Effects of HLA DRB1*15:01 interactions with ?-syn32-46 on gene expression in intestinal immune cells in mice
HLA DRB1*15:01 is overrepresentated in Parkinson's disease patients and binds with high affinity to the ?-synuclein peptide, 32-46. Immunization of humanized mice expressing HLA DRB1*15:01 with ?-syn32-46 induces enteric phenotypes similar to those of prodromal Parkinson's disease. We collected the small intestines from HLA mice immunized with either Complete Freund's Adjuvant (CFA) with ?-syn32-46 or CFA alone, sorted the CD45+ immune cells, then performed single-cell RNA sequencing to evaluate gene expression at single cell resolution. Overall design: 10-12 week old HLA DRB1*15:01 expressing MHC-II (-/-) mice were immunized with ?-syn32-46 (100ug) in CFA (200ug) or CFA alone. At 14 days post immunization (DPI), mice were boosted again with the same emulsion followed by an injection of pertussis toxin (400ng), then an additional pertussis toxin injection at 16 DPI. At 21 DPI, the small intestine was collected and dissociated to single cells. The cells were sorted, and CD45+ cells were collected for single-cell RNA sequencing.