Elevated kallikrein activity in plasma from stable liver transplant recipients.
Extensive in vitro conversion of complement components C3 and C4 has been observed in EDTA plasma obtained from a number of stable orthotopic liver transplant recipients (LTR) [Clin. Chem. 45 (1999) 1190]. Consequently, we designed a chromogenic substrate (Ac-Ala-Gly-Leu-Thr-Arg-p-nitroanilide, AGLTR-pNA), based on the C1s cleavage site in complement component C4, in an attempt to identify the plasma proteinase(s) that cleaves C4 in vitro. Average peptidase activity in EDTA plasma obtained from stable LTR (n = 16) was significantly higher (P<0.01) than that in plasma from healthy non-transplant donors (n = 16). This peptidase activity was also detected using commercial substrates designed for specific coagulation proteinases. The plasma proteinase was not inhibited by hirudin, a thrombin inhibitor, but was inhibited by the plasma kallikrein inhibitor D-Phe-Phe-Arg-chloromethylketone, which fails to inhibit C1s. We concluded that the peptidase detected inLTR plasma, using chromogenic substrates including AGLTR-pNA, was plasma kallikrein. Western blot analysis confirmed the presence of kallikrein-alpha-2-macroglobulin complexes (alpha2M) in LTR plasmas. We also demonstrated that kallikrein was not the proteinase responsible for the in vitro cleavage of C4. Elevation of the plasma peptidase activity correlated significantly with recurrent hepatitis C virus (HCV) infection in these liver recipients with a P value <0.02. Significant correlation was not observed between complement activation (i.e. the C4a levels) and recurrent HCV infection (P>0.15); however, C4a levels did correlate with rejection (P<0.02). These results suggest that elevation in plasma peptidase activity and activation of complement do signal different pathological events in LTR, events that appear related to HCV-induced infection and immune tissue injury, respectively.
Hayashi
Joichiro
J
Division of Molecular Immunology, La Jolla Institute for Molecular Medicine, San Diego, CA 92121, USA.
Salomon
Daniel R
DR
Hugli
Tony E
TE
D000328:Adult; D002850:Chromatography, Gel; D003181:Complement C4 / Q000378:metabolism; D015934:Complement C4a / Q000378:metabolism; D005260:Female; D006084:Graft Rejection; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors / Q000097:blood*; D016031:Liver Transplantation / Q000009:adverse effects* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
D003181:Complement C4; D015842:Serine Proteinase Inhibitors; D015934:Complement C4a; D007610:Kallikreins