P1088Expression Signatures of Equid Lymphocyte Subsets Defined By Single Cell Sequencing
Expression Signatures of Equid Lymphocyte Subsets Defined By Single Cell Sequencing
Peripheral blood lymphocytes (PBL) from horse (E. caballus) and donkey (E. asinus) were analyzed in the 10X GenomicsTM CHROMIUMTM Single Cell 3’ Solution pipeline followed by library sequencing using a NextSeq500 instrument. From an input number of 3000 cells for each species single cell sequence was obtained for 1781 cells from horse (59%) and 1462 cells from donkey (49%). Counts of reads per cell were 68.3k for horse and 79.3k for donkey, with 90% of the reads mapping to the reference sequence for horse and 46% of reads mapping to Ensembl predicted gene models. Corresponding figures for donkey were 88% and 53%. The 10X GenomicsTM software provided visualization of data, quantification of expression in individual cells, and clustering that grouped cells with similar expression profiles. By examining differentially expressed genes we were able to identify clusters that correspond to the major B cell, CD4 T-cell, and CD8 T-cell subpopulations of horse and donkey PBL. The percentages of cells in the subpopulations correlated well with the same subsets defined using monoclonal antibodies. The single cell sequencing approach facilitates identification of heterogeneity of expression in complex cell populations that cannot be distinguished using conventional light microscopy or separated using immunological techniques, and thus should have many applications in domestic animal genomics. Quantitative analysis of expression in our samples is underway.