses in the central nervous system. Nat. Rev. Immunol. 17, 179–194 (2017).

    PubMed  CAS  Google Scholar 

  3. 3.

    Manglani, M. & McGavern, D. B. New advances in CNS immunity against viral infection. Curr. Opin. Virol. 28, 116–126 (2018).

    PubMed  CAS  Google Scholar 

  4. 4.

    Aspelund, A. et al. A dural lymphatic vascular system that drains brain interstitial fluid and macromolecules. J. Exp. Med. 212, 991–999 (2015).

    PubMed  PubMed Central  CAS  Google Scholar 

  5. 5.

    Louveau, A. et al. Structural and functional features of central nervous system lymphatic vessels. Nature 523, 337–341 (2015).

    PubMed  PubMed Central  CAS  Google Scholar 

  6. 6.

    Kipnis, J. Multifaceted interactions between adaptive immunity and the central nervous system. Science 353, 766–771 (2016).

    PubMed  PubMed Central  CAS  Google Scholar 

  7. 7.

    Harty, J. T. & Badovinac, V. P. Shaping and reshaping CD8+ T cell memory. Nat. Rev. Immunol. 8, 107–119 (2008).

    PubMed  CAS  Google Scholar 

  8. 8.

    Martin, M. D. & Badovinac, V. P. Defining memory CD8+ T cell. Front. Immunol. 9, 2692 (2018).

    PubMed  PubMed Central  Google Scholar 

  9. 9.

    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).

    PubMed  PubMed Central  CAS  Google Scholar 

  10. 10.

    Slütter, B. et al. Dynamics of influenza-induced lung-resident memory T cells underlie waning heterosubtypic immunity. Sci. Immunol. 2, eaag2031 (2017).

    PubMed  PubMed Central  Google Scholar 

  11. 11.

    Steinbach, K., Vincenti, I. & Merkler, D. Resident-memory T cells in tissue-restricted immune responses: for better or worse? Front. Immunol. 9, 2827 (2018).

    PubMed  PubMed Central  Google Scholar 

  12. 12.

    Welten, S. P. M., Sandu, I., Baumann, N. S. & Oxenius, A. Memory CD8+ T cell inflation vs tissue‐resident memory T cells: same patrollers, same controllers? Immunol. Rev. 283, 161–175 (2018).

    PubMed  CAS  Google Scholar 

  13. 13.

    Griffin, D. E. & Metcalf, T. Clearance of virus infection from the CNS. Curr. Opin. Virol. 1, 216–221 (2011).

    PubMed  PubMed Central  Google Scholar 

  14. 14.

    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).

    PubMed  CAS  Google Scholar 

  15. 15.

    Landrith, T. A. et al. CD103+CD8+ T cells in the Toxoplasma-infected brain exhibit a tissue-resident memory transcriptional profile. Front. Immunol. 8, 335 (2017).

    PubMed  PubMed Central  Google Scholar 

  16. 16.

    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).

    PubMed  CAS  Google Scholar 

  17. 17.

    Steinbach, K. et al. Brain-resident memory T cells represent an autonomous cytotoxic barrier to viral infection. J. Exp. Med. 213, 1571–1587 (2016).

    PubMed  PubMed Central  CAS  Google Scholar 

  18. 18.

    Frost, E. L., Kersh, A. E., Evavold, B. D. & Lukacher, A. E. Cutting edge: resident memory CD8+ T cells express high-affinity TCRs. J. Immunol. 195, 3520–3524 (2015).

    PubMed  PubMed Central  CAS  Google Scholar 

  19. 19.

    Maru, S., Jin, G., Schell, T. D. & Lukacher, A. E. TCR stimulation strength is inversely associated with establishment of functional brain-resident memory CD8+ T cells during persistent viral infection. PLOS Pathog. 13, e1006318 (2017).

    PubMed  PubMed Central  Google Scholar 

  20. 20.

    Mockus, T. E. et al. CD+4 T cells control development and maintenance of brain-resident CD8+ T cells during polyomavirus infection. PLOS Pathog. 14, e1007365 (2018).

    PubMed  PubMed Central  Google Scholar 

  21. 21.

    Prasad, S., Hu, S., Sheng, W. S., Chauhan, P. & Lokensgard, J. R. Reactive glia promote development of CD103+CD69+CD8+T-cells through programmed cell death-ligand 1 (PD-L1). Immunity Inflamm. Dis. 6, 332–344 (2018).

    CAS  Google Scholar 

  22. 22.

    Prasad, S. et al. The PD-1: PD-L1 pathway promotes development of brain-resident memory T cells following acute viral encephalitis. J. Neuroinflam. 14, 82 (2017).

    Google Scholar 

  23. 23.

    Smolders, J. et al. Tissue-resident memory T cells populate the human brain. Nat. Comm. 9, 4593 (2018).

    Google Scholar 

  24. 24.

    Ritzel, R. M. et al. Age-associated resident memory CD8+ T cells in the central nervous system are primed to potentiate inflammation after ischemic brain injury. J. Immunol. 196, 3318–3330 (2016).

    PubMed  PubMed Central  CAS  Google Scholar 

  25. 25.

    Dulken, B. W. et al. Single-cell analysis reveals T cell infiltration in old neurogenic niches. Nature 571, 205–210 (2019).

    PubMed  PubMed Central  CAS  Google Scholar 

  26. 26.

    Gate, D. et al. Clonally expanded CD8+ T cells patrol the cerebrospinal fluid in Alzheimer’s disease. Nature 577, 399–404 (2020).

    PubMed  CAS  Google Scholar 

  27. 27.

    Prasad, S. & Lokensgard, J. R. Brain-resident T cells following viral infection. Viral Immunol. 32, 48–54 (2019).

    PubMed  PubMed Central  CAS  Google Scholar 

  28. 28.

    Wherry, E. J., Blattman, J. N., Murali-Krishna, K., van der Most, R. & Ahmed, R. Viral persistence alters CD8+ T cell immunodominance and tissue distribution and results in distinct stages of functional impairment. J. Virol. 77, 4911–4927 (2003).

    PubMed  PubMed Central  CAS  Google Scholar 

  29. 29.

    Mackay, L. K. et al. Hobit and Blimp1 instruct a universal transcriptional program of tissue residency in lymphocytes. Science 352, 459–463 (2016).

    PubMed  CAS  Google Scholar 

  30. 30.

    Milner, J. J. & Goldrath, A. W. Transcriptional programming of tissue-resident memory CD8+ T cells. Curr. Opin. Immunol. 51, 162–169 (2018).

    PubMed  PubMed Central  CAS  Google Scholar 

  31. 31.

    Topham, D. J. & Reilly, E. C. Tissue-resident memory CD8+ T cells: from phenotype to function. Front. Immunol. 9, 515 (2018).

    PubMed  PubMed Central  Google Scholar 

  32. 32.

    Milner, J. J. et al. Runx3 programs CD8+ T cell residency in non-lymphoid tissues and tumours. Nature 552, 253–257 (2017).

    PubMed  PubMed Central  CAS  Google Scholar 

  33. 33.

    Cheuk, S. et al. CD49a expression defines tissue-resident CD8+ T cells poised for cytotoxic function in human skin. Immunity 46, 287–300 (2017).

    PubMed  PubMed Central  CAS  Google Scholar 

  34. 34.

    Behr, F. M., Chuwonpad, A., Stark, R. & van Gisbergen, K. P. J. M. Armed and ready: transcriptional regulation of tissue-resident memory CD8+ T cells. Front. Immunol. 9, 1770 (2018).

    PubMed  PubMed Central  Google Scholar 

  35. 35.

    Mackay, L. K. & Kallies, A. Transcriptional regulation of tissue-resident lymphocytes. Trends Immunol. 38, 94–103 (2017).

    PubMed  CAS  Google Scholar 

  36. 36.

    Masopust, D. & Soerens, A. G. Tissue-resident T cells and other resident leukocytes. Ann. Rev. Immunol. 37, 521–546 (2019).

    CAS  Google Scholar 

  37. 37.

    McNamara, H. A. et al. Up-regulation of LFA-1 allows liver-resident memory T cells to patrol and remain in the hepatic sinusoids. Sci. Immunol. 2, eaaj1996 (2017).

    PubMed  PubMed Central  Google Scholar 

  38. 38.

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

    PubMed  CAS  Google Scholar 

  39. 39.

    Tse, S.-W., Radtke, A. J., Espinosa, D. A., Cockburn, I. A. & Zavala, F. The chemokine receptor CXCR6 is required for the maintenance of liver memory CD8+ T cells specific for infectious pathogens. J. Infect. Dis. 210, 1508–1516 (2014).

    PubMed  PubMed Central  CAS  Google Scholar 

  40. 40.

    Gerlach, C. et al. The chemokine receptor CX3CR1 defines three antigen-experienced CD8+ T cell subsets with distinct roles in immune surveillance and homeostasis. Immunity 45, 1270–1284 (2016).

    PubMed  PubMed Central  CAS  Google Scholar 

  41. 41.

    Obar, J. J. et al. Pathogen-induced inflammatory environment controls effector and memory CD8+T cell differentiation. J. Immunol. 187, 4967–4978 (2011).

    PubMed  PubMed Central  CAS  Google Scholar 

  42. 42.

    Mueller, S. N. et al. Qualitatively different memory CD8+T cells are generated after lymphocytic choriomeningitis virus and influenza virus infections. J. Immunol. 185, 2182–2190 (2010).

    PubMed  CAS  Google Scholar 

  43. 43.

    Martin, M. et al. Phenotypic and functional alterations in circulating memory CD8+ T cells with time after primary infection. PLOS Pathog. 11, e1005219 (2015).

    PubMed  PubMed Central  Google Scholar 

  44. 44.

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

    PubMed  CAS  Google Scholar 

  45. 45.

    Beura, L. K. et al. Intravital mucosal imaging of CD8+ resident memory T cells shows tissue-autonomous recall responses that amplify secondary memory. Nat. Immunol. 19, 173–182 (2018).

    PubMed  PubMed Central  CAS  Google Scholar 

  46. 46.

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

    PubMed  PubMed Central  CAS  Google Scholar 

  47. 47.

    Baenziger, J., Hengartner, H., Zinkernagel, R. M. & Cole, G. A. Induction or prevention of immunopathological disease by cloned cytotoxic T cell lines specific for lymphocytic choriomeningitis virus. Eur. J. Immunol. 16, 387–393 (1986).

    PubMed  CAS  Google Scholar 

  48. 48.

    Petito, C. K. & Adkins, B. Choroid plexus selectively accumulates T lymphocytes in normal controls and after peripheral immune activation. J. Neuroimmunol. 162, 19–27 (2005).

    PubMed  CAS  Google Scholar 

  49. 49.

    Mueller, S. N. & Mackay, L. K. Tissue-resident memory T cells: local specialists in immune defence. Nat. Rev. Immunol. 16, 79–89 (2015).

    PubMed  Google Scholar 

  50. 50.

    Ludwig, A. & Mentlein, R. Glial cross-talk by transmembrane chemokines CX3CL1 and CXCL16. J. Neuroimmunol. 198, 92–97 (2008).

    PubMed  CAS  Google Scholar 

  51. 51.

    Shan, Q. et al. The transcription factor Runx3 guards cytotoxic CD8+ effector T cells against deviation towards follicular helper T cell lineage. Nat. Immunol. 18, 931–939 (2017).

    PubMed  PubMed Central  CAS  Google Scholar 

  52. 52.

    Badovinac, V. P., Messingham, K. A. N., Jabbari, A., Haring, J. S. & Harty, J. T. Accelerated CD8+ T cell memory and prime-boost response after dendritic-cell vaccination. Nat. Med. 11, 748–756 (2005).

    PubMed  CAS  Google Scholar 

  53. 53.

    Manglani, M., Gossa, S. & McGavern, D. B. Leukocyte isolation from brain, spinal cord, and meninges for flow cytometric analysis. Curr. Prot. Immunol. 121, e44 (2018).

    Google Scholar 

  54. 54.

    Bendall, S. C. et al. Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 332, 687–696 (2011).

    PubMed  PubMed Central  CAS  Google Scholar 

  55. 55.

    Manglani, M. & McGavern, D. B. Intravital imaging of neuroimmune interactions through a thinned skull. Curr. Prot. Immunol. 120, 24.2.1–24.2.12 (2018).

    Google Scholar 

  56. 56.

    Hickman, H. D. Imaging CD8+ T cells during diverse viral infections. IntraVital 4, e1055425 (2015).

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank S. Perlman for critical review and comments on the manuscript and S. Anthony for helpful discussion. We thank J. Fishbaugh, H. Vignes and M. Shey (University of Iowa Flow Cytometry Core Facility) for cell sorting, I. Antoshechkin (California Institute of Technology) and Admera health for RNA-seq. Data herein were obtained from the Flow Cytometry Facility, which is a Carver College of Medicine Core Research Facilities/Holden Comprehensive Cancer Center Core Laboratory at the University of Iowa. This work was supported by grants from the National Institutes of Health (AI42767 to J.T.H., AI114543 to J.T.H. and V.P.B., GM134880 to V.P.B., AI121080 and AI139874 to H.-H.X., T32 AI007343 to S.L.U. and T32 AI007511 to I.J.J.) and the Veteran Affairs BLR&D Merit Review Program (BX002903) to H.-H.X.

Author information

Affiliations

Authors

Contributions

S.L.U. and J.T.H. designed experiments; S.L.U. conducted experiments; S.L.U. and I.J.J. analyzed data; I.J.J., Q.S. and L.L.P. provided technical assistance; V.P.B. and H.-H.X. provided essential reagents and intellectual input and S.L.U. and J.T.H. wrote the manuscript.

Corresponding author

Correspondence to John T. Harty.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Editor recognition statement L. A. Dempsey was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Enrichment of antigen specific CD8+ T-cells in the CNS after peripheral immunizations.

a, Gating strategy used to identify CD8+ T cell populations isolated from the CNS. Representative of 3 independent experiments with 4 mice per group.b-e, The proportion of donor OT-I, P14, or OVA tetramer+ cells of live CD8+ T-cells isolated from the Spleen (SP) peripheral blood (PBL) IV+ brain (IV+) or IV brain (IV) are graphed after infection with LCMV IV, p values (top to bottom) **** p<0.0001, **** p<0.0001, ****, p<0.0001, ** p=0.0038, and * p=0.0252 (b), DC-OVA-rLM-OVA prime boost IV, p values (top to bottom) **** p<0.0001, **** p<0.0001, *** p=0.0002, *** p=0.0.0004, and *** p=0.0005 (c), DC-GP33-rLM-GP33 prime boost IV, p values (top to bottom) *** p=0.0003, *** p=0.0002, and ****p<0.0001 (d), DC-OVA-VACV-OVA IV, p values (top to bottom) ** p=0.0014, ** p=0.0019, and * p=0.0201 (e). Data represent from 2 independent experiments with 3 mice per group. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Statistical significance was determined by One-way ANOVA with Tukey’s multiple comparisons test across all the groups using graphpad prism. f, NIH Swiss Webster mice were infected with rLM-OVA IV and the proportion of live CD8+, IV CD8+ and number of IV CD8+ cells in the CNS are graphed. Data pooled from 2 independent experiments for a total of 10 mice per group. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Two-tailed unpaired students T-test were determined using graphpad prism and p values (left to right) * p=0.0458, ** p=0.0048, and * p=0.0216.

Source data

Extended Data Fig. 2 Runx3 but not CD103 are required for generation of peripherally induced CNS Trm cells.

a,b, RNA-Seq analysis of memory OT-I cells isolated from SP, IV+, or IV brains of DC-OVA-rLM-OVA prime boosted mice. Fold change of IV+ vs IV OT-I cells of corresponding genes identified in Figure 2d (a). Fold change of IV+ vs IV OT-I cells of 40 different transcription factors associated with Trm or memory CD8+ T cell responses with selected genes indicated (b). RNA samples were isolated from 5 mice pooled per group in duplicate. c, CD103KO and WT mice were DC-OVA-rLM-OVA prime-boosted and brains were harvested at a memory time point. The proportion and numbers of ova-specific CD69+ IV CD8+ T-cells are plotted. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 individual experiments with 3 mice per group. d,e, WT or CD103KO P14 cells from naïve donors were adoptively transferred into WT CD45.1 hosts prior to LCMV infection IP. At a memory time point, proportion of donor P14 cells were determined and are depicted for WT P14 cells (d) and CD103KO P14 cells (e). Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 individual experiments with 5 mice per group. Statistical significance was determined by One-way ANOVA with Tukey’s multiple comparisons test across all the groups using graphpad prism with p values (top to bottom) **** p<0.0001, **** p<0.00001, and **** p<0.0001 (d) and ** p=0.0025, ** p=0.0026, and ** p=0.0033 (e). f,g, WT or Runx3KO P14 cells were adoptively transferred into WT CD45.1 hosts prior to LCMV infection IP. At a memory time point, proportion of donor P14 cells were determined and are depicted for WT P14 cells (f) and Runx3KO P14 cells (g). Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 individual experiments with 5 mice per group. Statistical significance was determined by One-way ANOVA with Tukey’s multiple comparisons test across all the groups using graphpad prism with p values (top to bottom) * p=0.0159, and * p=0.0279 (f).

Source data

Extended Data Fig. 3 Increased representation of antigen specific CD8+ T-cells after peripheral immunizations is CNS specific.

a-h, Thy1.1 eGFP+ OT-I T-cells were transferred into naïve mice that were DC-OVA-rLM-OVA prime boosted. Representative flow plots (a) showing proportion of OT-I cells of IV+ or IV Thy1.1 CD8+ T-cells in each organ and cumulative proportion of OT-I cells in each organ, p values (left to right) ** p=0.0028 (b). Proportion of OT-I cells expressing CD103, p values (left to right) *** p=0.00371, * p=0.0100 (c) and CX3CR1, p values (left to right) *** p=0.0001, ** p=0.0027 (d) in IV+ vs IV brain and meninges. gMFI of CD69, p values (left to right) *** p=0.0005, ** p=0.0013 (e), CXCR6, p values (left to right) **** p<0.0001, *** p=0.0004 (f), and CD49a, p values (left to right) **** p<0.0001, ** p=0.0025 (g) of OT-I cells from IV+ vs IV brain and meninges are shown. SPADE analysis of OT-I cells from each organ are depicted (h). Markers used to distinguish populations include CD8α, CD11a, CD44, CD49a, CD69, CD103, CXCR6, and CX3CR1. Data are representative of 2 independent experiments with 3 mice per group. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Two-tailed unpaired l students T-test were determined using graphpad prism.

Source data

Extended Data Fig. 4 Peripherally induced CNS Trm cells are resistant to systemic depletion.

a,b, Thy1.1 eGFP+ OT-I T-cells were transferred into naïve mice that were DC-OVA-rLM-OVA prime boosted. At a memory time point, mice were control treated (PBS) or treated with 2, 5, or 10 μg of anti-Thy1.1 Ab IP. One week after depletion, proportions and phenotype of OT-I cells were determined. Representative flow plots gated on live CD8+ cells from Spleen, PBL and IV+ or IV brain showing proportions of OT-I cells (a) and gMFI of CD69 gated on IV OT-I cells (b). Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 individual experiments with 3 mice per group. Statistical significance was determined by One-way ANOVA with Tukey’s multiple comparisons test across all the groups using graphpad prism with p values (top to bottom) *** p=0.0005, *** p=0.0004, and * p=0.0144. c, Thy1.1 eGFP+ OT-I T-cells were transferred into naïve mice that were DC-OVA-rLM-OVA prime boosted. After memory formation, mice were control treated (PBS) or treated with 2 μg of anti-Thy1.1 Ab IP. One week after depletion, OT-I cells in the CNS were imaged and maximum speed was determined. Graph depicts mean with each dot representing an individual OT-I cell from 71 cells from 12 individual movies (PBS), and 103 cells from 16 individual movies (2 μg). Statistical significance was determined by two-tailed Mann Whitney test using graphpad prism with **** p<0.0001.

Source data

Extended Data Fig. 5 Enhanced recall response of peripherally induced brain Trm cells is specific to CNS.

a,b, OT-I T-cells were transferred into recipient mice that were DC-OVA-VACV-OVA prime boosted. At a memory time point, mice were either unchallenged or challenged with rLM-OVA IC and organs were harvested 2 days later and stained for CD25 and intracellular IFN-γ directly ex vivo. Proportion of CD25+, p value **** p<0.0001 (a) and IFN-γ+, p value *** p=0.0004 (b) OT-I cells from each organ are graphed. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 independent experiments with 4 mice per group. Statistical significance was determined by two-tailed unpaired students T-test for each organ using graphpad prism. c-e, P14 T-cells were transferred into recipient mice that were immunized with LCMV IP. At a memory time point, mice were treated with PBS or 2 μg anti-Thy1.1 Ab IP. One week post depletion, mice were challenged with rLM-GP33 IC and organs were harvested 2 days later and stained for CD25 and intracellular IFN-γ directly ex vivo. Proportion of CD25+, p values (left to right) ** p=0.0099, ** p=0.0014 (c), IFN-γ+, p values (left to right) * p=0.0308, ** p=0.0069 (d) and number of IV P14 cells (e) from the brain are graphed. Graphs show the mean +/- standard deviation with each dot representing an individual mouse. Data are representative of 2 independent experiments with 3 mice per group. Two-tailed unpaired students T-test was determined using graphpad prism.

Source data

Extended Data Fig. 6 Peripherally induced brain resident CD8+ T-cells mediate protection against CNS infections.

a, Thy1.1 OT-I cells were transferred into mice and DC-OVA-rLM-OVA prime boosted. After memory formation, these mice and naïve controls were challenged with VSV-OVA IN. Virus titers in brains were determined at day 3 post infection. Data are combined from 2 independent experiments for a total of 10 mice per group. Graph shows the mean +/- standard deviation with each dot representing an individual mouse. Two-tailed unpaired students T-test was determined using graphpad prism where ** p=0.0021. b, Thy1.1 OT-I cells were transferred into recipient mice that were DC-OVA-rLM-OVA prime boosted. After memory formation, mice were treated with PBS or 2 μg anti-Thy1.1 Ab IP. One week after depletion these mice and naïve controls mice were challenged with VSV-OVA IC. Kaplan Meier survival curves depicted. Data from representative experiment with 6 (black), 8 (red), and 9 (blue) mice per group (top to bottom). Graphpad prism used to determine significance using Mantel-Cox test for each group comparing to the Naïve + IC challenge group with p values * p=0.014 (red), and ** p=0.0056 (blue). c,d, Thy1.1 OT-I or P14 cells were transferred into recipient mice that were DC-OVA-rLM-OVA or DC-GP33-rLM-GP33 prime boosted, respectively. After memory formation, mice were treated with PBS or 2 μg anti-Thy1.1 Ab IP. Frequency of OVA or GP33 tetramer positive cells and proportion of transgenic OT-I or P14 T-cells were determined prior to and 5 days post depletion. Representative flow plots (c) and cumulative data (d) of 2 independent experiments with 5 mice per group are shown.

Source data

Supplementary information

Reporting Summary

Supplementary Video 1

Behavior of peripherally induced brain TRM cells. Representative time-lapse video shows dynamics of memory CD8+ T cells through a thinned skull by two-photon laser scanning microscopy of DC-OVA-rLM-OVA immunized mice. eGFP OT-I cells (green) and blood vessels (red) show projections representing z-stacks 80 μm in depth imaged over a 30-min time period. One representative video is shown from 12 movies. Please see Supplementary Video 2 for a second representative video.

Supplementary Video 2

Behavior of peripherally induced brain TRM cells. Representative time-lapse video shows dynamics of memory CD8+ T cells through a thinned skull by two-photon laser scanning microscopy of DC-OVA-rLM-OVA immunized mice. eGFP OT-I cells (green) and blood vessels (red) show projections representing z-stacks 80 μm in depth imaged over a 30-min time period. One representative video is shown from 12 movies. Please see Supplementary Video 1 for a second representative video.

Supplementary Video 3

Behavior of peripherally induced brain TRM cells. Representative time-lapse video shows dynamics of memory CD8+ T-cells through a thinned skull by two-photon laser scanning microscopy of DC-OVA-rLM-OVA immunized mice. eGFP OT-I cells (green), Thy1.1 PE+ OT-I cells (pink) and blood vessels (red) show projections representing z-stacks 80 μm in depth imaged over a 30-min time period. Representative video and cell tracks of eGFP+ cells are depicted. Representative videos are shown from 15 movies.

Supplementary Video 4

Behavior of peripherally induced brain TRM cells. Representative time-lapse video shows dynamics of memory CD8+ T cells through a thinned skull by two-photon laser scanning microscopy of DC-OVA-rLM-OVA immunized mice. eGFP OT-I cells (green), Thy1.1 PE+ OT-I cells (pink) and blood vessels (red) show projections representing z-stacks 80 μm in depth imaged over a 30-min time period. Representative video and cell tracks of Thy1.1+ cells are depicted. Companion video to Supplementary Video 3. Representative video is shown from 15 movies.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 5

Statistical source data.

Source Data Fig. 6

Statistical source data.

Source Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 1

Statistical source data.

Source Data Extended Data Fig. 2

Statistical source data.

Source Data Extended Data Fig. 3

Statistical source data.

Source Data Extended Data Fig. 4

Statistical source data.

Source Data Extended Data Fig. 5

Statistical source data.

Source Data Extended Data Fig. 6

Statistical source data.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Urban, S.L., Jensen, I.J., Shan, Q. et al. Peripherally induced brain tissue–resident memory CD8+ T cells mediate protection against CNS infection. Nat Immunol 21, 938–949 (2020). https://doi.org/10.1038/s41590-020-0711-8

Download citation

Further reading

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