Details

Autor(en) / Beteiligte

• Meiring, Hugo D
• Kuipers, Betsy
• van Gaans-van den Brink, Jacqueline A. M
• Poelen, Martien C. M
• Timmermans, Hans
• Baart, Gino
• Brugghe, Humphrey
• van Schie, Joost
• Boog, Claire J. P
• de Jong, Ad P. J. M
• van Els, Cecile A. C. M

Titel

Mass Tag-Assisted Identification of Naturally Processed HLA Class II-Presented Meningococcal Peptides Recognized by CD4+ T Lymphocytes

Ist Teil von

• The Journal of immunology (1950), 2005-05, Vol.174 (9), p.5636-5643

Ort / Verlag

United States: Am Assoc Immnol

Erscheinungsjahr

2005

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 (14)N- and (15)N-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 ( approximately 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.