Transcriptional modulation of TCR, notch and Wnt signaling pathways in SEB-anergized CD4+ T cells

Gene expression changes in CD4+V beta 8+ T cells anergized by in vivo exposure to staphylococcal enterotoxin B ( SEB) bacterial superantigen compared to CD4+V beta 8+ nonanergic T cells were assessed using DNA microarrays containing 5184 murine complementary DNAs. Anergy in splenic T cells of SEB-immunized BALB/c mice was verified by dramatically reduced proliferative capacity and an 8x overexpression of GRAIL mRNA in CD4+Vb8+ T cells taken from mice 7 days after injection. At an Associative t-test threshold of P < 0.0005, 96 genes were overexpressed or detected only in anergic T cells, while 256 genes were suppressed or not detected in anergic T cells. Six of eight differential expressions tested using real-time quantitative PCR were validated. Message for B-Raf was detected only in non-anergic cells, while expression of the TCR signaling modulator Slap (Src-like adapter protein) and the TCR zeta-chain specific phosphatase Ptpn3 was enhanced. Modulation of multiple genes suggests downregulation of Wnt/beta-catenin signaling and enhanced Notch signaling in the anergic cells. Consistent with previous reports in a non-superantigen in vivo anergy model, mRNA for CD18 and the transcription factor Satb1 (special AT-rich-binding protein 1) was increased in SEB-anergized T cells. This is the first report of global transcriptional changes in CD4+ T-cells made anergic by superantigen exposure.