The integrin-linked kinase is required for chemokine-triggered high-affinity conformation of the neutrophil β2-integrin LFA-1

Blood. 2020 Nov 5;136(19):2200-2205. doi: 10.1182/blood.2020004948.

Abstract

Neutrophil adhesion and extravasation into tissue at sites of injury or infection depend on binding of the integrin lymphocyte function-associated antigen 1 (LFA-1) to ICAM-1 expressed on activated endothelial cells. The activation-dependent conformational change of LFA-1 to the high-affinity conformation (H+) requires kindlin-3 binding to the β2-integrin cytoplasmic domain. Here we show that genetic deletion of the known kindlin interactor integrin-linked kinase (ILK) impaired neutrophil adhesion and extravasation in the cremaster muscle and in a clinically relevant model of renal ischemia reperfusion injury. Using in vitro microfluidic adhesion chambers and conformation-specific antibodies, we show that knockdown of ILK in HL-60 cells reduced the conformational change of β2-integrins to the H+ conformation. Mechanistically, we found that ILK was required for protein kinase C (PKC) membrane targeting and chemokine-induced upregulation of its kinase activity. Moreover, PKC-α deficiency also resulted in impaired leukocyte adhesion in bone marrow chimeric mice. Mass spectrometric and western blot analyses revealed stimulation- and ILK-dependent phosphorylation of kindlin-3 upon activation. In summary, our data indicate an important role of ILK in kindlin-3-dependent conformational activation of LFA-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Kidney Injury / drug therapy
  • Acute Kidney Injury / etiology
  • Acute Kidney Injury / immunology
  • Acute Kidney Injury / metabolism*
  • Animals
  • CD18 Antigens / chemistry
  • CD18 Antigens / metabolism*
  • Cell Adhesion
  • Chemokines / pharmacology*
  • Disease Models, Animal
  • HL-60 Cells
  • Humans
  • Leukocytes / drug effects
  • Leukocytes / immunology
  • Leukocytes / metabolism
  • Lymphocyte Function-Associated Antigen-1 / chemistry
  • Lymphocyte Function-Associated Antigen-1 / metabolism*
  • Membrane Proteins / metabolism*
  • Mice
  • Neoplasm Proteins / metabolism*
  • Neutrophils / drug effects
  • Neutrophils / immunology
  • Neutrophils / metabolism
  • Phosphorylation
  • Protein Kinase C / metabolism*
  • Protein-Serine-Threonine Kinases / metabolism*
  • Reperfusion Injury / complications
  • Signal Transduction

Substances

  • CD18 Antigens
  • Chemokines
  • FERMT3 protein, human
  • Lymphocyte Function-Associated Antigen-1
  • Membrane Proteins
  • Neoplasm Proteins
  • integrin-linked kinase
  • Protein-Serine-Threonine Kinases
  • Protein Kinase C