Fabian A Helfritz1, Denisa Bojkova2, Verena Wanders2, Nina Kuklinski2, Sandra Westhaus2, Charlotte von Horn1, Ursula Rauen3, Anja Gallinat1, Hideo A Baba4, Andreas Skyschally5, Sandra Swoboda1, Volker Kinast6, Eike Steinmann6,7, Gerd Heusch5, Thomas Minor1, Philip Meuleman8, Andreas Paul1, Sandra Ciesek2,9. 1. General, Visceral and Transplantation Surgery, University Hospital Essen, University Duisburg-Essen, Germany. 2. Institute of Virology, University Hospital Essen, University Duisburg-Essen, Germany. 3. Institute of Physiological Chemistry, University Hospital Essen, University Duisburg-Essen, Germany. 4. Institute of Pathology, University Hospital Essen, University Duisburg-Essen, Germany. 5. Institute of Pathophysiology, West German Heart and Vascular Center, University Hospital Essen, University Duisburg-Essen, Germany. 6. Institute of Experimental Virology, Twincore, Centre for Experimental and Clinical Infection Research, Hannover, Germany. 7. Department of Molecular and Medical Virology, Ruhr-University Bochum, Germany. 8. Department of Clinical Chemistry, Microbiology and Immunology, Faculty of Medicine and Health Sciences, Ghent University, Belgium. 9. German Center for Infection Research (DZIF), External Partner Site Essen, Germany.
Abstract
Background: Although organ shortage is a rising problem, organs from hepatitis C virus (HCV) ribonucleic acid (RNA)-positive donors are not routinely transplanted in HCV-negative individuals. Because HCV only infects hepatocytes, other organs such as kidneys are merely contaminated with HCV via the blood. In this study, we established a protocol to reduce HCV virions during the cold ischemic time. Methods: Standard virological assays were used to investigate the effect of antivirals, including methylene blue (MB), in different preservation solutions. Kidneys from mini pigs were contaminated with Jc1 or HCV RNA-positive human serum. Afterwards, organs were flushed with MB. Hypothermic machine perfusion was used to optimize reduction of HCV. Results: Three different antivirals were investigated for their ability to inactivate HCV in vitro. Only MB completely inactivated HCV in the presence of all perfusion solutions. Hepatitis C virus-contaminated kidneys from mini pigs were treated with MB and hypothermic machine perfusion without any negative effect on the graft. Human liver-uPA-SCID mice did not establish HCV infection after inoculation with flow through from these kidneys. Conclusions: This proof-of-concept study is a first step to reduce transmission of infectious HCV particles in the transplant setting and might serve as a model for other relevant pathogens.
Background: Although organ shortage is a rising problem, organs from hepatitis C virus (HCV) ribonucleic acid (RNA)-positive donors are not routinely transplanted in HCV-negative individuals. Because HCV only infects hepatocytes, other organs such as kidneys are merely contaminated with HCV via the blood. In this study, we established a protocol to reduce HCV virions during the cold ischemic time. Methods: Standard virological assays were used to investigate the effect of antivirals, including methylene blue (MB), in different preservation solutions. Kidneys from mini pigs were contaminated with Jc1 or HCV RNA-positive human serum. Afterwards, organs were flushed with MB. Hypothermic machine perfusion was used to optimize reduction of HCV. Results: Three different antivirals were investigated for their ability to inactivate HCV in vitro. Only MB completely inactivated HCV in the presence of all perfusion solutions. Hepatitis C virus-contaminated kidneys from mini pigs were treated with MB and hypothermic machine perfusion without any negative effect on the graft. Human liver-uPA-SCID mice did not establish HCV infection after inoculation with flow through from these kidneys. Conclusions: This proof-of-concept study is a first step to reduce transmission of infectious HCV particles in the transplant setting and might serve as a model for other relevant pathogens.
Authors: Mathieu Gendrot; Julien Andreani; Isabelle Duflot; Manon Boxberger; Marion Le Bideau; Joel Mosnier; Priscilla Jardot; Isabelle Fonta; Clara Rolland; Hervé Bogreau; Sébastien Hutter; Bernard La Scola; Bruno Pradines Journal: Int J Antimicrob Agents Date: 2020-10-16 Impact factor: 15.441