Effects of everolimus and HLA-G on cellular proliferation and neutrophil adhesion in an in vitro model of cardiac allograft vasculopathy

Am J Transplant. 2018 Dec;18(12):3038-3044. doi: 10.1111/ajt.15015. Epub 2018 Aug 13.

Abstract

Human leukocyte antigen-G (HLA-G) expression is modulated by immunosuppressant use and is associated with lower incidence of graft rejection and cardiac allograft vasculopathy (CAV). We examined whether everolimus induces HLA-G expression and inhibits human coronary artery smooth muscle cell (HCASMC) proliferation, a critical event in CAV. Also, we examined whether TNFα-stimulated neutrophil adhesion is inhibited by HLA-G on human coronary artery endothelial cells (HCAECs). HLA-G expression in HCASMCs following everolimus treatment was determined by western-blot densitometric analysis. HCASMCs proliferation following incubation with recombinant HLA-G was determined by automated cell counter detecting 2-10 µm particles. Assessment of recombinant HLA-G on neutrophil adhesion to HCAECs in response to TNF-α induced-injury was determined by nonstatic adhesion assays. HLA-G expression was upregulated in HCASMCs following everolimus exposure (1000 ng/ml; P < .05). HLA-G (500, 1000 ng/ml; both P < .05) reduced HCASMC proliferation and inhibited TNFα-stimulated neutrophil adhesion to endothelial cells at all concentrations (0.1-1 ng/ml; all P < .001). Our study reveals novel regulation of HLA-G by everolimus, by demonstrating HLA-G upregulation and subsequent inhibition of HCASMC proliferation. HLA-G is a potent inhibitor of neutrophil adhesion to HCAECs. Findings support HLA-G's importance and potential use in heart transplantation for preventative therapy or as a marker to identify patients at high risk for developing CAV.

Keywords: basic (laboratory) research/science; biomarker; clinical research/practice; coronary artery disease; heart (allograft) function/dysfunction; heart transplantation/cardiology; histocompatibility; immunosuppressant; immunosuppression/immune modulation; major histocompatibility complex (MHC).

MeSH terms

  • Allografts
  • Cell Adhesion*
  • Cell Proliferation / drug effects*
  • Cell Proliferation / physiology
  • Cells, Cultured
  • Coronary Vessels / drug effects
  • Coronary Vessels / immunology
  • Coronary Vessels / metabolism
  • Coronary Vessels / pathology*
  • Everolimus / pharmacology*
  • HLA-G Antigens / administration & dosage
  • HLA-G Antigens / immunology*
  • Humans
  • Immunosuppressive Agents / pharmacology
  • Models, Biological
  • Myocytes, Smooth Muscle / drug effects
  • Myocytes, Smooth Muscle / immunology
  • Myocytes, Smooth Muscle / metabolism
  • Myocytes, Smooth Muscle / pathology*
  • Neutrophils / drug effects
  • Neutrophils / immunology*
  • Vascular Diseases / drug therapy
  • Vascular Diseases / immunology
  • Vascular Diseases / metabolism
  • Vascular Diseases / pathology

Substances

  • HLA-G Antigens
  • Immunosuppressive Agents
  • Everolimus