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Hepatitis C‐induced hepatocyte apoptosis following liver transplantation is enhanced by immunosuppressive agents
Author(s) -
Lim E. J.,
Chin R.,
Nachbur U.,
Silke J.,
Jia Z.,
Angus P. W.,
Torresi J.
Publication year - 2016
Publication title -
journal of viral hepatitis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 100
eISSN - 1365-2893
pISSN - 1352-0504
DOI - 10.1111/jvh.12541
Subject(s) - sirolimus , tacrolimus , apoptosis , liver transplantation , hepatocyte , hepatitis c , viability assay , calcineurin , transplantation , programmed cell death , medicine , immunology , biology , cancer research , in vitro , biochemistry
Summary In recurrent hepatitis C ( HCV ) post‐liver transplantation ( OLT ), the combination of immunosuppressants and HCV is postulated to increase hepatocyte apoptosis and liver fibrosis. We evaluated hepatocyte apoptosis within the liver tissue of patients with post OLT HCV recurrence compared to HCV ‐negative individuals and correlated these findings with the effects of immunosuppressants on HCV ‐induced cell death and its inhibition in primary mouse hepatocytes ( PM oH). Liver biopsies from patients with and without HCV were evaluated by immunohistochemistry for markers of apoptosis M30 Cyto DEATH (M30) and cleaved PARP (cl PARP ). PM oH from C57 BL /6 mice were infected with recombinant adenoviruses ( rAdHCV ) that expressed HCV proteins in hepatocytes. Infected cells were treated with cyclosporine, tacrolimus, sirolimus and/or MMF with or without pan‐caspase inhibitor Q‐ VD ‐Oph. Cell viability and apoptosis were evaluated using crystal violet assays and Western immunoblots probed for cleaved caspase‐3 (clCas3) and cl PARP . Both M30 and cl PARP were increased in the liver biopsies of patients with post OLT HCV recurrence compared to HCV ‐negative individuals. Treatment of rAdHCV ‐infected PM oH with cyclosporine, tacrolimus or sirolimus reduced cell viability and increased clCas3 and cl PARP compared to rAdHCV infection alone. Addition of MMF to cyclosporine, tacrolimus or sirolimus further reduced cell viability and increased clCas3 and cl PARP . Q‐ VD ‐Oph improved cell viability in HCV ‐infected PM oH treated with immunosuppressants alone and in combination and reduced clCas3 and cl PARP by approximately 90%. Immunosuppressive agents, especially in combination, enhanced apoptosis in HCV ‐infected hepatocytes. The finding that Q‐ VD ‐Oph reversed hepatocyte death suggests that treatments utilizing apoptosis inhibition might reduce liver injury in post OLT HCV recurrence.