Ovsyannikova Inna G, Johnson Kenneth L, Naylor Stephen, Poland Gregory A
Mayo Vaccine Research Group, 611C Guggenheim Building, Mayo Clinic, 200 First Street SW, Rochester, Minnesota 55905, USA.
J Immunol Methods. 2005 Feb;297(1-2):153-67. doi: 10.1016/j.jim.2004.12.020.
We developed a B-lymphocyte cell line derived from a measles seropositive individual who was homozygous for the HLA-DRB10301 allele. Peptides associated with the HLA-DRB10301 protein were purified from this lymphoblastoid cell line after infection with the Attenuvax measles vaccine virus (Merck Research Laboratories, West Point, PA) and with "sham" infection. More than 40 peptide sequences were obtained by nano-scale reversed phase-high performance liquid chromatography coupled to tandem mass spectrometry (nano-LC/MS/MS). These peptides originated from 21 different source proteins--the majority from membrane-bound proteins. Most of the peptides (>73%) bound to HLA-DRB10301 appeared to be in lower abundance on measles-infected cells compared to the "sham-infected" cells. However, 26% of the identified peptides seem to have increased expression after measles infection. Measles vaccine virus infection did not change the level of HLA-DR expression. We demonstrate the power of nano-LC/MS/MS in the rapid determination of changes in the spectrum and expression of HLA-DRB10301-bound peptides after infection with measles virus. This provides further knowledge of the changes in peptide expression after viral infection and provides a powerful tool for identifying HLA-presented host and viral epitopes in the context of class II HLA molecules.
