title: Physical detection of influenza A epitopes identifies a stealth subset on human lung epithelium evading natural CD8 immunity.
abstract: Vaccines eliciting immunity against influenza A viruses (IAVs) are currently antibody-based with hemagglutinin-directed antibody titer the only universally accepted immune correlate of protection. To investigate the disconnection between observed CD8 T-cell responses and immunity to IAV, we used a Poisson liquid chromatography data-independent acquisition MS method to physically detect PR8/34 (H1N1), X31 (H3N2), and Victoria/75 (H3N2) epitopes bound to HLA-A*02:01 on human epithelial cells following in vitro infection. Among 32 PR8 peptides (8-10mers) with predicted IC50 < 60 nM, 9 were present, whereas 23 were absent. At 18 h postinfection, epitope copies per cell varied from a low of 0.5 for M13-11 to a high of >500 for M1(58-66) with PA, HA, PB1, PB2, and NA epitopes also detected. However, aside from M1(58-66), natural CD8 memory responses against conserved presented epitopes were either absent or only weakly observed by blood Elispot. Moreover, the functional avidities of the immunodominant M1(58-66)/HLA-A*02:01-specific T cells were so poor as to be unable to effectively recognize infected human epithelium. Analysis of T-cell responses to primary PR8 infection in HLA-A*02:01 transgenic B6 mice underscores the poor avidity of T cells recognizing M1(58-66). By maintaining high levels of surface expression of this epitope on epithelial and dendritic cells, the virus exploits the combination of immunodominance and functional inadequacy to evade HLA-A*02:01-restricted T-cell immunity. A rational approach to CD8 vaccines must characterize processing and presentation of pathogen-derived epitopes as well as resultant immune responses. Correspondingly, vaccines may be directed against "stealth" epitopes, overriding viral chicanery.
authors: Department of Medical Oncology, Laboratory of Immunobiology, Cancer Vaccine Center, Dana-Farber Cancer Institute, and Department of Medicine, Harvard Medical School, Boston, MA 02115;
authors: Derin B
authors: DB
authors: Keskin
authors: Department of Medical Oncology, Laboratory of Immunobiology, Cancer Vaccine Center, Dana-Farber Cancer Institute, and Department of Medicine, Harvard Medical School, Boston, MA 02115;
authors: Bruce B
authors: BB
authors: Reinhold
authors: Cancer Vaccine Center, Dana-Farber Cancer Institute, and Department of Medicine, Harvard Medical School, Boston, MA 02115; Computer Science Department, Metropolitan College, Boston University, Boston, MA 02115; and.
authors: Guang Lan
authors: GL
authors: Zhang
authors: Barnett Institute, Northeastern University, Boston, MA 02115.
authors: Alexander R
authors: AR
authors: Ivanov
authors: Barnett Institute, Northeastern University, Boston, MA 02115.
authors: Barry L
authors: BL
authors: Karger
authors: Department of Medical Oncology, Laboratory of Immunobiology, Cancer Vaccine Center, Dana-Farber Cancer Institute, and Department of Medicine, Harvard Medical School, Boston, MA 02115; ellis_reinherz@dfci.harvard.edu.
authors: Ellis L
authors: EL
authors: Reinherz
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000276:immunology / Q000821:virology; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006801:Humans; D009980:Influenza A virus / Q000276:immunology*; D008168:Lung / Q000276:immunology / Q000821:virology*
chemical_list: D000939:Epitopes
keywords: T-cell avidity; T-cell epitopes; antigen presentation; influenza A viruses; mass spectrometry
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