Gene expression analysis of total B, CD4+ T and epithelial cells in lacrimal glands from the NOD mouse model of Sjogren's syndrome. Mus musculus

Despite being one of the most common rheumatologic diseases, there is still no disease-modifying drug for primary Sjögren’s syndrome (pSS). Advancing our knowledge of the target tissue has been limited by the low dimensionality of histology techniques and the small size of human salivary gland biopsies. The goal of this study was to characterize the gene expression profile of total B cells, CD4+ T cells and epithelial cells in the target tissue of the most commonly-used model of pSS (the NOD model of pSS characterized by severe dacryoadenitis developing in males between 8 and 16 weeks of age). Overall design: All immune subsets were purified from lacrimal gland (LG) cell suspensions of 16-week old male NOD mice by FACS cell sorting (>99% purity) using the following surface markers: CD326, CD45, NKp46, CD11b, CD11c, B220, CD3, CD4 and DAPI as live/dead marker. Epithelial cells were isolated as CD45-CD326+ cells; CD4+ T cells were isolated as CD45+NKp46-CD11b-CD11c-B22-CD3+CD4+ cells; B cells were isolated as CD45+NKp46-CD11b-CD11c-B220+CD3- cells. RNA was purified and QC’d before processing for gene expression analysis. Splenic subsets were purified from 4-week old NOD mice as steady-state control immune cells to be compared with tissue-infiltrating subsets.