Intranuclear Delivery of HIF-1α-TMD Alleviates EAE via Functional Conversion of TH17 Cells

T helper 17 (TH17) cells are involved in several autoimmune diseases such as multiple sclerosis (MS) and rheumatoid arthritis (RA). In addition to retinoic acid receptor-related orphan nuclear receptor gamma t (ROR-gamma t), hypoxia-inducible factor-1 alpha (HIF-1 alpha) is essential for the differentiation and inflammatory function of TH17 cells. To investigate the roles of HIF-1 alpha in the functional regulation of TH17 cells under the normal physiological condition without genetic modification, the nucleus-transducible form of transcription modulation domain (TMD) of HIF-1 alpha (ntHIF-1 alpha-TMD) was generated by conjugating HIF-1 alpha-TMD to Hph-1 protein transduction domain (PTD). ntHIF-1 alpha-TMD was effectively delivered into the nucleus of T cells without cellular cytotoxicity. ntHIF- 1 alpha-TMD significantly blocked the differentiation of naive T cells into TH17 cells in a dose-dependent manner via IL-17A and ROR-gamma t expression inhibition. However, T- cell activation events such as induction of CD69, CD25, and IL-2 and the differentiation potential of naive T cells into TH1, TH2, or Treg cells were not affected by ntHIF-1 alpha-TMD. Interestingly, TH17 cells differentiated from naive T cells in the presence of ntHIF-1 alpha-TMD showed a substantial level of suppressive activity toward the activated T cells, and the increase of Foxp3 and IL-10 expression was detected in these TH17 cells. When mRNA expression pattern was compared between TH17 cells and ntHIF-1 alpha-TMD-treated TH17 cells, the expression of the genes involved in the differentiation and functions of TH17 cells was downregulated, and that of the genes necessary for immune-suppressive functions of Treg cells was upregulated. When the mice with experimental autoimmune encephalomyelitis (EAE) were treated with ntHIF-1 alpha-TMD with anti-IL-17A mAb as a positive control, the therapeutic efficacy of ntHIF-1 alpha-TMD in vivo was comparable with that of anti-IL-17A mAb, and ntHIF-1 alpha-TMD-mediated therapeutic effect was contributed by the functional conversion of TH17 cells into immune-suppressive T cells. The results in this study demonstrate that ntHIF-1 alpha-TMD can be a new therapeutic reagent for the treatment of various autoimmune diseases in which TH17 cells are dominant and pathogenic T cells.