title: Blockade of type I interferon (IFN) production by retroviral replicating vectors and reduced tumor cell responses to IFN likely contribute to tumor selectivity.
abstract: We developed a Moloney mouse leukemia virus (MLV)-based retroviral replicating vector (RRV), Toca 511, which has displayed tumor specificity in resected brain tumor material and blood in clinical trials. Here, we investigated the interaction between Toca 511 and human host cells, and we show that RRVs do not induce type I interferon (IFN) responses in cultured human tumor cells or cultured human primary cells. However, exogenous type I IFN inhibited RRV replication in tumor cells and induced IFN-regulated genes, albeit at a lower level than in primary cells. Unexpectedly, RRVs did not induce IFN-α production upon incubation in vitro with human plasmacytoid dendritic cells (pDCs), whereas lentivirus vector and heat-treated RRVs did. Coincubation of RRVs with heat-treated RRVs or with lentivirus vector suppressed IFN-α production in pDCs, suggesting that native RRV has a dominant inhibitory effect on type I IFN induction. This effect is sensitive to trypsin treatment. In addition, heat treatment inactivated that activity but exposed an immune-stimulatory activity. The immune-stimulating component is sensitive to deglycosidases, trypsin, and phospholipase C treatment. Experiments with retroviral nonreplicating vectors and virus-like particles demonstrated that the immunosuppressive activity is not associated with the amphotropic envelope or the glyco-Gag protein. In summary, our data provide evidence that RRVs do not directly trigger type I IFN responses in IFN-responsive tumor cells. Moreover, RRVs appear to carry a heat-labile component that actively suppresses activation of cellular innate immune responses in pDCs. Inhibition of IFN induction by RRVs and the reduced response to IFN should facilitate tumor-specific infection in vivo.
  
  
  OBJECTIVE
  RRVs have a convincing preference for replicating in tumor cells in animal models, and we observed similar preferences in the initial treatment of human glioblastoma patients. This study investigates the basis for the interaction between RRV and human host cells (tumor versus nontumor) in vitro. We found that RRVs do not trigger an IFN-α/β response in tumor cells, but the cells are capable of responding to type I IFNs and of producing them when stimulated with known agonists. Surprisingly, the data show that RRVs can actively inhibit induction of cellular innate immunity and that this inhibitory activity is heat labile and trypsin sensitive and not attributable to the envelope protein. These data partially explain the observed in vivo tumor specificity.
authors: Tocagen Inc., San Diego, California, USA.
authors: Amy H
authors: AH
authors: Lin
authors: Tocagen Inc., San Diego, California, USA.
authors: Cindy
authors: C
authors: Burrascano
authors: Tocagen Inc., San Diego, California, USA.
authors: Par L
authors: PL
authors: Pettersson
authors: Tocagen Inc., San Diego, California, USA.
authors: Carlos E
authors: CE
authors: Ibañez
authors: Tocagen Inc., San Diego, California, USA.
authors: Harry E
authors: HE
authors: Gruber
authors: Tocagen Inc., San Diego, California, USA djolly@tocagen.com.
authors: Douglas J
authors: DJ
authors: Jolly
mesh_terms: D002478:Cells, Cultured; D006801:Humans; D007370:Interferon Type I / Q000378:metabolism*; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000276:immunology* / Q000502:physiology*; D009369:Neoplasms / Q000276:immunology*; D014779:Virus Replication
chemical_list: D007370:Interferon Type I