title: Elastase and cathepsin G from primed leukocytes cleave vascular endothelial cadherin in hemodialysis patients.
abstract: OBJECTIVE
  To test the hypothesis that primed PMNLs in blood of chronic kidney disease patients release the active form of elastase and cathepsin G causing degradation of vital proteins and promote tissue damage.
  
  
  METHODS
  RT-PCR, immunocytochemical staining, immunoblotting, and FACS analyses were used to study these enzymes in hemodialysis patients (HD) versus healthy normal controls (NC). Using PMNLs and endothelial cells cocultivation system we measure the effect of HD PMNLs on the endothelial VE-cadherin, an essential protein for maintaining endothelial integrity.
  
  
  RESULTS
  Levels of elastase and cathepsin G were reduced in PMNLs of HD patients, while mRNA enzymes levels were not different. Elevated levels of the active form of these enzymes were found in blood of HD patients compared to NC.HD plasma had higher levels of soluble VE-cadherin present in three molecular forms: whole 140 kDa molecule and two fragments of 100 and 40 kDa. Cocultivation studies showed that primed PMNLs cleave the endothelial cadherin, resulting in a 100 kDa fragment.
  
  
  CONCLUSIONS
  Elastase and cathepsin G are elevated in the plasma of HD patients, originating from primed PMNLs. In these patients, chronic elevation of these enzymes contributes to cleavage of VE-cadherin and possible disruption of endothelial integrity.
authors.affiliation: Bruce Rappaport School of Medicine, Technion, 31096 Haifa, Israel ; Eliachar Research Laboratory, Western Galilee Hospital, 22100 Naharyia, Israel.
authors.forename: Meital
authors.initials: M
authors.lastname: Cohen-Mazor
authors.affiliation: Bruce Rappaport School of Medicine, Technion, 31096 Haifa, Israel ; Eliachar Research Laboratory, Western Galilee Hospital, 22100 Naharyia, Israel.
authors.forename: Rafi
authors.initials: R
authors.lastname: Mazor
authors.affiliation: Nephrology Unit, Western Galilee Hospital, 22100 Nahariya, Israel ; Faculty of Medicine in the Galilee, Bar Ilan University, 13100 Safed, Israel.
authors.forename: Batya
authors.initials: B
authors.lastname: Kristal
authors.affiliation: Eliachar Research Laboratory, Western Galilee Hospital, 22100 Naharyia, Israel ; Faculty of Medicine in the Galilee, Bar Ilan University, 13100 Safed, Israel.
authors.forename: Shifra
authors.initials: S
authors.lastname: Sela
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism*; D056649:Cathepsin G / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000293:injuries / Q000378:metabolism* / Q000473:pathology; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D007962:Leukocytes / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D010196:Pancreatic Elastase / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D006435:Renal Dialysis; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D056649:Cathepsin G; D010196:Pancreatic Elastase; D012333:RNA, Messenger; D000099152:Cadherin 5; C534094:CTSG protein, human