title: Intracellular Kaposi's sarcoma-associated herpesvirus load determines early loss of immune synapse components.
abstract: Lifelong infection is a hallmark of all herpesviruses, and their survival depends on countering host immune defenses. The human gammaherpesvirus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes an array of proteins that contribute to immune evasion, including modulator of immune recognition 2 (MIR2), an E3 ubiquitin ligase. Exogenously expressed MIR2 downregulates the surface expression of several immune synapse proteins, including major histocompatibility complex (MHC) class 1, ICAM-1 (CD54), and PECAM (CD31). Although immunofluorescence assays detect this lytic gene in only 1 to 5% of cells within infected cultures, we have found that de novo infection of naive cells leads to the downregulation of these immune synapse components in a major proportion of the population. Investigating the possibility that low levels of MIR2 are responsible for this downregulation in the context of viral infection, we found that MIR2 transduction recapitulated the patterns of surface downregulation following de novo infection and that both MIR2 promoter activation, MIR2 expression level, and immune synapse component downregulation were proportional to the concentration of KSHV added to the culture. Additionally, MIR2-specific small interfering RNA reversed the downregulation effects. Finally, using a sensitive, high-throughput assay to detect levels of the virus in individual cells, we also observed that downregulation of MHC class I and ICAM-1 correlated with intracellular viral load. Together, these results suggest that the effects of MIR2 are gene dosage dependent and that low levels of this viral protein contribute to the widespread downregulation of immune-modulating cell surface proteins during the initial stages of KSHV infection.
authors.affiliation: Myles H. Thaler Center for AIDS and Human Retrovirus Research, University of Virginia Health Sciences, Charlottesville, VA 22908, USA.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Adang
authors.forename: Costin
authors.initials: C
authors.lastname: Tomescu
authors.forename: Wai K
authors.initials: WK
authors.lastname: Law
authors.forename: Dean H
authors.initials: DH
authors.lastname: Kedes
mesh_terms: D002460:Cell Line; D015536:Down-Regulation; D018628:Gene Dosage; D005786:Gene Expression Regulation; D020868:Gene Silencing; D019288:Herpesvirus 8, Human / Q000276:immunology* / Q000502:physiology*; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering; D019562:Viral Load; D014764:Viral Proteins / Q000502:physiology*; D017874:Immediate-Early Proteins
chemical_list: D015395:Histocompatibility Antigens Class I; D018799:Intercellular Adhesion Molecule-1; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D034741:RNA, Small Interfering; D014764:Viral Proteins; C405867:K5 protein, Human herpesvirus 8; D017874:Immediate-Early Proteins