title: Effective suppression of class I major histocompatibility complex expression by the US11 or ICP47 genes can be limited by cell type or interferon-gamma exposure.
abstract: An impediment encountered in many viral-based gene therapy clinical trials has been the rapid destruction of the transgene by the host's immune response. The processing and presentation of antigens through the class I major histocompatibility complex (MHC) pathway is the initial specific response to viral infection. Disruption of the class I MHC pathway by herpes simplex virus (HSV) or the human cytomegalovirus (HCMV) results in a decrease of the CD8(+) cytotoxic T lymphocyte (CTL) response and prolongs survival of infected cells in the host. Two viral immune suppression genes that interfere with the class I MHC presentation pathway, the HSV type I ICP47 gene and HCMV US11 gene, were cloned and each incorporated into a retroviral vector. HSV ICP47 and HCMV US11 transgenes were expressed in multiple cells lines and compared for their abilities to reduce antigen presentation on the cell surface by class I MHC. Retroviral supernatants were used to transduce human, canine, and rat cell lines. Fluorescence-activated cell sorter (FACS) analysis of US11- and ICP47-transduced cell lines demonstrated substantial reductions in class I MHC cell surface expression in most cell lines except in rodent cells where ICP47 is nonfunctional. The decrease in the level of class I MHC expression for ICP47 transduced cell lines ranged from 31-98% relative to negative controls. US11 decreased class I cell surface MHC by 67-96%. When both ICP47 and US11 are expressed in human cells, a further reduction of class I MHC was observed. Next, human A375 melanoma cells were tested to determine if the resulting reduction in cell surface class I MHC would reduce in vitro cytotoxicity by CTL. A375 cells expressing either ICP47 or US11 demonstrated a twofold to threefold reduction of specific lysis by primed CD8(+) CTL. These data clearly establish an ability to convey immune protection to human cells by viral genes. However, further analysis demonstrated that interferon (IFN)-gamma could reverse part or all of the downregulation of class I MHC induced by the ICP47 or US11 genes. The ICP47 and US11 genes, when expressed in target cells, decrease class I MHC presentation and as such might be used in strategies to create local immunosuppression against transgenes or allografts.
authors.affiliation: The Iowa Cancer Research Foundation, Des Moines, IA 50309, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Radosevich
authors.forename: Tatiana
authors.initials: T
authors.lastname: Seregina
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Link
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival; D004285:Dogs; D015536:Down-Regulation; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D007165:Immunosuppression Therapy; D007371:Interferon-gamma / Q000494:pharmacology*; D008545:Melanoma / Q000235:genetics / Q000473:pathology; D016601:RNA-Binding Proteins / Q000235:genetics*; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D012878:Skin Neoplasms / Q000235:genetics / Q000473:pathology; D019076:Transgenes / Q000276:immunology; D014184:Transplantation, Homologous; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; D015395:Histocompatibility Antigens Class I; C105843:ICP47 protein, Herpes simplex virus; D017874:Immediate-Early Proteins; D016601:RNA-Binding Proteins; C074546:US11 protein, herpesvirus; D014764:Viral Proteins; D007371:Interferon-gamma