title: Genome-wide screening of human T-cell epitopes in influenza A virus reveals a broad spectrum of CD4(+) T-cell responses to internal proteins, hemagglutinins, and neuraminidases.
abstract: We performed a genome-wide screening for T-cell epitopes using synthetic peptides that encompass all of the influenza A viral proteins, including subtype variants for hemagglutinin (HA; H1, H3, and H5) and neuraminidase (NA; human and avian N1 and N2) proteins, based on the sequence information of recently circulating strains. We identified a total of 83 peptides, 54 of them novel, to which specific T cells were detectable in interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot assays using peripheral blood mononuclear cells from four healthy adult donors. The surface glycoproteins, HA and NA, major components of vaccines, expressed many T-cell epitopes. HA and matrix protein 1 expressed more T-cell epitopes than other viral proteins, most of which were recognized by CD4(+) T cells. We established several cytotoxic CD4(+) T-cell lines from these donors. We also analyzed H1 and H3 HA-specific T-cell responses using the peripheral blood mononuclear cells of 30 hospital workers. Fifty-three percent of donors gave a positive response to H3 HA peptides, whereas 17% gave a positive response to H1 HA peptides. Our genome-wide screening is useful in identifying T-cell epitopes and is complementary to the approach based on the predicted binding peptides to well-studied HLA-A, -B, and -DR alleles.
authors.affiliation: Center for Infectious Disease and Vaccine Research, University of Massachusetts Medical School, Worcester, MA 01655, USA.
authors.forename: Jenny Aurielle B
authors.initials: JA
authors.lastname: Babon
authors.forename: John
authors.initials: J
authors.lastname: Cruz
authors.forename: Laura
authors.initials: L
authors.lastname: Orphin
authors.forename: Pamela
authors.initials: P
authors.lastname: Pazoles
authors.forename: Mary Dawn T
authors.initials: MD
authors.lastname: Co
authors.forename: Francis A
authors.initials: FA
authors.lastname: Ennis
authors.forename: Masanori
authors.initials: M
authors.lastname: Terajima
mesh_terms: D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D003429:Cross Reactions; D003602:Cytotoxicity, Immunologic; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D016679:Genome, Viral / Q000276:immunology; D055106:Genome-Wide Association Study; D006388:Hemagglutinins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006650:Histocompatibility Testing; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D009439:Neuraminidase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
chemical_list: D018984:Epitopes, T-Lymphocyte; D006388:Hemagglutinins; C488936:M1 protein, Influenza A virus; D010446:Peptide Fragments; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D007371:Interferon-gamma; D009439:Neuraminidase