title: Adenovirus DNA polymerase is recognized by human CD8+ T cells.
abstract: Donor lymphocytes have potential as a treatment for adenovirus (Ad) disease in haematopoietic stem cell transplant (SCT) recipients, but better understanding of Ad-specific T-cell responses is required. Most healthy adults exhibit memory T-cell responses to hexon, a capsid protein synthesized late after infection. However, since the Ad E3-19k downregulates major histocompatibility complex (MHC) class I molecules, cytotoxic T cells (CTLs) targeted to early viral proteins may be more effective in eliminating Ad-infected cells in vivo. Here we show that Ad-specific CTLs recognize the early region 2 proteins DNA polymerase (Pol) and DNA-binding protein (DBP). Firstly, memory Ad-specific CD8(+) T cells were amplified from healthy donors by in vitro stimulation with Ad-infected dendritic cells and found to exhibit MHC-restricted cytotoxicity to targets expressing Pol and DBP. Secondly, gamma interferon responses to HLA A2-binding motif peptides from Pol and DBP were directly detected in peripheral blood mononuclear cells (PBMCs) from a recently infected normal donor. Peptide-specific CTLs generated to Pol and DBP epitopes were confirmed to exhibit HLA A2-restricted killing of targets expressing Pol or DBP. Lastly, Pol-epitope-specific T cells were detected at similar or higher frequencies than hexon and DBP in three of three SCT recipients recovering from invasive Ad disease. Pol epitopes were well conserved among different Ad serotypes. Therefore, Pol is a promising target for immunotherapy of Ad disease.
authors.affiliation: Department of Microbiology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Amita
authors.initials: A
authors.lastname: Joshi
authors.forename: Jie
authors.initials: J
authors.lastname: Tang
authors.forename: Melanie
authors.initials: M
authors.lastname: Kuzma
authors.forename: John
authors.initials: J
authors.lastname: Wagner
authors.forename: Bijoyesh
authors.initials: B
authors.lastname: Mookerjee
authors.forename: Joanne
authors.initials: J
authors.lastname: Filicko
authors.forename: Matthew
authors.initials: M
authors.lastname: Carabasi
authors.forename: Neal
authors.initials: N
authors.lastname: Flomenberg
authors.forename: Phyllis
authors.initials: P
authors.lastname: Flomenberg
mesh_terms: D000256:Adenoviridae / Q000276:immunology*; D000328:Adult; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic; D004268:DNA-Binding Proteins / Q000276:immunology; D004259:DNA-Directed DNA Polymerase / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D016176:T-Lymphocyte Subsets / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D004268:DNA-Binding Proteins; D017361:Viral Nonstructural Proteins; D007371:Interferon-gamma; D004259:DNA-Directed DNA Polymerase