TY - JOUR ID - 250229 TI - Positive and Negative Regulation of Th17 Cell Differentiation: Evaluating The Impact of RORC2 JO - Cell Journal (Yakhteh) JA - CELLJ LA - en SN - 2228-5806 AU - Ganjalikhani Hakemi, Mazdak AU - Ghaedi, Kamran AU - Homayouni, Vida AU - Andalib, Alireza AU - Hosseini, Mohsen AU - Rezaei, Abbas AD - Cellular and Molecular Immunology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran;Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran AD - Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran AD - Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran AD - 4Department of Epidemiology, Faculty of Health, Isfahan University of Medical Sciences, Isfahan, Iran AD - Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran;5Applied Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran Y1 - 2014 PY - 2014 VL - 16 IS - 3 SP - 343 EP - 352 KW - siRNA KW - Th17 DO - N2 - ObjectiveTh17 cells are known to be involved in some types of inflammations and autoimmune disorders. RORC2 is the key transcription factor coordinating Th17 cell differentiation. Thus, blocking RORC2 may be useful in suppressing Th17-dependent inflammatory processes. The aim was to silence RORC2 by specific siRNAs in naïve T cells differentiating to Th17. Time-dependent expression of RORC2 as well as IL-17 and IL-23R were considered before and after RORC2 silencing. Materials and Methods In this experimental study, naïve CD4+T cells were isolated from human cord blood samples. Cytokines TGFß plus IL-6 and IL-23 were used to polarize the naïve T cells to Th17 cells in X-VIVO 15 serum free medium. A mixture of three siRNAs specific for RORC2 was applied for blocking its expression. RORC2, IL-17 and IL-23R mRNA and protein levels were measured using qRT-PCR, ELISA and flow cytometry techniques. Pearson correlation and one-way ANOVA were used for statistical analyses. Results Significant correlations were obtained in time-dependent analysis of IL-17 and IL-23R expression in relation with RORC2 (R=0.87 and 0.89 respectively, p < 0.05). Silencing of RORC2 was accompanied with almost complete suppression of IL-17 (99.3%; p < 0.05) and significant decrease in IL-23R gene expression (77.2%, p < 0.05). Conclusion Our results showed that RORC2 is the main and the primary trigger for upregulation of IL-17 and IL-23R genes in human Th17 cell differentiation. Moreover, we show that day 3 could be considered as the key day in the Th17 differentiation process. UR - https://www.celljournal.org/article_250229.html L1 - https://www.celljournal.org/article_250229_8b0f385908523ea37a3c2540a88a9d27.pdf ER -