Expression of stress proteins, adhesion molecules, and interleukin-8 in endothelial cells after preservation and reoxygenation.
Endothelial activation is a central feature of preservation-induced allograft injury. The present study aims at a quantitative assessment of stress proteins, adhesion molecules, and interleukin-8 in a cell culture-based model of organ preservation. Human umbilical vein endothelial cells were exposed to cold, hypoxic storage in University of Wisconsin (UW), histidine-tryptophane-ketoglutarate (HTK), and EuroCollins solutions for 8 h with subsequent rewarming/reoxygenation (rew/reox) for 1 and 4 h. A cell-based ELISA was designed for detection of heat shock proteins (HSP) 60 and 70, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and endothelial leukocyte adhesion molecule-1 (ELAM-1). Immunohistochemical staining was performed for comparison. Interleukin-8 was quantified by ELISA. HSP 70 was expressed after cold storage in HTK and EuroCollins solution and after rew/reox in all groups. A constitutive expression of HSP 60 was observed with further upregulation after rew/reox following cold storage in all experimental groups. ICAM-1 was clearly upregulated, but VCAM-1 showed only weak expression after cold storage and rew/reox. ELAM-1 was detectable in minimal amounts after cold storage but was considerably upregulated after 4 h of rew/reox. A significant increase of interleukin-8 release could be found after 4 h of rew/reox following storage in EuroCollins solution. Expression of stress proteins can be considered as a new parameter of preservation-associated endothelial activation. Apart from possible protective effects, allograft vasculopathy could be in part a consequence of the antigeneic potential of heat shock proteins connected with effects caused by adhesion molecules and inflammatory cytokines.
Eberl
T
T
Department of Transplant Surgery, D. Swarovski Research Laboratory, University Hospital Innsbruck, Anichstrasse 35, Innsbruck, A-6020, Austria.
Amberger
A
A
Herold
M
M
Hengster
P
P
Steurer
W
W
Hochleitner
B W
BW
Gnaiger
E
E
Margreiter
R
R
D000241:Adenosine; D000493:Allopurinol; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000378:metabolism; D003080:Cold Temperature; D019040:E-Selectin / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D005069:Evaluation Studies as Topic; D005978:Glutathione; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006639:Histidine; D006801:Humans; D006982:Hypertonic Solutions; D007328:Insulin; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism*; D007656:Ketoglutaric Acids; D019188:Organ Preservation Solutions; D010100:Oxygen; D011309:Preservation, Biological / Q000379:methods*; D011887:Raffinose; D014364:Tryptophan; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
D015815:Cell Adhesion Molecules; D018834:Chaperonin 60; D019040:E-Selectin; C037096:Euro-Collins' solution; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D006982:Hypertonic Solutions; D007328:Insulin; D016209:Interleukin-8; D007656:Ketoglutaric Acids; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D006639:Histidine; D000493:Allopurinol; D014364:Tryptophan; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D010100:Oxygen