Crinophagic granules in pancreatic ß cells contribute to mouse autoimmune diabetes by diversifying pathogenic epitope repertoire
We examined the transcriotional changes in antigen specific CD4 T cells to antigens derived from thymus or to the antigens derived from islets in the autoimmune diabetes mouse model. Overall design: Single cells were prepared from spleen and lymph nodes of 8-12 weeks old NOD mice or HEL specfic TCR transgenic mice. Tetramer enriched cells were sorted from the NOD mice and naïve CD4 T cells were sorted from HEL TCR trangenic mice. Before surface staining, samples were incubated with 100 µl of TotalSeq™ anti-mouse Hashtag reagent (1:100 [vol/vol], BioLegend) to tag the samples: enriched cells from the islet epitope set were tagged with the TotalSeq-C0301 anti-mouse Hashtag 1 Antibody (Barcode sequence: ACCCACCAGTAAGAC); enriched cells from the thymic epitope set were tagged with the TotalSeq-C0302 anti-mouse Hashtag 2 Antibody (Barcode sequence: GGTCGAGAGCATTCA); 10E11 naïve CD4+ T cells were tagged with the TotalSeq-C0303 anti-mouse Hashtag 3 Antibody (Barcode sequence: CTTGCCGCATGTCAT).