title: Endothelial cell activation by sera containing HLA antibodies is mediated by interleukin-1.
abstract: BACKGROUND
  It has been suggested that antibodies which are associated with chronic pathological conditions such as chronic rejection and autoimmune diseases have the capacity to activate endothelial cells by induction and up-regulation of adhesion molecules. It has also been suggested that HLA antibodies formed by patients awaiting transplantation can activate endothelial cells. These antibodies include HLA and those that bind to endothelial cells.
  
  
  METHODS
  We have further investigated this phenomenon using monoclonal antibodies against HLA class I determinants and sera from aortic valve graft recipients, containing strong HLA antibodies. The effect of 24-hr incubation of antibodies/serum with human umbilical vein endothelial cells (HUVECs) on adhesion molecule expression was measured by flow cytometry.
  
  
  RESULTS
  HLA monoclonal antibodies had no effect on ICAM-1 expression on HUVECs. Five of 31 (16%) patients' sera caused strong up-regulation of adhesion molecules (ICAM-1, vascular cell adhesion molecule-1, and E-selectin) but this did not correlate with HLA specificity, IgG, or IgM binding to HUVECs. The activity, found in whole serum and IgG-depleted fractions was inhibited by neutralizing antibodies against interleukin (IL)-1beta and tumor necrosis factor-alpha. Examination of patient sera for presence of IL-1beta demonstrated high levels of IL-1beta in all five sera (range, 30 -500 U/ml) as well as in samples from an additional three patients.
  
  
  CONCLUSIONS
  The ability to activate endothelial cells detected in our patient sera was caused by cytokines and not antibody. Our observation that addition of cytokines to sera before separation into large and low molecular weight fractions demonstrated retention of cytokines in both fractions may be a confounding issue when investigating endothelial cell activation by patients' sera.
authors.affiliation: National Heart and Lung Institute, Imperial College of School of Medicine, Harefield Hospital, Middlesex, United Kingdom.
authors.forename: J D
authors.initials: JD
authors.lastname: Smith
authors.forename: M H
authors.initials: MH
authors.lastname: Yacoub
authors.forename: M L
authors.initials: ML
authors.lastname: Rose
mesh_terms: D000906:Antibodies / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000961:Antilymphocyte Serum / Q000494:pharmacology; D001021:Aortic Valve; D001790:Blood Physiological Phenomena / Q000187:drug effects*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000097:blood; D016207:Cytokines / Q000276:immunology / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D006680:HLA Antigens / Q000276:immunology*; D006349:Heart Valve Diseases / Q000097:blood / Q000601:surgery; D006350:Heart Valve Prosthesis; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007375:Interleukin-1 / Q000097:blood / Q000494:pharmacology*; D009500:Neutralization Tests; D014471:Umbilical Veins
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; D015815:Cell Adhesion Molecules; D016207:Cytokines; D006680:HLA Antigens; D007375:Interleukin-1; D018799:Intercellular Adhesion Molecule-1