Tissue-resident memory T cells (TRMs) are a novel nonvascular memory T cell subset. Although CD8+ TRMs are well-characterized, CD4+ TRMs—especially lung-resident memory Th17 cells—are still being defined. In this study, we characterized lung-resident memory Th17 cells (lung TRM17) and their role in protection against the highly virulent fungus Cryptococcus gattii. We found that intravenously transferred DCs preferentially migrated to lungs and attracted recipient DCs and led to the induction of long-lived Th17 cells expressing characteristic markers. This population could be clearly discriminated from circulating T cells by intravascular staining and was not depleted by the immunosuppressive agent FTY720. The C. gattii antigen re-stimulation assay revealed that vaccine-induced lung Th17 cells produced IL-17A but not IFNγ. The DC vaccine significantly increased IL-17A production and suppressed fungal burden in the lungs and improved the survival of mice infected with C. gattii. This protective effect was significantly reduced in the IL-17A knockout (KO) mice, but not in the FTY720-treated mice. The protective effect also coincided with the activation of neutrophils and multinucleated giant cells, and these inflammatory responses were suppressed in the vaccinated IL-17A KO mice. Overall, these data demonstrated that the systemic DC vaccine induced lung TRM17, which played a substantial role in anti-fungal immunity.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $92.33 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Mueller, S. N. & Mackay, L. K. Tissue-resident memory T cells: local specialists in immune defence. Nat. Rev. Immunol. 16, 79–89 (2016).
Park, C. O. & Kupper, T. S. The emerging role of resident memory T cells in protective immunity and inflammatory disease. Nat. Med. 21, 688–697 (2015).
Ganesan, A.-P. et al. Tissue-resident memory features are linked to the magnitude of cytotoxic T cell responses in human lung cancer. Nat. Immunol. 18, 940–950 (2017).
Hayashizaki, K. et al. Myosin light chains 9 and 12 are functional ligands for CD69 that regulate airway inflammation. Sci. Immunol. 1, eaaf9154 (2016).
Mackay, L. K. et al. Cutting edge: CD69 interference with sphingosine-1-phosphate receptor function regulates peripheral T cell retention. J. Immunol. 194, 2059–2063 (2015).
Lin, Y., Slight, S. R. & Khader, S. A. Th17 cytokines and vaccine-induced immunity. Semin. Immunopathol. 32, 79–90 (2010).
Ueno, K. et al. Dendritic cell-based immunization ameliorates pulmonary infection with highly virulent Cryptococcus gattii. Infect. Immun. 83, 1577–1586 (2015).
Lizarazo, J. et al. Retrospective study of the epidemiology and clinical manifestations of Cryptococcus gattii infections in Colombia from 1997–2011. PLoS Negl. Trop. Dis. 8, e3272 (2014).
CDC Emergence of Cryptococcus gattii, Pacific Northwest, 2004–2010. Morb. Mortal. Wkly. Rep. 59, 865–868 (2010).
Palucka, K. & Banchereau, J. Dendritic-cell-based therapeutic cancer vaccines. Immunity 39, 38–48 (2013).
García, F. et al. A therapeutic dendritic cell-based vaccine for HIV-1 infection. J. Infect. Dis. 203, 473–478 (2011).
Roy, R. M. & Klein, B. S. Dendritic cells in antifungal immunity and vaccine design. Cell Host Microbe 11, 436–446 (2012).
Turner, D. L. & Farber, D. L. Mucosal resident memory CD4 T cells in protection and immunopathology. Front. Immunol. 5, 331 (2014).
McKinstry, K. K. et al. Effector CD4 T-cell transition to memory requires late cognate interactions that induce autocrine IL-2. Nat. Commun. 5, 5377 (2014).
Takenaka, M. et al. Antibodies to MHC class II molecules induce autoimmunity: critical role for macrophages in the immunopathogenesis of obliterative airway disease. PLoS ONE 7, e42370 (2012).
Anderson, K. G. et al. Intravascular staining for discrimination of vascular and tissue leukocytes. Nat. Protoc. 9, 209–222 (2014).
Turner, D. L. et al. Lung niches for the generation and maintenance of tissue-resident memory T cells. Mucosal Immunol. 7, 501–510 (2014).
Laidlaw, B. J. et al. CD4(+) T cell help guides formation of CD103(+) lung-resident memory CD8(+) T cells during influenza viral infection. Immunity 41, 633–645 (2014).
Sionov, E. et al. Type I IFN induction via poly-ICLC protects mice against Cryptococcosis. PLoS Pathog. 11, e1005040 (2015).
Iwata, A. et al. Th2-type inflammation instructs inflammatory dendritic cells to induce airway hyperreactivity. Int. Immunol. 26, 103–114 (2013).
Lewis, S. M. et al. Expression of CD11c and EMR2 on neutrophils: potential diagnostic biomarkers for sepsis and systemic inflammation. Clin. Exp. Immunol. 182, 184–194 (2015).
Villa-Ambriz, J., Rodríguez-Orozco, A. R., Béjar-Lozano, C. & Cortés-Rojo, C. The increased expression of CD11c and CD103 molecules in the neutrophils of the peripheral blood treated with a formula of bacterial ribosomes and proteoglycans of Klebsiella pneumoniae. Arch. Bronconeumol. 48, 316–319 (2012).
Percopo, C. M. et al. SiglecF+Gr1hi eosinophils are a distinct subpopulation within the lungs of allergen-challenged mice. J. Leukoc. Biol. 101, 321–328 (2017).
Kiwamoto, T., Kawasaki, N., Paulson, J. C. & Bochner, B. S. Siglec-8 as a drugable target to treat eosinophil and mast cell-associated conditions. Pharmacol. Ther. 135, 327–336 (2012).
Ishizuka, M. et al. Interleukin-17A and neutrophils in a murine model of bird-related hypersensitivity pneumonitis. PLoS ONE 10, e0137978 (2015).
Zhang, M. et al. Defining the in vivo function of Siglec-F, a CD33-related Siglec expressed on mouse eosinophils. Blood 109, 4280–4287 (2007).
Yoshioka, Y. et al. Neutrophils and the S100A9 protein critically regulate granuloma formation. Blood Adv. 1, 184–192 (2016).
Gopal, R. et al. Interleukin-17-dependent CXCL-13 mediates mucosal vaccine-induced immunity against tuberculosis. Mucosal Immunol. 6, 972–984 (2013).
Murdock, B. J., Huffnagle, G. B., Olszewski, M. A. & Osterholzer, J. J. Interleukin-17A enhances host defense against cryptococcal lung infection through effects mediated by leukocyte recruitment, activation, and gamma interferon production. Infect. Immun. 82, 937–948 (2014).
Khan, T. N., Mooster, J. L., Kilgore, A. M., Osborn, J. F. & Nolz, J. C. Local antigen in nonlymphoid tissue promotes resident memory CD8+ T cell formation during viral infection. J. Exp. Med. 213, 951–966 (2016).
Pepper, M. et al. Different routes of bacterial infection induce long-lived TH1 memory cells and short-lived TH17 cells. Nat. Immunol. 11, 83–89 (2009).
Christensen, D., Mortensen, R., Rosenkrands, I., Dietrich, J. & Andersen, P. Vaccine-induced Th17 cells are established as resident memory cells in the lung and promote local IgA responses. Mucosal Immunol. 10, 260–270 (2017).
Shimizu, K. et al. KLRG+ invariant natural killer T cells are long-lived effectors. Proc. Natl. Acad. Sci. USA 111, 12474–12479 (2014).
Lappin, M. B. et al. Analysis of mouse dendritic cell migration in vivo upon subcutaneous and intravenous injection. Immunology 98, 181–188 (1999).
Mullins, D. W. et al. Route of immunization with peptide-pulsed dendritic cells controls the distribution of memory and effector T cells in lymphoid tissues and determines the pattern of regional tumor control. J. Exp. Med. 198, 1023–1034 (2003).
Sheasley-O’Neill, S. L., Brinkman, C. C., Ferguson, A. R., Dispenza, M. C. & Engelhard, V. H. Dendritic cell immunization route determines integrin expression and lymphoid and nonlymphoid tissue distribution of CD8 T cells. J. Immunol. 178, 1512–1522 (2007).
Kryczek, I. et al. Human TH17 cells are long-lived effector memory cells. Sci. Transl. Med. 3, 104ra100 (2011).
Sercan Alp, Ö. et al. Memory CD8(+) T cells colocalize with IL-7(+) stromal cells in bone marrow and rest in terms of proliferation and transcription. Eur. J. Immunol. 45, 975–987 (2015).
Xu, X. et al. Autophagy is essential for effector CD8(+) T cell survival and memory formation. Nat. Immunol. 15, 1152–1161 (2014).
Araki, K. et al. mTOR regulates memory CD8 T-cell differentiation. Nature 460, 108–112 (2009).
Muranski, P. et al. Th17 cells are long lived and retain a stem cell-like molecular signature. Immunity 35, 972–985 (2011).
Maekawa, Y. et al. Notch controls the survival of memory CD4+ T cells by regulating glucose uptake. Nat. Med. 21, 55–61 (2015).
Pan, Y. et al. Survival of tissue-resident memory T cells requires exogenous lipid uptake and metabolism. Nature 543, 252–256 (2017).
Reya, T. et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 423, 409–414 (2003).
Gattinoni, L. et al. Wnt signaling arrests effector T cell differentiation and generates CD8+ memory stem cells. Nat. Med. 15, 808–813 (2009).
Iijima, N. & Iwasaki, A. T cell memory. A local macrophage chemokine network sustains protective tissue-resident memory CD4 T cells. Science 346, 93–98 (2014).
Kondrack, R. M. et al. Interleukin 7 regulates the survival and generation of memory CD4 cells. J. Exp. Med. 198, 1797–1806 (2003).
Glennie, N. D., Volk, S. W. & Scott, P. Skin-resident CD4+ T cells protect against Leishmania major by recruiting and activating inflammatory monocytes. PLoS Pathog. 13, e1006349 (2017).
Nakae, S. et al. Antigen-specific T cell sensitization is impaired in IL-17-deficient mice, causing suppression of allergic cellular and humoral responses. Immunity 17, 375–387 (2002).
Adachi, Y. et al. Distinct germinal center selection at local sites shapes memory B cell response to viral escape. J. Exp. Med. 212, 1709–1723 (2015).
This work was supported by KAKENHI (15K21644, 16H05349, and 17K18385) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, by grants from the Japan Agency for Medical Research and Development, AMED, by the Takeda Science Foundation, and by the LEGEND Research Grant Program 2015 from Tomy Digital Biology Co. Ltd. We also thank Drs. Yu Adachi, Taishi Onodera, Yoshimasa Takahashi, and Koji Hayashizaki who provided the technical advice for immunohistochemical analysis.
The authors declare no competing interests.
All animal experiment protocols were approved by the Ethical Committee of the National Institute of Infectious Diseases, Japan (approval numbers 115015, 116019, 116124, 215031, 215036, 215047, 114029, 117032, and 114029) and were performed in accordance with the approved guidelines and regulations.
Electronic supplementary material
About this article
Medical Mycology (2019)