title: No in vivo infection of triple immunosuppressed non-human primates after inoculation with high titers of porcine endogenous retroviruses.
abstract: Porcine endogenous retroviruses (PERVs) released from pig tissue can infect selected human cells in vitro and therefore represent a safety risk for xenotransplantation using pig cells, tissues, or organs. Although PERVs infect cells of numerous species in vitro, attempts to establish reliable animal models failed until now. Absence of PERV transmission has been shown in first experimental and clinical xenotransplantations; however, these trials suffered from the absence of long-term exposure (transplant survival) and profound immunosuppression.
  
  
  METHODS
  We conducted infectivity studies in rhesus monkeys, pig-tailed monkeys, and baboons under chronic immunosuppression with cyclosporine A, methylprednisolone, and the rapamycin derivative. These species were selected because they are close to the human species and PERVs can be transmitted in vitro to cells of these species. In addition, the animals received twice, a C1 esterase inhibitor to block complement activation before inoculation of PERV. In order to overcome the complications of microchimerism, animals were inoculated with high titers of cell-free PERV. In addition, to enable transmission via cell-cell contact, some animals also received virus-producing cells. For inoculation the primate cell-adapted strain PERV/5 degrees was used which is characterized by a high infectious titer. Produced on human cells, this virus does not express alpha 1,3 Gal epitopes, does not contain porcine antigens on the viral surface and is therefore less immunogenic in non-human primates compared with pig cell-derived virus. Finally, we present evidence that PERV/5 degrees productively infects cells from baboons and rhesus monkeys.
  
  
  RESULTS
  In a follow-up period of 11 months, no antibody production against PERV and no integration of proviral DNA in blood cells was observed. Furthermore, no PERV sequences were detected in the DNA of different organs taken after necropsy.
  
  
  CONCLUSIONS
  These results indicate that in a primate model, in the presence of chronic immunosuppression, neither the inoculation of cell-free nor cell-associated PERV using a virus already adapted to primate cells results in an infection; this is despite the fact that peripheral blood mononuclear cells of the same animals are infectible in vitro.
authors.affiliation: Robert Koch-Institute, Berlin, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Specke
authors.forename: Roland
authors.initials: R
authors.lastname: Plesker
authors.forename: James
authors.initials: J
authors.lastname: Wood
authors.forename: Cheick
authors.initials: C
authors.lastname: Coulibaly
authors.forename: Kristen
authors.initials: K
authors.lastname: Suling
authors.forename: Clive
authors.initials: C
authors.lastname: Patience
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kurth
authors.forename: Henk-Jan
authors.initials: HJ
authors.lastname: Schuurman
authors.forename: Joachim
authors.initials: J
authors.lastname: Denner
mesh_terms: D000818:Animals; D016572:Cyclosporine / Q000627:therapeutic use; D004279:DNA, Viral / Q000097:blood; D004359:Drug Therapy, Combination; D020077:Endogenous Retroviruses / Q000235:genetics / Q000472:pathogenicity*; D005260:Female; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D008253:Macaca mulatta / Q000276:immunology*; D008254:Macaca nemestrina / Q000276:immunology*; D008297:Male; D008775:Methylprednisolone / Q000627:therapeutic use; D010215:Papio / Q000276:immunology*; D018419:Primate Diseases / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D012192:Retroviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D020123:Sirolimus / Q000627:therapeutic use; D013552:Swine / Q000821:virology*; D014183:Transplantation, Heterologous
chemical_list: D004279:DNA, Viral; D007166:Immunosuppressive Agents; D016572:Cyclosporine; D020123:Sirolimus; D008775:Methylprednisolone