T-helper cell type 17 (Th17) mediated inflammation is associated with various diseases including autoimmune encephalitis, inflammatory bowel disease and lung diseases such as chronic obstructive pulmonary disease and asthma. Differentiation into distinct T helper subtypes needs to be tightly regulated to ensure an immunological balance. As microRNAs (miRNAs) are critical regulators of signalling pathways, we aimed to identify specific miRNAs implicated in controlling Th17 differentiation. We were able to create a regulatory network model of murine T helper cell differentiation by combining Affymetrix mRNA and miRNA arrays and in silico analysis. In this model, the miR-212~132 and miR-182~183 clusters were significantly up-regulated upon Th17 differentiation, whereas the entire miR-106~363 cluster was down-regulated and predicted to target well-known Th17 cell differentiation pathways. In vitro transfection of miR-18b, miR-106a and miR-363-3p into primary murine Cd4(+) lymphocytes decreased expression of retinoid-related orphan receptor c (Rorc), Rora, Il17a and Il17f, and abolished secretion of Th17-mediated interleukin-17a (Il17a). Moreover, we demonstrated target site-specific regulation of the Th17 transcription factors Rora and nuclear factor of activated T cells (Nfat) 5 by miR-18b, miR-106a and miR-363-3p through luciferase reporter assays. Here, we provide evidence that miRNAs are involved in controlling the differentiation and function of T helper cells, offering useful tools to study and modify Th17-mediated inflammation.
- Kastle, M.
- Bartel, S.
- Geillinger-Kastle, K.
- Irmler, M.
- Beckers, J.
- Ryffel, B.
- Eickelberg, O.
- Krauss-Etschmann, S.
Keywords
- Animals
- *Cell Differentiation
- Cells, Cultured
- Female
- Gene Expression Profiling/methods
- Gene Expression Regulation
- Gene Regulatory Networks
- Interleukin-17/genetics/metabolism
- Mice, Inbred BALB C
- MicroRNAs/genetics/*metabolism
- Nuclear Receptor Subfamily 1, Group F, Member 1/genetics/metabolism
- Nuclear Receptor Subfamily 1, Group F, Member 3/genetics/metabolism
- Oligonucleotide Array Sequence Analysis
- Phenotype
- Signal Transduction
- Th17 Cells/immunology/*metabolism
- Th2 Cells/immunology/metabolism
- Transcription Factors/genetics/metabolism
- Transfection
- *T helper cell differentiation
- *T helper type 17
- *miR-106a
- *miR-18b
- *miR-363a-3p
- *microRNA
- *nuclear factor of activated T cells 5
- *retinoid-related orphan receptor alpha
- *retinoid-related orphan receptor gammat