title: Clinical and laboratory evaluation of the safety of a bioartificial liver assist device for potential transmission of porcine endogenous retrovirus.
abstract: BACKGROUND
  The potential risk of transmission of porcine endogenous retroviruses (PERV) from xenogeneic donors into humans has been widely debated. Because we were involved in a phase I/II clinical trial using a bioartificial liver support system (BLSS), we proceeded to evaluate the biosafety of this device.
  
  
  METHODS
  The system being evaluated contains primary porcine hepatocytes freshly isolated from pathogen-free, purpose-raised herd. Isolated hepatocytes were installed in the shell, which is separated by a semipermeable membrane (100-kD nominal cutoff) from the lumen through which the patients' whole blood is circulated. Both before and at defined intervals posthemoperfusion, patients' blood was obtained for screening. Additionally, effluent collected from a clinical bioreactor was analyzed. The presence of viral particles was estimated by reverse transcriptase-polymerase chain reaction (RT-PCR) and RT assays. For the detection of pig genomic and mitochondrial DNA, sequence-specific PCR (SS-PCR) was used. Finally, the presence of infectious viral particles in the samples was ascertained by exposure to the PERV-susceptible human cell line HEK-293.
  
  
  RESULTS
  PERV transcripts, RT activity, and infectious PERV particles were not detected in the luminal effluent of a bioreactor. Culture supernatant from untreated control or mitogen-treated porcine hepatocytes (cleared of cellular debris) also failed to infect HEK-293 cell lines. Finally, RT-PCR, SS-PCR, and PERV-specific RT assay detected no PERV infection in the blood samples obtained from five study patients both before and at various times post-hemoperfusion.
  
  
  CONCLUSIONS
  Although longer patient follow-up is required and mandated to unequivocally establish the biosafety of this device and related bioartificial organ systems, these analyses support the conclusion that when used under standard operational conditions, the BLSS is safe.
authors.affiliation: Thomas E. Starzl Transplantation Institute, and the Department of Surgery, University of Pittsburgh Medical Center-Health System, Pittsburgh, Pennsylvania, USA.
authors.forename: R
authors.initials: R
authors.lastname: Kuddus
authors.forename: J F
authors.initials: JF
authors.lastname: Patzer
authors.forename: R
authors.initials: R
authors.lastname: Lopez
authors.forename: G V
authors.initials: GV
authors.lastname: Mazariegos
authors.forename: B
authors.initials: B
authors.lastname: Meighen
authors.forename: D J
authors.initials: DJ
authors.lastname: Kramer
authors.forename: A S
authors.initials: AS
authors.lastname: Rao
mesh_terms: D000818:Animals; D019149:Bioreactors; D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis; D020077:Endogenous Retroviruses / Q000302:isolation & purification*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D019164:Liver, Artificial / Q000009:adverse effects*; D012367:RNA, Viral / Q000032:analysis; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012449:Safety; D013552:Swine / Q000821:virology*; D014771:Virion / Q000302:isolation & purification
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase