dc.contributor |
Universitat de Vic - Universitat Central de Catalunya. Càtedra de la Sida i Malalties Relacionades |
dc.contributor.author |
Yaciuk, Jane C. |
dc.contributor.author |
Skaley, Matthew |
dc.contributor.author |
Bardet, Wilfried |
dc.contributor.author |
Schafer, Danijela Mojsilovic |
dc.contributor.author |
Cate, Steven |
dc.contributor.author |
Stewart, Christopher J. |
dc.contributor.author |
McMurtrey, Curtis |
dc.contributor.author |
Jackson, Rico Buchli |
dc.contributor.author |
Olvera, Alex |
dc.contributor.author |
Cedeño, Samandhy |
dc.contributor.author |
Plana, Montserrat |
dc.contributor.author |
Mothe, B. |
dc.contributor.author |
Brander, Christian |
dc.contributor.author |
West, John T. |
dc.contributor.author |
Hildebrand, William |
dc.date |
2014 |
dc.identifier |
Yaciuk, J. C., Skaley, M., Bardet, W., Schafer, F., Mojsilovic, D., Cate, S., et al. (2014). Direct interrogation of viral peptides presented by the class I HLA of HIV-infected T cells. Journal of Virology, 88(22), 12992-13004. |
dc.identifier |
0022-538X |
dc.identifier |
http://hdl.handle.net/10854/3578 |
dc.identifier |
https://doi.org/ 10.1128/JVI.01914-14 |
dc.identifier.uri |
http://hdl.handle.net/10854/3578 |
dc.description |
Identification of CD8+ cytotoxic T lymphocyte (CTL) epitopes has traditionally relied upon testing of overlapping peptide libraries for their reactivity with T cells in vitro. Here, we pursued deep ligand sequencing (DLS) as an alternative method of directly identifying those ligands that are epitopes presented to CTLs by the class I human leukocyte antigens (HLA) of infected cells. Soluble class I HLA-A*11:01 (sHLA) was gathered from HIV-1 NL4-3-infected human CD4+ SUP-T1 cells. HLA-A*11:01 harvested from infected cells was immunoaffinity purified and acid boiled to release heavy and light chains from peptide ligands that were then recovered by size-exclusion filtration. The ligands were first fractionated by high-pH high-pressure liquid chromatography and then subjected to separation by nano-liquid chromatography (nano-LC)–mass spectrometry (MS) at low pH. Approximately 10 million ions were selected for sequencing by tandem mass spectrometry (MS/MS). HLA-A*11:01 ligand sequences were determined with PEAKS software and confirmed by comparison to spectra generated from synthetic peptides. DLS identified 42 viral ligands presented by HLA-A*11:01, and 37 of these were previously undetected. These data demonstrate that (i) HIV-1 Gag and Nef are extensively sampled, (ii) ligand length variants are prevalent, particularly within Gag and Nef hot spots where ligand sequences overlap, (iii) noncanonical ligands are T cell reactive, and (iv) HIV-1 ligands are derived from de novo synthesis rather than endocytic sampling. Next-generation immunotherapies must factor these nascent HIV-1 ligand length variants and the finding that CTL-reactive epitopes may be absent during infection of CD4+ T cells into strategies designed to enhance T cell immunity. |
dc.format |
application/pdf |
dc.language |
eng |
dc.publisher |
American Socity for Microbiology |
dc.relation |
http://jvi.asm.org/content/88/22/12992.abstract |
dc.rights |
(c) American Society for Microbiology |
dc.rights |
Tots els drets reservats |
dc.rights |
info:eu-repo/semantics/openAccess |
dc.subject |
Sida -- Tractament |
dc.title |
Direct Interrogation of Viral Peptides Presented by the Class I HLA of HIV-Infected T Cells |
dc.type |
info:eu-repo/semantics/article |
dc.type |
info:eu-repo/publishedVersion |