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Inflammation-induced interstitial migration of effector CD4+ T cells is dependent on integrin αV

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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Figure 1: Migratory patterns of effector TH1 T cells in CFA-inflamed ear dermis.
Figure 2: The migration of T cells is guided by ECM fibers.
Figure 3: RGD-dependent integrin blockade impairs T cell motility in the inflamed dermis.
Figure 4: CD4+ T cells use integrin αV for intradermal motility.
Figure 5: Broad expression of integrin αV by effector CD4+ T cells across tissues and types of inflammation.
Figure 6: 'Lymph node–emigrant' effector T cells have high expression of integrin αV.
Figure 7: Nonredundant role for integrin αV in the intradermal motility of effector CD4+ T cells and antimicrobial clearance.

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References

  1. 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).

    Article  CAS  PubMed  Google Scholar 

  2. Friedl, P. & Weigelin, B. Interstitial leukocyte migration and immune function. Nat. Immunol. 9, 960–969 (2008).

    CAS  PubMed  Google Scholar 

  3. 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).

    CAS  PubMed  Google Scholar 

  4. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 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).

    CAS  PubMed  Google Scholar 

  6. Lämmermann, T. et al. Rapid leukocyte migration by integrin-independent flowing and squeezing. Nature 453, 51–55 (2008).

    PubMed  Google Scholar 

  7. Sorokin, L. The impact of the extracellular matrix on inflammation. Nat. Rev. Immunol. 10, 712–723 (2010).

    CAS  PubMed  Google Scholar 

  8. Sigmundsdottir, H. & Butcher, E.C. Environmental cues, dendritic cells and the programming of tissue-selective lymphocyte trafficking. Nat. Immunol. 9, 981–987 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. 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).

    CAS  PubMed  Google Scholar 

  10. Okada, T. Two-photon microscopy analysis of leukocyte trafficking and motility. Semin. Immunopathol. 32, 215–225 (2010).

    PubMed  PubMed Central  Google Scholar 

  11. Schumann, K. et al. Immobilized chemokine fields and soluble chemokine gradients cooperatively shape migration patterns of dendritic cells. Immunity 32, 703–713 (2010).

    CAS  PubMed  Google Scholar 

  12. Bajénoff, M. et al. Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes. Immunity 25, 989–1001 (2006).

    PubMed  PubMed Central  Google Scholar 

  13. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Mrass, P. et al. Random migration precedes stable target cell interactions of tumor-infiltrating T cells. J. Exp. Med. 203, 2749–2761 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. 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).

    CAS  PubMed  Google Scholar 

  19. Sumaria, N. et al. Cutaneous immunosurveillance by self-renewing dermal gammadelta T cells. J. Exp. Med. 208, 505–518 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Wang, Q. et al. CD4 promotes breadth in the TCR repertoire. J. Immunol. 167, 4311–4320 (2001).

    CAS  PubMed  Google Scholar 

  21. 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).

    CAS  PubMed  Google Scholar 

  22. 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).

    CAS  PubMed  Google Scholar 

  23. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Ng, L.G. et al. Migratory dermal dendritic cells act as rapid sensors of protozoan parasites. PLoS Pathog. 4, e1000222 (2008).

    PubMed  PubMed Central  Google Scholar 

  25. Springer, T.A. Adhesion receptors of the immune system. Nature 346, 425–434 (1990).

    CAS  PubMed  Google Scholar 

  26. Campbell, I.D. & Humphries, M.J. Integrin structure, activation, and interactions. Cold Spring Harb. Perspect. Biol. 3, 1–14 (2011).

    Google Scholar 

  27. 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).

    CAS  PubMed  Google Scholar 

  28. Ruoslahti, E. RGD and other recognition sequences for integrins. Annu. Rev. Cell Dev. Biol. 12, 697–715 (1996).

    CAS  PubMed  Google Scholar 

  29. 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).

    CAS  PubMed  Google Scholar 

  30. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  31. 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).

    CAS  PubMed  Google Scholar 

  32. Klebe, R.J. Isolation of a collagen-dependent cell attachment factor. Nature 250, 248–251 (1974).

    CAS  PubMed  Google Scholar 

  33. 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).

    CAS  PubMed  Google Scholar 

  34. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  35. 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).

    CAS  PubMed  Google Scholar 

  36. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  38. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Renkawitz, J. et al. Adaptive force transmission in amoeboid cell migration. Nat. Cell Biol. 11, 1438–1443 (2009).

    CAS  PubMed  Google Scholar 

  40. Friedl, P. & Wolf, K. Plasticity of cell migration: a multiscale tuning model. J. Cell Biol. 188, 11–19 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Sandig, H. et al. Fibronectin is a TH1-specific molecule in human subjects. J Allergy Clin Immunol 124, 528–535 (2009).

    CAS  PubMed  Google Scholar 

  42. Shulman, Z. et al. Lymphocyte crawling and transendothelial migration require chemokine triggering of high-affinity LFA-1 integrin. Immunity 30, 384–396 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Rutkowski, J.M. & Swartz, M.A. A driving force for change: interstitial flow as a morphoregulator. Trends Cell Biol. 17, 44–50 (2007).

    CAS  PubMed  Google Scholar 

  45. 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).

    CAS  PubMed  Google Scholar 

  46. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  47. 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).

    PubMed  PubMed Central  Google Scholar 

  48. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  49. 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).

    PubMed  Google Scholar 

  50. Masuoka, M. et al. Periostin promotes chronic allergic inflammation in response to Th2 cytokines. J. Clin. Invest. 122, 2590–2600 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  51. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  52. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  53. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  54. van der Flier, A. et al. Endothelial α5 and αV integrins cooperate in remodeling of the vasculature during development. Development 137, 2439–2449 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. Wennerberg, K. et al. β1 integrin-dependent and -independent polymerization of fibronectin. J. Cell Biol. 132, 227–238 (1996).

    CAS  PubMed  Google Scholar 

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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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Authors and Affiliations

Authors

Contributions

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.

Corresponding author

Correspondence to Deborah J Fowell.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

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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