Very Late Antigen Integrin Activity on T Cells

Summary

Principal Investigator: Yoji Shimizu
Abstract: Integrin adhesion receptors play a vital role in cell function and development by mediating the adhesion of cells to other cells and to extracellular matrix proteins. In the immune system, the functional activity of integrins expressed on T lymphocytes is dynamically regulated by the activation state of the T cell in order to promote transient periods of adhesion that facilitate T cell trafficking and antigen recognition in tissue. Stimulation of the antigen-specific CD3/T cell receptor (TCP) complex initiates a signaling cascade that results in a rapid increase in the functional activity of beta l and beta 2 integrins without increases in integrin levels on the cell surface. TCR-mediated integrin activation can be mimicked by phorbol esters such as PMA, implicating effectors downstream of protein kinase C (PKC) in this process. During the prior funding period, we identified a novel adapter protein function for the PKC effector protein kinase D1 (PKD1) in PMA- and TCR-mediated enhancement of beta l integrin function that is associated with increased beta l integrin clustering and activation of the small GTPase Rap1. We propose that the carboxy-terminal end of the beta l integrin cytoplasmic domain regulates integrin function in response to T cell activation by nucleating the formation of a membrane-localized complex consisting of PKD1, Rap1 and the Rap1 guanine nucleotide exchange factor C3G that controls Rap1 activation and subsequent integrin clustering. We propose the use of novel in vitro cell systems and genetically modified mice in order to test this hypothesis and its relevance to T cell function during immune responses in intact animals. In Aim 1, we will define the structural basis for interactions between the beta l integrin cytoplasmic domain and the PKD1/Rap1/C3G complex and determine if the beta l integrin tail is sufficient to control Rap1 activity. In Aim 2, we will use conditional gene targeting and adoptive transfer approaches to define the function of beta l integrin expression in controlling integrin- dependent responses of T cells to antigen challenge in mice. We will specifically determine if beta2 integrin function is dependent on expression of a beta l integrin subunit that couples effectively to PKD1 and Rap1. In Aim 3, we will use RNA interference and adenoviral-mediated gene transfer techniques to elucidate the function of PKD1 in regulating integrin function in vitro and T cell activation responses to antigen in vivo. Together, these studies will define a novel function for both PKD1 and the beta l integrin cytoplasmic domain in regulating integrin function in T cells and will enhance our ability to develop novel therapies that can specifically modulate T cell immune responses.
Funding Period: ----------------2010 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. pmc The WAVE2 complex regulates actin cytoskeletal reorganization and CRAC-mediated calcium entry during T cell activation
    Jeffrey C Nolz
    Department of Immunology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA
    Curr Biol 16:24-34. 2006
  2. pmc Formins regulate the actin-related protein 2/3 complex-independent polarization of the centrosome to the immunological synapse
    Timothy S Gomez
    Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA
    Immunity 26:177-90. 2007
  3. pmc WAVE2 regulates high-affinity integrin binding by recruiting vinculin and talin to the immunological synapse
    Jeffrey C Nolz
    Department of Immunology and Division of Oncology Research, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA
    Mol Cell Biol 27:5986-6000. 2007
  4. ncbi T-cell receptor signaling to integrins
    Brandon J Burbach
    Department of Laboratory Medicine and Pathology, Center for Immunology, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    Immunol Rev 218:65-81. 2007
  5. ncbi Adhesion and degranulation-promoting adapter protein (ADAP) positively regulates T cell sensitivity to antigen and T cell survival
    Kristen L Mueller
    Department of Laboratory Medicine and Pathology, Center for Immunology, Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 179:3559-69. 2007
  6. pmc Distinct regulation of integrin-dependent T cell conjugate formation and NF-kappa B activation by the adapter protein ADAP
    Brandon J Burbach
    Department of Laboratory Medicine and Pathology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 181:4840-51. 2008
  7. pmc The WAVE2 complex regulates T cell receptor signaling to integrins via Abl- and CrkL-C3G-mediated activation of Rap1
    Jeffrey C Nolz
    Department of Immunology, College of Medicine, Mayo Clinic, Rochester, MN 55905, USA
    J Cell Biol 182:1231-44. 2008
  8. pmc Control of alpha4beta7 integrin expression and CD4 T cell homing by the beta1 integrin subunit
    Christopher C DeNucci
    Department of Laboratory Medicine and Pathology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 184:2458-67. 2010
  9. pmc NF-kappaB activation in T cells requires discrete control of IkappaB kinase alpha/beta (IKKalpha/beta) phosphorylation and IKKgamma ubiquitination by the ADAP adapter protein
    Rupa Srivastava
    Department of Laboratory Medicine and Pathology, Center for Immunology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA
    J Biol Chem 285:11100-5. 2010
  10. pmc Integrin function in T-cell homing to lymphoid and nonlymphoid sites: getting there and staying there
    Christopher C DeNucci
    Department of Laboratory Medicine and Pathology, Center for Immunology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN, USA
    Crit Rev Immunol 29:87-109. 2009

Detail Information

Publications11

  1. pmc The WAVE2 complex regulates actin cytoskeletal reorganization and CRAC-mediated calcium entry during T cell activation
    Jeffrey C Nolz
    Department of Immunology, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA
    Curr Biol 16:24-34. 2006
    ..However, whether these proteins participate in actin reorganization at the IS or signaling pathways in T cells has not been investigated...
  2. pmc Formins regulate the actin-related protein 2/3 complex-independent polarization of the centrosome to the immunological synapse
    Timothy S Gomez
    Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA
    Immunity 26:177-90. 2007
    ..Altogether, our results have identified Arp2/3 complex-independent cytoskeletal reorganization events in T lymphocytes and indicate that formins are essential cytoskeletal regulators of centrosome polarity in T cells...
  3. pmc WAVE2 regulates high-affinity integrin binding by recruiting vinculin and talin to the immunological synapse
    Jeffrey C Nolz
    Department of Immunology and Division of Oncology Research, Mayo Clinic College of Medicine, 200 First Street SW, Rochester, MN 55905, USA
    Mol Cell Biol 27:5986-6000. 2007
    ....
  4. ncbi T-cell receptor signaling to integrins
    Brandon J Burbach
    Department of Laboratory Medicine and Pathology, Center for Immunology, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    Immunol Rev 218:65-81. 2007
    ..These events coordinate changes in integrin conformation and clustering that result in enhanced integrin functional activity following TCR stimulation...
  5. ncbi Adhesion and degranulation-promoting adapter protein (ADAP) positively regulates T cell sensitivity to antigen and T cell survival
    Kristen L Mueller
    Department of Laboratory Medicine and Pathology, Center for Immunology, Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 179:3559-69. 2007
    ....
  6. pmc Distinct regulation of integrin-dependent T cell conjugate formation and NF-kappa B activation by the adapter protein ADAP
    Brandon J Burbach
    Department of Laboratory Medicine and Pathology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 181:4840-51. 2008
    ....
  7. pmc The WAVE2 complex regulates T cell receptor signaling to integrins via Abl- and CrkL-C3G-mediated activation of Rap1
    Jeffrey C Nolz
    Department of Immunology, College of Medicine, Mayo Clinic, Rochester, MN 55905, USA
    J Cell Biol 182:1231-44. 2008
    ..These findings identify a previously unknown mechanism by which the WAVE2 complex regulates TCR signaling to Rap1 and integrin activation...
  8. pmc Control of alpha4beta7 integrin expression and CD4 T cell homing by the beta1 integrin subunit
    Christopher C DeNucci
    Department of Laboratory Medicine and Pathology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 184:2458-67. 2010
    ..Thus, the preferential association of beta1 integrin with alpha4 integrin suppresses alpha4beta7 integrin expression and regulates the localization of memory CD4 T cells...
  9. pmc NF-kappaB activation in T cells requires discrete control of IkappaB kinase alpha/beta (IKKalpha/beta) phosphorylation and IKKgamma ubiquitination by the ADAP adapter protein
    Rupa Srivastava
    Department of Laboratory Medicine and Pathology, Center for Immunology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA
    J Biol Chem 285:11100-5. 2010
    ..In contrast, only the CARMA1 binding site in ADAP is required for ubiquitination of IKKgamma. Thus, distinct sites within ADAP control two key activation responses that are required for NF-kappaB activation in T cells...
  10. pmc Integrin function in T-cell homing to lymphoid and nonlymphoid sites: getting there and staying there
    Christopher C DeNucci
    Department of Laboratory Medicine and Pathology, Center for Immunology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN, USA
    Crit Rev Immunol 29:87-109. 2009
    ....
  11. pmc β1 integrin is critical for the maintenance of antigen-specific CD4 T cells in the bone marrow but not long-term immunological memory
    Christopher C DeNucci
    Department of Laboratory Medicine and Pathology, Center for Immunology, Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA
    J Immunol 186:4019-26. 2011
    ..Thus, β(1) integrin-dependent maintenance of memory CD4 T cells in the bone marrow is not required for long-term CD4 T cell memory...