Abstract
Leukocytes must traverse inflamed tissues to effectively control local infection. Although motility in dense tissues seems to be integrin independent and based on actomyosin-mediated protrusion and contraction, during inflammation, changes to the extracellular matrix (ECM) may necessitate distinct motility requirements. Indeed, we found that the interstitial motility of T cells was critically dependent on Arg-Gly-Asp (RGD)-binding integrins in the inflamed dermis. Inflammation-induced deposition of fibronectin was functionally linked to higher expression of integrin αV on effector CD4+ T cells. By intravital multiphoton imaging, we found that the motility of CD4+ T cells was dependent on αV expression. Selective blockade or knockdown of αV arrested T helper type 1 (TH1) cells in the inflamed tissue and attenuated local effector function. Our data demonstrate context-dependent specificity of lymphocyte movement in inflamed tissues that is essential for protective immunity.
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References
Nourshargh, S., Hordijk, P.L. & Sixt, M. Breaching multiple barriers: leukocyte motility through venular walls and the interstitium. Nat. Rev. Mol. Cell Biol. 11, 366–378 (2010).
Friedl, P. & Weigelin, B. Interstitial leukocyte migration and immune function. Nat. Immunol. 9, 960–969 (2008).
Friedl, P., Entschladen, F., Conrad, C., Niggemann, B. & Zanker, K.S. CD4+ T lymphocytes migrating in three-dimensional collagen lattices lack focal adhesions and utilize β1 integrin-independent strategies for polarization, interaction with collagen fibers and locomotion. Eur. J. Immunol. 28, 2331–2343 (1998).
Jacobelli, J. et al. Confinement-optimized three-dimensional T cell amoeboid motility is modulated via myosin IIA-regulated adhesions. Nat. Immunol. 11, 953–961 (2010).
Woolf, E. et al. Lymph node chemokines promote sustained T lymphocyte motility without triggering stable integrin adhesiveness in the absence of shear forces. Nat. Immunol. 8, 1076–1085 (2007).
Lämmermann, T. et al. Rapid leukocyte migration by integrin-independent flowing and squeezing. Nature 453, 51–55 (2008).
Sorokin, L. The impact of the extracellular matrix on inflammation. Nat. Rev. Immunol. 10, 712–723 (2010).
Sigmundsdottir, H. & Butcher, E.C. Environmental cues, dendritic cells and the programming of tissue-selective lymphocyte trafficking. Nat. Immunol. 9, 981–987 (2008).
Ray, S.J. et al. The collagen binding α1β1 integrin VLA-1 regulates CD8 T cell-mediated immune protection against heterologous influenza infection. Immunity 20, 167–179 (2004).
Okada, T. Two-photon microscopy analysis of leukocyte trafficking and motility. Semin. Immunopathol. 32, 215–225 (2010).
Schumann, K. et al. Immobilized chemokine fields and soluble chemokine gradients cooperatively shape migration patterns of dendritic cells. Immunity 32, 703–713 (2010).
Bajénoff, M. et al. Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes. Immunity 25, 989–1001 (2006).
Wilson, E.H. et al. Behavior of parasite-specific effector CD8+ T cells in the brain and visualization of a kinesis-associated system of reticular fibers. Immunity 30, 300–311 (2009).
Matheu, M.P. et al. Imaging of effector memory T cells during a delayed-type hypersensitivity reaction and suppression by Kv1.3 channel block. Immunity 29, 602–614 (2008).
Boissonnas, A., Fetler, L., Zeelenberg, I.S., Hugues, S. & Amigorena, S. In vivo imaging of cytotoxic T cell infiltration and elimination of a solid tumor. J. Exp. Med. 204, 345–356 (2007).
Mrass, P. et al. Random migration precedes stable target cell interactions of tumor-infiltrating T cells. J. Exp. Med. 203, 2749–2761 (2006).
Werr, J., Xie, X., Hedqvist, P., Ruoslahti, E. & Lindbom, L. β1 integrins are critically involved in neutrophil locomotion in extravascular tissue In vivo. J. Exp. Med. 187, 2091–2096 (1998).
Gray, E.E., Suzuki, K. & Cyster, J.G. Cutting edge: Identification of a motile IL-17-producing gammadelta T cell population in the dermis. J. Immunol. 186, 6091–6095 (2011).
Sumaria, N. et al. Cutaneous immunosurveillance by self-renewing dermal gammadelta T cells. J. Exp. Med. 208, 505–518 (2011).
Wang, Q. et al. CD4 promotes breadth in the TCR repertoire. J. Immunol. 167, 4311–4320 (2001).
Filipe-Santos, O. et al. A dynamic map of antigen recognition by CD4 T cells at the site of Leishmania major infection. Cell Host Microbe 6, 23–33 (2009).
Egawa, G. et al. In vivo imaging of T-cell motility in the elicitation phase of contact hypersensitivity using two-photon microscopy. J. Invest. Dermatol. 131, 977–979 (2011).
Stetson, D.B. et al. Constitutive cytokine mRNAs mark natural killer (NK) and NK T cells poised for rapid effector function. J. Exp. Med. 198, 1069–1076 (2003).
Ng, L.G. et al. Migratory dermal dendritic cells act as rapid sensors of protozoan parasites. PLoS Pathog. 4, e1000222 (2008).
Springer, T.A. Adhesion receptors of the immune system. Nature 346, 425–434 (1990).
Campbell, I.D. & Humphries, M.J. Integrin structure, activation, and interactions. Cold Spring Harb. Perspect. Biol. 3, 1–14 (2011).
DeNucci, C.C. & Shimizu, Y. β1 integrin is critical for the maintenance of antigen-specific CD4 T cells in the bone marrow but not long-term immunological memory. J. Immunol. 186, 4019–4026 (2011).
Ruoslahti, E. RGD and other recognition sequences for integrins. Annu. Rev. Cell Dev. Biol. 12, 697–715 (1996).
Clark, R.A., Dvorak, H.F. & Colvin, R.B. Fibronectin in delayed-type hypersensitivity skin reactions: associations with vessel permeability and endothelial cell activation. J. Immunol. 126, 787–793 (1981).
Clark, R.A. et al. Fibronectin deposition in delayed-type hypersensitivity. Reactions of normals and a patient with afibrinogenemia. J. Clin. Invest. 74, 1011–1016 (1984).
Kusubata, M. et al. Spatiotemporal changes of fibronectin, tenascin-C, fibulin-1, and fibulin-2 in the skin during the development of chronic contact dermatitis. J. Invest. Dermatol. 113, 906–912 (1999).
Klebe, R.J. Isolation of a collagen-dependent cell attachment factor. Nature 250, 248–251 (1974).
Richter, M. et al. Collagen distribution and expression of collagen-binding α1β1 (VLA-1) and α2β1 (VLA-2) integrins on CD4 and CD8 T cells during influenza infection. J. Immunol. 178, 4506–4516 (2007).
Campbell, D.J. & Butcher, E.C. Rapid acquisition of tissue-specific homing phenotypes by CD4(+) T cells activated in cutaneous or mucosal lymphoid tissues. J. Exp. Med. 195, 135–141 (2002).
Sigmundsdottir, H. et al. DCs metabolize sunlight-induced vitamin D3 to 'program' T cell attraction to the epidermal chemokine CCL27. Nat. Immunol. 8, 285–293 (2007).
Fazilleau, N., McHeyzer-Williams, L.J., Rosen, H. & McHeyzer-Williams, M.G. The function of follicular helper T cells is regulated by the strength of T cell antigen receptor binding. Nat. Immunol. 10, 375–384 (2009).
McLachlan, J.B., Catron, D.M., Moon, J.J. & Jenkins, M.K. Dendritic cell antigen presentation drives simultaneous cytokine production by effector and regulatory T cells in inflamed skin. Immunity 30, 277–288 (2009).
Sojka, D.K. & Fowell, D.J. Regulatory T cells inhibit acute IFN-γ synthesis without blocking T-helper cell type 1 (Th1) differentiation via a compartmentalized requirement for IL-10. Proc. Natl. Acad. Sci. USA 108, 18336–18341 (2011).
Renkawitz, J. et al. Adaptive force transmission in amoeboid cell migration. Nat. Cell Biol. 11, 1438–1443 (2009).
Friedl, P. & Wolf, K. Plasticity of cell migration: a multiscale tuning model. J. Cell Biol. 188, 11–19 (2010).
Sandig, H. et al. Fibronectin is a TH1-specific molecule in human subjects. J Allergy Clin Immunol 124, 528–535 (2009).
Shulman, Z. et al. Lymphocyte crawling and transendothelial migration require chemokine triggering of high-affinity LFA-1 integrin. Immunity 30, 384–396 (2009).
Park, E.J. et al. Distinct roles for LFA-1 affinity regulation during T-cell adhesion, diapedesis, and interstitial migration in lymph nodes. Blood 115, 1572–1581 (2010).
Rutkowski, J.M. & Swartz, M.A. A driving force for change: interstitial flow as a morphoregulator. Trends Cell Biol. 17, 44–50 (2007).
Conrad, C. et al. α1β1 integrin is crucial for accumulation of epidermal T cells and the development of psoriasis. Nat. Med. 13, 836–842 (2007).
Yang, Z. et al. Absence of integrin-mediated TGFβ1 activation in vivo recapitulates the phenotype of TGFβ1-null mice. J. Cell Biol. 176, 787–793 (2007).
Luzina, I.G. et al. Regulation of pulmonary inflammation and fibrosis through expression of integrins αVβ3 and αVβ5 on pulmonary T lymphocytes. Arthritis Rheum. 60, 1530–1539 (2009).
Acharya, M. et al. αV Integrin expression by DCs is required for Th17 cell differentiation and development of experimental autoimmune encephalomyelitis in mice. J. Clin. Invest. 120, 4445–4452 (2010).
Païdassi, H. et al. Preferential expression of integrin αVβ8 promotes generation of regulatory T cells by mouse CD103+ dendritic cells. Gastroenterology 141, 1813–1820 (2011).
Masuoka, M. et al. Periostin promotes chronic allergic inflammation in response to Th2 cytokines. J. Clin. Invest. 122, 2590–2600 (2012).
Kudo, M. et al. IL-17A produced by αβ T cells drives airway hyper-responsiveness in mice and enhances mouse and human airway smooth muscle contraction. Nat. Med. 18, 547–554 (2012).
Lacy-Hulbert, A. et al. Ulcerative colitis and autoimmunity induced by loss of myeloid αV integrins. Proc. Natl. Acad. Sci. USA 104, 15823–15828 (2007).
Yang, J.T. & Hynes, R.O. Fibronectin receptor functions in embryonic cells deficient in α5β1 integrin can be replaced by αV integrins. Mol. Biol. Cell 7, 1737–1748 (1996).
van der Flier, A. et al. Endothelial α5 and αV integrins cooperate in remodeling of the vasculature during development. Development 137, 2439–2449 (2010).
Wennerberg, K. et al. β1 integrin-dependent and -independent polymerization of fibronectin. J. Cell Biol. 132, 227–238 (1996).
Acknowledgements
We thank N. Killeen (University of California, San Francisco) for WT15 mice; R. Locksley (University of California, San Francisco) for IFN-γ reporter mice (Yeti mice); M. Nussenzweig (The Rockefeller University) for CD11c-YFP mice; R. Hynes (MIT) for Itga5fl/fl mice; B. Leon and F. Lund (University of Alabama) for H. polygyrus–infected tissue; R. Germain and J. Egen for technical assistance; K. Kasischke, L. Callahan, E. Brown and the University of Rochester Medical Center Multiphoton Core for support; Kihong Lim for technical assistance; J. Miller and S. Georas for comments on the manuscript; and members of the Fowell and Kim laboratories for discussions and support. Supported by the US National Institutes of Health (AI072690 and AI088427 to D.J.F.; HL018208 and HL087088 to M.K.; and AI089079 to M.G.O.), the American Heart Association (11SDG7520018 to Y.-M.H.) and the intramural program of the National Institute of Allergy and Infectious Diseases (B.R.A. and M.M.-S.).
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M.G.O., A.G. and D.J.F. designed the study; M.G.O., A.G., K.L., M.A., A.C.B.-M., A.H. and Y.-M.H. did the experiments; M.G.O., A.G., B.R.A., A.F.R., M.M.-S. and D.J.F. analyzed the data; Y.-M.H., M.K., D.J.T., A.L.-H., H.Y. and M.M.-S. provided reagents and conceptual advice and M.G.O. and D.J.F. wrote the manuscript.
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Supplementary Figures 1–10 (PDF 15587 kb)
Th1 effector T cell motility in the CFA-inflamed dermis.
CFSE-labeled WT15 Th1 cells were transferred into naïve BALB/c mice that were then immunized in the left and right ear with 1 μg pLACK/CFA. Three days later, T cells were visualized in the ear dermis by intravital multiphoton microscopy. WT15 cells (green) are shown crawling within the tissue collagen matrix visualized by SHG (blue). The movie represents a two-dimensional z-projection time series of a 50 μm thick imaging volume. (MOV 5783 kb)
Arrest of effector T cell motility with a blocking antibody to β1 integrin.
Mice were treated as in Movie 1 and were imaged three days later. After approximately 40 or 32 minutes, blocking antibody to β1 integrin (anti-CD29, 100 μg) was injected intravenously and T cell motility was monitored. The movie represents two-dimensional z-projection time series of 50 μm thick imaging volumes. (MOV 4579 kb)
RGD peptides halt effector T cell motility.
Mice were treated as in Movie 1 and imaged three days later. Immediately prior to imaging RGD (right panel) or control RAD (left pane) peptide was injected directly into the ear dermis (50μg peptide/ear) (MOV 1673 kb)
T cell motility is inhibited by a blocking antibody to αv integrin.
Mice were treated as in Movie 1 and were imaged three days later. After approximately 20 minutes, blocking antibody to αv (100μg) was injected intravenously and T cell motility was monitored. The movie represents two-dimensional z-projection time series of 50μm thick imaging volumes. (MOV 9310 kb)
αv-dependency for Th1 cell interstitial motility.
Naïve β 1-/- OT-II cells were stimulated with peptide/APC under Th1 conditions. One day after stimulation cells were transduced with control MSCV vector or αv-shRNA. On day 5 of culture, cells were harvested (transduction efficiency, 30-40% GFP+) and cells transferred to naive albino B6 mice that were then immunized in the ear with 1 μg pOVA/CFA. Three days later, T cells were visualized in the ear dermis by intravital multiphoton microscopy. Control vector GFP+ cells (left panel) and αv-shRNA GFP+ cells (right panel). (MOV 599 kb)
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Overstreet, M., Gaylo, A., Angermann, B. et al. Inflammation-induced interstitial migration of effector CD4+ T cells is dependent on integrin αV. Nat Immunol 14, 949–958 (2013). https://doi.org/10.1038/ni.2682
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DOI: https://doi.org/10.1038/ni.2682
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