Quantitative temporal viromics: an approach to investigate host-pathogen interaction.
Bottom Line: QTV predicted natural killer and T cell ligands, as well as 29 viral proteins present at the cell surface, potential therapeutic targets.Temporal profiles of >80% of HCMV canonical genes and 14 noncanonical HCMV open reading frames were defined.QTV is a powerful method that can yield important insights into viral infection and is applicable to any virus with a robust in vitro model.
Affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA; Cambridge Institute for Medical Research, University of Cambridge, Hills Road, Cambridge CB2 0XY, UK. Electronic address: email@example.com.Show MeSH
Related in: MedlinePlus
Mentions: We confirmed temporal profiles of protocadherins PCDHγC3 and FAT1 in addition to eight other cell-surface proteins by flow cytometry and immunoblot (Figures 4D and S4A, S4C, and S4E). To provide proof-of-principle validation of our functional predictions, we performed siRNA-mediated knockdown of the C-type lectin CLEC1A (Table S5A), which is downregulated 7-fold by HCMV and is a potential NK ligand by homology to CLEC2D. Polyclonal NK-cells from three of three independent donors demonstrated reduced degranulation to autologous targets upon knockdown of CLEC1A suggesting that this molecule may be a novel activating NK ligand (Figure S4D). We observed similar results for the protocadherin FAT1, providing initial confirmatory evidence that members of this family may indeed be involved in immunoregulation (Figure S4E). CEACAM-1 (immunoglobulin family) was upregulated 20-fold by HCMV infection and may have roles in T cell regulation. Using a blocking antibody, we demonstrated increased degranulation of CD8+ T cells specific for an HCMV HLA-A2 restricted peptide epitope suggesting that upregulation of this molecule in infected cells may inhibit cytotoxic T cell-mediated lysis (Figure S4F).
Affiliation: Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA; Cambridge Institute for Medical Research, University of Cambridge, Hills Road, Cambridge CB2 0XY, UK. Electronic address: firstname.lastname@example.org.