title: Transfection of short-interfering RNA silences adhesion molecule expression on cardiac microvascular cells.
abstract: BACKGROUND
  Acute rejection reactions and the development of graft arterial disease are serious limitations after transplantation. Both are connected to the expression of adhesion molecules on the activated microvascular endothelium of the allograft.
  
  
  METHODS
  siRNA-mediated silencing of ESELE, ICAM-1, and VCAM-1 on human cardiac microvascular cells (HCMEC) was investigated in order to inhibit leukocyte-endothelial interactions. HCMEC were investigated for the time-dependent expression of ESELE, ICAM-1, and VCAM-1 after TNF-α stimulation and for siRNA mediated suppression using a nonviral transfecting approach. Furthermore, the effects of siRNA transfection on leukocyte binding to the endothelium were analyzed.
  
  
  RESULTS
  Transfection with siRNA induced a significant suppression of adhesion molecule expression, regardless of whether there had been a prior single or cocktail transfection of the sequences ( P < 0.05). The quantity of attaching leukocytes was significantly reduced after an equal silencing of adhesion molecules ( P < 0.05).
  
  
  CONCLUSIONS
  This investigation demonstrates that liposomal transfection of HCMEC with specific siRNA sequences is capable of both repressing adhesion molecule expression and of reducing subsequent leukocyte-endothelial actions.
authors.affiliation: Clinic for Thoracic, Cardiac and Vascular Surgery, University of Tübingen, Tübingen, Germany. tobias.walker@med.uni-tuebingen.de
authors.forename: T
authors.initials: T
authors.lastname: Walker
authors.forename: E
authors.initials: E
authors.lastname: Saup
authors.forename: A
authors.initials: A
authors.lastname: Nolte
authors.forename: P
authors.initials: P
authors.lastname: Simon
authors.forename: A
authors.initials: A
authors.lastname: Kornberger
authors.forename: V
authors.initials: V
authors.lastname: Steger
authors.forename: O
authors.initials: O
authors.lastname: Adolph
authors.forename: H-P
authors.initials: HP
authors.lastname: Wendel
mesh_terms: D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D003331:Coronary Vessels / Q000166:cytology / Q000378:metabolism*; D019040:E-Selectin / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008081:Liposomes; D055806:Microvessels / Q000166:cytology / Q000378:metabolism*; D009504:Neutrophils / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics
chemical_list: D015815:Cell Adhesion Molecules; D019040:E-Selectin; D008081:Liposomes; D034741:RNA, Small Interfering; C546936:SELE protein, human; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1