Mass tag-assisted identification of naturally processed HLA class II-presented meningococcal peptides recognized by CD4+ T lymphocytes
| Autor: | Ad P. J. M. de Jong, Claire J. P. Boog, Betsy Kuipers, Jacqueline A. M. van Gaans-van den Brink, Humphrey F. Brugghe, Gino J E Baart, Cécile A. C. M. van Els, Martien C. M. Poelen, Joost van Schie, Hans A. M. Timmermans, Hugo D. Meiring |
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| Rok vydání: | 2005 |
| Předmět: |
CD4-Positive T-Lymphocytes
Nitrogen T cell Immunology Molecular Sequence Data Epitopes T-Lymphocyte Porins Meningococcal Vaccines Neisseria meningitidis Epitope Microbiology MHC class I medicine Immunology and Allergy Humans Amino Acid Sequence Peptide sequence Cells Cultured MHC class II Antigen Presentation biology Nitrogen Isotopes Immunodominant Epitopes Homozygote HLA-DR1 Antigen Dendritic Cells Molecular biology Peptide Fragments medicine.anatomical_structure Amino Acid Substitution Porin biology.protein Bacterial outer membrane Memory T cell Bacterial Outer Membrane Proteins |
| Zdroj: | Journal of immunology (Baltimore, Md. : 1950). 174(9) |
| ISSN: | 0022-1767 |
| Popis: | The meningococcal class I outer membrane protein porin A plays an important role in the development of T cell-dependent protective immunity against meningococcal serogroup B infection and is therefore a major component of candidate meningococcal vaccines. T cell epitopes from porin A are poorly characterized because of weak in vitro memory T cell responses against purified Ag and strain variation. We applied a novel strategy to identify relevant naturally processed and MHC class II-presented porin A epitopes, based on stable isotope labeling of Ag. Human immature HLA-DR1-positive dendritic cells were used for optimal uptake and MHC class II processing of 14N- and 15N-labeled isoforms of the neisserial porin A serosubtype P1.5–2,10 in bacterial outer membrane vesicles. HLA-DR1 bound peptides, obtained after 48 h of Ag processing, contained typical spectral doublets in mass spectrometry that could easily be assigned to four porin A regions, expressed at diverging densities (∼30–4000 copies/per cell). Epitopes from two of these regions are recognized by HLA-DR1-restricted CD4+ T cell lines and are conserved among different serosubtypes of meningococcal porin A. This mass tag-assisted approach provides a useful methodology for rapid identification of MHC class II presented bacterial CD4+ T cell epitopes relevant for vaccine development. |
| Databáze: | OpenAIRE |
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