RESUMEN
We have designed a cytometry-based competition assay to evaluate peptide binding to empty recombinant HLA class II molecules. The efficiency of this assay was evaluated using recombinant HLA-DP0401 molecules (HLA-DP) produced in insect cells and 13 peptides from human telomerase reverse transcriptase (hTERT). We demonstrate that our method allowed accurate measurements of peptide Ki values and can thus discriminate strong, moderate and poor HLA-DP binders. In parallel, we showed that among hTERT peptides, the most immunodominant in healthy individuals were those with moderate affinity for HLA-DP while no T cell response could be evidenced against peptides with very strong or very low affinities for HLA-DP. This strongly suggests that the precise determination of peptide affinity with our method can improve HLA class II epitope prediction.
Asunto(s)
Antígenos de Histocompatibilidad Clase II/metabolismo , Inmunoensayo/métodos , Péptidos/inmunología , Péptidos/metabolismo , Secuencia de Aminoácidos , Animales , Unión Competitiva , Biotinilación , Línea Celular , Drosophila , Citometría de Flujo/métodos , Antígenos HLA-DP/genética , Antígenos HLA-DP/metabolismo , Cadenas alfa de HLA-DP , Cadenas beta de HLA-DP , Antígenos de Histocompatibilidad Clase II/genética , Humanos , Datos de Secuencia Molecular , Péptidos/administración & dosificación , Péptidos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Linfocitos T/inmunología , Telomerasa/genética , Telomerasa/inmunología , Telomerasa/metabolismoRESUMEN
Alloy Pt-M (M = Co, Ni) nanocatalysts, supported on carbon Vulcan XC-72, were synthesized using the carbonyl chemical route. A high dispersion on such substrate was revealed by transmission electron microscopy (TEM). Alloy formation on the nanometre scale length was shown by high-resolution transmission microscopy (HRTEM) and energy dispersive X-ray spectroscopy (EDX) on a nanoparticle. The metal M in Pt-M nanoalloys segregates preferentially on the nanoparticles' surface, as determined by the hydrogen adsorption electrochemical reaction. An increased tolerance towards methanol of such nanoalloy materials was observed for the oxygen reduction reaction (ORR) in acid media. To better understand the structure and segregation phenomena of these nanoalloys, molecular dynamics (MD) with a self-optimized reactive force field was applied.