Gene expression analysis of skewed mouse T cells pre- vs post-injection, and ex vivo skin T cells from first disease vs flare, in a model of Cutaneous Lupus Erythematosus

We performed gene expression analysis on cultured Th0, Th1 and Th2 cells pre-injection, and on enriched T cells from lesional skin of cutaneous lupus erythematosus (CLE) mice post-injection, to compare differences in transcription that could predict skin tropism and flare DO.11 T cells were skewed in vitro towards Th0, Th1 or Th2. A portion of the cells were saved for RNA analysis and the remainder were injected into recipient mice. Only Th2 cells induced skin disease. To analyze gene expression post-injection, we enriched for T cells from lesional skin of mice in the first course of disease (approximately 4 weeks) or at flare (approximately 12 weeks) using Miltenyi CD3 bead isolation kits. All extracted RNA samples were analyzed using NanoString mouse Immunology panel (C2269).