Synthesis of chicken major histocompatibility complex class II oligomers

Chicken major histocompatibility complex (MHC) B21 and B19 haplotypes are associated with resistance and susceptibility to Marek's disease (MD), respectively. MHC-associated resistance may be related to differential immune response to Marek's disease virus in B21 and B19 chickens. In order to study T-cell response to MD virus in these haplotypes, the objective of this study was to synthesize chicken MHC class II oligomers. Full-length coding sequences of chicken MHC class II (MHC-II) B21 and B19 molecules were amplified and the molecules were expressed using a baculovirus expression system in insect cells. Recombinant MHC-II were exogenously loaded with naturally processed and presented peptides which stabilized the heterodimer in sodium dodecyl sulphate polyacrylamide gel electrophoresis and allowed the detection of these molecules in western blots with a conformation-specific anti-chicken MHC-II antibody. Biotinylated MHC molecules were conjugated to streptavidin to form oligomers, which were confirmed under the transmission electron microscope through immunogold labelling.