Porcine endogenous retrovirus released by a bioartificial liver infects primary human cells
Jan-Henning Frühauf,
Heike Mertsching,
Shibashish Giri,
Nils Roman Frühauf,
Augustinus Bader,
Jan-Henning Frühauf
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorHeike Mertsching
Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Medical School Hannover, Hannover, Germany
Search for more papers by this authorShibashish Giri
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorNils Roman Frühauf
German Foundation of Organ Transplantation, Hannover, Germany
Search for more papers by this authorAugustinus Bader
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorJan-Henning Frühauf,
Heike Mertsching,
Shibashish Giri,
Nils Roman Frühauf,
Augustinus Bader,
Jan-Henning Frühauf
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorHeike Mertsching
Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Medical School Hannover, Hannover, Germany
Search for more papers by this authorShibashish Giri
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorNils Roman Frühauf
German Foundation of Organ Transplantation, Hannover, Germany
Search for more papers by this authorAugustinus Bader
Department of Cell Techniques and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Leipzig, Germany
Search for more papers by this authorCorrespondence
Augustinus Bader, Full Professor, Cell Technologies and Applied Stem Cell Biology, Biomedical–Biotechnological Center (BBZ), Deutscher Platz 5, D-04103 Leipzig, Germany Tel: +49 341 97 31350, ext.: 31351Fax: +49 341 97 31359e-mail: [email protected]
Abstract
Background: Porcine endogenous retrovirus (PERV) remains a safety risk in pig-to-human xenotransplantation. There is no evidence of in vivo productive infection in humans because PERV is inactivated by human serum. However, PERV can infect human cell lines and human primary cells in vitro and inhibit human immune functions.
Aims: We investigated the potential of primary porcine liver cells to transmit PERV to primary human cells in a bioreactor-based bioartificial liver (BAL).
Methods: Primary human hepatocytes, endothelial cells and the human cell line HEK 293 were exposed to supernatants from BAL or from the porcine cell line PK-15. PERV polymerase-specific reverse-transcriptase polymerase chain reaction (RT-PCR) and PCR were used to investigate PERV transmission to human cells. An assay of RT activity was used to detect the presence of retrovirus in the supernatants of BAL, primary human hepatocytes and endothelial cells.
Results: Primary human hepatocytes (hHep), endothelial cells and HEK 293 cells were reproducibly infected by PERV, originating from primary porcine liver cells within the BAL and from PK-15 cells. Infected cells were positive for PERV-specific DNA and RNA after 8–10 days on an average, and RT activity was detectable in the supernatants of infected hHep and HEK 293 cells.
Conclusion: A risk of PERV infection in human cells is documented in this study, indicating that short-term contact of primary porcine liver cell supernatants with primary human cells could result in PERV transmission.
References
- 1 Patience C, Takeuchi Y, Weiss RA. Infection of human cells by an endogenous retrovirus of pigs. Nat Med 1997; 3: 282–6.
- 2 Kanai H, Marushima H, Kimura N, et al. Extracorporeal bioartificial liver using the radial-flow bioreactor in treatment of fatal experimental hepatic encephalopathy. Artif Organs 2006; 31: 148–51.
- 3 De Bartolo L, Bader A. Review of a flat membrane bioreactor as a bioartificial liver. Ann Transpl 2001; 6: 40–6.
- 4 Gerlach J, Schnoy N, Smith MD, et al. Hepatocyte culture between woven capillary networks: a microscopy study. Artif Organs 1994; 18: 226–30.
- 5 Flendrig LM, Sommeijer D, Ladiges NC, et al. Commercially available media for flushing extracorporeal bioartificial liver systems prior to connection to the patient's circulation: an in vitro comparative study in two and three dimensional porcine hepatocyte cultures. Int J Artif Organs 1998; 21: 467–72.
- 6 McKane BW, Ramachandran S, Yang J, et al. Xenoreactive anti-Gal alpha(1,3)Gal antibodies prevent porcine endogenous retrovirus infection of human in vivo. Hum Immunol 2003; 64: 708–17.
- 7 Rozga J, Podesta L, LePage E, et al. Control of cerebral oedema by total hepatectomy and extracorporeal liver support in fulminant hepatic failure. Lancet 1993; 342: 898–9.
- 8 Sussman NL, Chong MG, Koussayer T, et al. Reversal of fulminant hepatic failure using an extracorporeal liver assist device. Hepatology 1992; 16: 60–5.
- 9 Uchino J, Matsue H, Takahashi M, et al. A hybrid artificial liver system. Function of cultured monolayer pig hepatocytes in plasma from hepatic failure patients. ASAIO Trans 1991; 37: M337–8.
- 10 Kuddus R, Patzer JF II, Lopez R, et al. Clinical and laboratory evaluation of the safety of a bioartificial liver assist device for potential transmission of porcine endogenous retrovirus. Transplantation 2002; 73: 420–9.
- 11 Watanabe A, Sakai T, Sato S, et al. Clinical efficacy of lactulose in cirrhotic patients with and without subclinical hepatic encephalopathy. Hepatology 1997; 26: 1410–4.
- 12 Maki T, Monaco AP. Porcine islets xenotransplantation utilizing a vascularized bioartificial pancreas. Ann Transpl 1997; 2: 69–71.
- 13 Dinsmore JH, Manhart C, Raineri R, et al. No evidence for infection of human cells with porcine endogenous retrovirus (PERV) after exposure to porcine fetal neuronal cells. Transplantation 2000; 15: 1382–9.
- 14 Louz D, Bergmans HE, Loos BP, Hoeben RC. Reappraisal of biosafety risks posed by PERVs in xenotransplantation. Rev Med Virol 2008; 18: 53–65.
- 15 Weiss RA. Xenografts and retroviruses. Science 1999; 285: 1221–2.
- 16 Akiyoshi DE, Denaro M, Zhu H, et al. Identification of a full-length cDNA for an endogenous retrovirus of miniature swine. J Virol 1998; 72: 4503–7.
- 17 Van der Laan LJ, Lockey C, Griffeth BC, et al. Infection by porcine endogenous retrovirus after islet xenotransplantation in SCID mice. Nature 2000; 7: 90–4.
- 18 Martin U, Kiessig V, Blusch JH, et al. Expression of pig endogenous retrovirus by primary porcine endothelial cells and infection of human cells. Lancet 1998; 29: 692–4.
- 19 Wilson CA, Wong S, Muller J, et al. Type C retrovirus released from porcine primary peripheral blood mononuclear cells infects human cells. J Virol 1998; 72: 3082–7.
- 20 Paradis K, Langford G, Long Z, et al. Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. Science 1999; 285: 236–41.
- 21 Heneine W, Tibell A, Switzer WM, et al. No evidence of infection with porcine endogenous retrovirus in recipients of porcine islet-cell xenografts. Lancet 1998; 352: 695–9.
- 22 Patience C, Patton GS, Takeuchi Y, et al. No evidence of pig DNA or retroviral infection in patients with short-term extracorporeal connection to pig kidneys. Lancet 1998; 352: 699–701.
- 23 Fiebig U, Stephan O, Kurth R, Denner J. Neutralizing antibodies against conserved domains of p15E of porcine endogenous retroviruses: basis for a vaccine for xenotransplantation? Virology 2003; 307: 406–13.
- 24 Takeuchi Y, Patience C, Magre S, et al. Host range and interference studies of three classes of pig endogenous retrovirus. J Virol 1998; 72: 9986–91.
- 25 Ericsson TA, Takeuchi Y, Templin C, et al. Identification of receptors for pig endogenous retrovirus. Proc Natl Acad Sci USA 2003; 100: 6759–64.
- 26 Marcucci KT, Martina Y, Harrison F, Wilson CA, Salomon DR. Functional hierarchy of two L domains in porcine endogenous retrovirus (PERV) that influence release and infectivity. Virology 2008; 375: 637–45.
- 27 Le Tissier P, Stoye JP, Takeuchi Y, Patience C, Weiss RA. Two sets of human-tropic pig retrovirus. Nature 1997; 389: 681–2.
- 28 Blusch JH, Patience C, Martin U. Pig endogenous retroviruses and xenotransplantation. Xenotransplantation 2002; 9: 242–51.
- 29 Pitkin Z, Mullon C. Evidence of absence of porcine endogenous retrovirus (PERV) infection in patients treated with a bioartificial liver support system. Artif Organs 1999; 23: 829–33.
- 30 Falasca E, Adami V, Astori G, et al. Porcine endogenous retrovirus does not infect human cells using a bioartificial liver model system. Transpl Proc 2001; 33: 1780–1.
- 31 Kuddus R, Patzer JF, Lopez R, et al. Valuation of transmission of porcine endogenous retrovirus into patients subjected to hemoperfusion using an extracorporeal bioartificial liver support system. Transpl Proc 2001; 33: 1976.
- 32 Di Nicuolo G, Van De Kerkhove M-P, Hoekstra R, et al. No evidence of in vitro and in vivo porcine endogenous retrovirus infection after plasmapheresis through the AMC-bioartificial liver. Xenotransplantation 2005; 12: 286–92.
- 33 Dunn JC, Yarmush ML, Koebe HG, Tompkins RG. Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich configuration. FASEB J 1989; 3: 174–7.
- 34 Bader A, Zech K, Crome O, et al. Use of organotypical cultures of primary hepatocytes to analyse drug biotransformation in man and animals. Xenobiotica 1994; 24: 623–33.
- 35 Bader A, Hansen T, Kirchner G, et al. Primary porcine enterocyte and hepatocyte cultures to study drug oxidation reactions. Br J Pharmacol 2000; 129: 331–42.
- 36 Bader A, Steinhoff G, Strobl K, et al. Engineering of human vascular aortic tissue based on a xenogeneic starter matrix. Transplantation 2000; 70: 7–14.
- 37 Silver J, Maudru T, Fujita K, Repaske R. An RT-PCR assay for the enzyme activity of reverse transcriptase capable of detecting single virions. Nucleic Acids Res 1993; 21: 3593–4.
- 38 Liu Q, Liu Z, Dalakas E. Prevalence of porcine endogenous retrovirus in Chinese pig breeds and in patients treated with a porcine liver cell-based bioreactor. World J Gastroenterol 2005; 11: 4727–30.
- 39 Van de Kerkhove MP, Di Florio E, Scuderi V, et al. Bridging a patient with acute liver failure to liver transplantation by the AMC-bioartificial liver. Cell Transpl 2003; 12: 563–8.
- 40 Wang HH, Wang YJ, Liu HL, Liu J. Detection of PERV by polymerase chain reaction and its safety in bioartificial liver support system. World J Gastroenterol 2006; 12: 1287–91.
- 41 Irgang M, Sauer IM, Karlas A, et al. Porcine endogenous retroviruses: no infection in patients treated with a bioreactor based on porcine liver cells. J Clin Virol 2003; 28: 141–54.
- 42 Kardassis D, Zeillinger K, Pascher A, et al. Clinical extracorporeal hybrid liver support – phase I study with primary porcine liver cells. Xenotransplantation 2003; 10: 460–9.
- 43 Nishitai R, Ikai I, Shiotani T, et al. Absence of PERV infection in baboons after transgenic porcine liver perfusion. J Surg Res 2005; 124: 45–51.
- 44 Levy MF, Argaw T, Wilson CA, et al. No evidence of PERV infection in healthcare workers exposed to transgenic porcine liver extracorporeal support. Xenotransplantation 2007; 14: 309–15.
- 45 Hermida-Prieto M, Domenech N, Moscoso I, et al. Lack of cross-species transmission of porcine endogenous retrovirus (PERV) to transplant recipients and abattoir workers in contact with pigs. Transplantation 2007; 84: 548–50.
- 46 Tacke SJ, Kurth R, Denner J. Porcine endogenous retroviruses inhibit human immune cell function: risk for xenotransplantation? Virology 2000; 268: 87–93.
- 47 Tacke S, Specke V, Stephen O, et al. Porcine endogenous retroviruses: diagnostic assays and evidence for immunosuppressive properties. Transpl Proc 2000; 32: 1166.
- 48 Jungman A, Tonjes RR. Retrotransposition: another obstacle for xenotransplantation. Transpl Proc 2008; 40: 596–7.
- 49 Leib-Mösch C, Brack-Werner R, Werner T, et al. Endogenous retroviral elements in human DNA. Cancer Res 1990; 50: 5636–42.
- 50 Denner J. Is porcine endogenous retrovirus (PERV) transmission still relevant. Transpl Proc 2008; 40: 587–9.
- 51 Ericsson TA, Takeuchi Y, Templin C, et al. Identification of receptors for pig endogenous retrovirus. PNAS 2003; 100: 6759–64.
- 52 Mackey L. Feline leukaemia virus and its clinical effects in cats. Vet Rec 1975; 96: 5–11.
- 53 Teich N, Wyke J, Mak T, Bernstein A, Hardy W. Pathogenesis of retrovirus-induced disease. In RA Weiss, N Teich, HE Varmus, JM Coffin, eds. RNA Tumor Viruses, 2nd edn. New York, NY: Cold Spring Harbor Laboratory Press, 1984; 901–3.
- 54 Denner J. Immunosuppression by retroviruses: implications for xenotransplantation. Ann N Y Acad Sci 1998; 862: 76–86.
- 55 Weiss RA. Cross-species infections. Curr Top Microbiol Immunol 2003; 278: 47–71.
- 56 Bader A, Knop E, Boker KH, et al. Tacrolimus (FK 506) biotransformation in primary rat hepatocytes depends on extracellular matrix geometry. Naunyn Schmiedebergs. Arch Pharmacol 1996; 353: 461–73.
- 57 Nyberg SL, Hibbs JR, Hardin JA, Germer JJ, Persing DH. Transfer of porcine endogenous retrovirus across hollow fiber membranes: significance to a bioartificial liver. Transplantation 1999; 67: 1251–5.
- 58 Xu H, Sharma A, Okabe J, et al. Serologic analysis of anti-porcine endogenous retroviruses immune responses in humans after ex vivo transgenic pig liver perfusion. ASAIO J 2003; 49: 407–16.
- 59 Claudia R, Ballauff A, Lange R, Erhard J. Heterotopic auxiliary liver transplantation in a 3 year old boy with acute liver failure and aplastic anemia. Transplantation 1997; 64: 658–60.
- 60 Strain AJ, Neuberger JM. A bioartificial liver state of art. Science 2002; 295: 1005–9.
- 61 Mertsching H, Karim N, Haverich A, Bader A. Investigations on biological safety and immunologic aspects of chimeric bioartificial vessels in xenotransplantation. Transpl Proc 2000; 32: 1165.
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