MicroRNA-146a expresses in interleukin-17 producing T cells in rheumatoid arthritis patients
© Niimoto et al; licensee BioMed Central Ltd. 2010
Received: 24 May 2010
Accepted: 15 September 2010
Published: 15 September 2010
Interleukin (IL)-17 is an important factor in rheumatoid arthritis (RA) pathogenesis. MicroRNA (miRNA)s are a family of non coding RNAs and associated with human diseases including RA. The purpose of this study is to identify the miRNAs in the differentiation of IL-17 producing cells, and analyze their expression pattern in the peripheral blood mononuclear cells (PBMC) and synovium from RA patients.
IL-17 producing cells were expanded from CD4+T cell. MiRNA microarray was performed to identify the miRNAs in the differentiation of IL-17 producing cells. Quantitative polymerase chain reaction was performed to examine the expression patterns of the identified miRNAs in the PBMC and synovium from RA and osteoarthritis (OA) patients. Double staining combining in situ hybridization and immunohistochemistry of IL-17 was performed to analyze the expression pattern of identified miRNA in the synovium.
Six miRNAs, let-7a, miR-26, miR-146a/b, miR-150, and miR-155 were significantly up regulated in the IL-17 producing T cells. The expression of miR-146a and IL-17 was higher than in PBMC in the patients with low score of Larsen grade and short disease duration. MiR-146a intensely expressed in RA synovium in comparison to OA. MiR-146a expressed intensely in the synovium with hyperplasia and high expression of IL-17 from the patients with high disease activity. Double staining revealed that miR-146a expressed in IL-17 expressing cells.
These results indicated that miR-146a was associated with IL-17 expression in the PBMC and synovium in RA patients. There is the possibility that miR-146a participates in the IL-17 expression.
Rheumatoid arthritis (RA) is characterized by chronic synovial inflammation and subsequent joint destruction . The infiltration of macrophages, T cells and B cells in RA synovium plays a crucial role in RA pathogenesis including proliferation of the lining cells, and production of inflammatory cytokines such as tumor necrosis factor- (TNF-)α, and interleukin-1- (IL-1-)β. However, the pathogenesis of RA has not been completely elucidated.
The discovery of a new linage of CD4+ effector T helper type17 cells (Th17cells) that selectively produce IL-17 has provided exciting new insights into immune regulation, host defense, and pathogenesis of autoimmune and other chronic inflammatory disorders including of RA [2–4]. IL-17 is a proinflammatory cytokine, which induces other cytokines, such as TNFα, IL-1β, IL-6, IL-23 and G-CSF [5–8]. In addition, IL-17 plays a role in osteoclastogenesis via activation of RANKL(receptor activator of NF-κB ligand), causing bone destruction in inflammatory joints [9–11]. Several studies demonstrated that IL-17 is higher in synovial fluid, synovium and peripheral blood mononuclear cells in RA patients than that in healthy subjects [12–14]. IL-17 is recognized to be the one of the important factors in RA pathogenesis.
MicroRNA (miRNA)s are a family of ~22-nucleotide non coding RNAs identified in organisms ranging from nematodes to humans [15–17]. Many miRNAs are evolutionarily conserved across phyla, thereby regulating gene expression by posttranscriptional gene repression. The miRNAs regulate gene expression by binding the 3'-untranslated region of their target mRNAs leading to translational repression or mRNA degradation [18–21]. Several microRNAs exhibit a tissue-specific or developmental stage-specific expression pattern and have been reported to be associated with human diseases such as cancer, leukemia, and viral infection [22, 23]. These findings suggest their potential as a novel therapeutic target. miRNA might play a role in RA pathogenesis in autoimmune and other chronic inflammatory diseases including of RA. Several studies reported that miRNA might play a role in RA pathogenesis. Stancyzk et al. reported that miR-146 and miR-155 are highly expressed in RA synovial fibroblast in comparison to osteoarthritis fibroblast . Nakasa et al. demonstrated that miR-146 is highly expressed in RA synovial tissue in comparison to OA and normal synovial tissue. They also revealed that miR-146 is expressed primarily in CD68+ macrophages, but also in some CD3+ T cell subsets and CD79a+ B cells in RA synovial tissue . Paulay et al. reported that PBMCs from RA patients exhibit statistically significant increase the expression levels of miR-146a, miR-155, miR-132, and miR-16 in comparison to healthy and disease control individuals. They also demonstrated that high levels of miR-146a and miR-16 expression correlate with active disease, whereas low expression levels correlate with inactive disease . Changsheng Du et al. reported that miR-326 regulates Th-17 differentiation and associated with the pathogenesis of multiple sclerosis. These findings suggest that miRNA also might play a role in the expression of IL-17, and an analysis of the expression pattern of miRNAs in IL-17 producing T cells might lead to the development of new treatments for RA.
The purpose of this study is to identify miRNAs in the differentiation of IL -17 producing T cells, and to analyze their expression pattern in the RA patients. MiRNAs were identified during the differentiation of IL-17 producing T cells by the expansion from healthy CD4+ T cells using microarray analysis, and analyzed the expression pattern of the identified miRNAs in the peripheral blood mononuclear cells (PBMC) and synovium from RA and OA patients.
Demographic and clinical features of this study subjects.
disease duration Larsen grade K/L score
MTX, predonisone, NSAIDs
MTX, predonisone, NSAIDs
MTX, bucillamine, predonisone, NSAIDs
Cell isolation and expansion of IL-17 producing T cells
This clinical study was approved by the University of Hiroshima Institutional Review Board, and written permission was obtained from all healthy volunteers who participated in this study. Human peripheral blood was collected from 5 healthy volunteers (31.8 ± 1.1 years of age, mean ± SD) into DPBS-E (5 mM: 0.5 M EDTA) drop by drop and mixed well. This mixture was loaded slowly onto Histopaque® (Sigma Chemical Co. CA) in another tube, and centrifuged at 1000 × g at room temperature for 10 min. PBMC accumulated as the middle white monolayer. After the supernatant was discarded, only the white monolayer cells were aspirated and put into DPBS-E, then centrifuged at 400 × g at 4°C for 10 min. The supernatant was discarded, DPBS-E and ammonium chloride (The cell Experts™) were added at the rate of 1:3, and the mixture was allowed to stand at room temperature for 10 min. After centrifuging at 400 × g at 4°C for 10 min, supernatant was discarded, and DPBS-E was added and mixed well. This final process was repeated several times. The remaining white cells were peripheral blood mononuclear cells (PBMC). CD4+ T cells were isolated from PBMC using auto MACS (CD4+T cell Isolation kit, Miltenyi Biotec,). IL-17 producing cells were expanded from CD4+ T cells as previously described . PBMC or purified CD4+ T cells (1 × 106 per ml) were re-suspend in fresh culture medium containing requisite antibiotics and serum plus IL-2 (100 u/ml), IL -1 β, IL-2, IL -6 and IL -23 (R&D Systems; 10ng/ml). The cells were cultured in 24 well plates (2 ml per well) at 37°C for 4 days in 5% CO2. Afterwards, IL-17 expression was confirmed with RT-PCR and ELISA, and the presence of IL-17 producing T cells was also confirmed (data is not shown).
Three synovial tissue specimens per one patient were obtained from random sites during surgery. Each was visually inspected to minimize contamination with non-inflammatory tissue. Tissues were stored at -70°C until analysis. Total RNA for the PCR analysis was isolated from tissues homogenized with Trizol (Invitrogen) on ice for. The tissue specimens were fixed in 4% paraformaldehyde and paraffin-embedded for histopathological analysis.
miRNA microarrays (NCode microarray, Invitrogen) were performed for the identification of miRNA in differentiating IL-17 producing cells. The dual-color dye swap method was used to analyze differences of miRNA expression between expanded IL-17 producing cells and non-expanded cells. Five-hundred nanograms of the enriched miRNA was labeled with the NCode Rapid miRNA labeling kit (Invitrogen) and hybridized to NCode multispecies miRNA arrays as described earlier based on a loop design, balanced with respect to array, dye, and sample. The balanced design minimized sources of unwanted variation. The arrays were scanned, aligned, and median spot intensities were obtained using a GenePix 4000B scanner (Molecular Devices, Inc., Sunnyvale, CA, USA).
Synthesis of complementary DNA
One microgram of total RNA was reverse-transcribed using the QuantiTect® Reverse Transcription Kit (Qiagen, Chatsworth, CA) according to the manufacturer's protocol. The genomic DNA elimination reaction was carried out using 2 μl of gDNA wipeout buffer, 1 μg (1 μl) template RNA and 11 μl RNase-free water at 42°C for 2 min. Reverse transcription was performed in 1 μl quantiscript reverse transcriptase, 4 μl quantiscript RT buffer, 1 μl RT primer mix and 14 μl template RNA (the entire genomic DNA elimination reaction) at 42°C for 15 min and 95°C for 3 min and then the cDNA product was maintained at 4°C.
Quantitative (real time) PCR
Quantitative RT-PCR assays were performed using a TaqMan miRNA assay kit (Applied Biosystems, CA, USA) for the expression of miRNAs and SYBR Green (Invitrogen) for the expression of IL-17, Foxp3, retinoid-related orphan receptor γt (RORγt), IL-1 receptor-associated kinase 1(IRAK1) and suppressor of cytokine signaling 1(SOCS1). Reverse transcriptase reactions of mature miRNAs contained a sample of total RNA, 50 nM stem-loop RT primer, 10 × RT buffer, 100 mM each dNTPs, 50 U/μl MultiScribe reverse transcriptase, and 20 U/μl RNase inhibitor. 15 μl reactions were incubated in a thermo cycler (BioRad) for 30 min at 16°C, 30 min at 42°C, 5 min at 85°C, and held at 4°C. Real time PCR was performed using a Mini Opticon Real-time PCR System (BioRad, Hercules, CA) in a 10 μl PCR mixture containing 1.33 μl RT product, 2 × TaqMan Universal PCR Master Mix, 0.2 μM TaqMan probe, 15 μM forward primer, and 0.7 μM reverse primer. Each SYBR Green reaction was performed with 1.0 μl template cDNA, 10 μl SYBR Green mix, 1.5 μM primer, and water to adjust the final volume to 20 μl. Primer sequences were: IL-17, 5'- AAG ACC TCA TTG GTG TCA CTG CT AC-3'(forward), 5'- ATC TCT CAG GGT CCT CAT TGC G-3' (reverse); Foxp3, 5'- GAG AAG CTG AGT GCC ATG CA -3'(forward), 5'- AGA GCC CTT GTC GGA TGA T -3'(reverse); RORγt, 5'- TGA GAA GGA CAG GGA GCC AA-3'(forward), 5'- CCA CAG ATT TTG CAA GGG ATC A -3' (reverse); SOCS1, 5'- GAA CTG CTT TTT CGC CCT TA -3'(forward), 5'- CTC GAA GAG GCA GTC GAA G -3'(reverse); IRAK1, 5'- GCT CTT TGC CCA TCT CTT TG -3'(forward), 5'- GCT ACC ACG CCA GGC TAA TA -3'(reverse); GAPDH, 5'- AAG AAT TGC AAG TCT ACA TAT CAC CCA AG. -3'(forward), 5'- GGT CAT GGT CAC AGA GCC ACC-3'(reverse). All reactions were incubated in a 48 well plate at 95°C for 10 min, followed by 40 cycles of 95°C for 15 seconds, and 60°C for 1 min and performed in triplicate. The U18 or GAPDH gene was used as a control to normalize any differences in the total RNA levels in each sample. A threshold cycle (CT) was observed in the exponential phase of amplification, and quantification of relative expression levels was performed using standard curves for target genes and the endogenous control. Geometric means were used to calculate the ΔΔCT (delta-delta CT) values and expressed as 2-ΔΔCT. The value of each control sample was set at 1 and was used to calculate the fold-change of target genes.
Ten μg of the protein were separated on NuPAGE® Novex® Bis-Tris Mini Gels (Invitrogen, Carlsband, CA) and transferred onto a nitrocellulose membrane (Invitrogen, Carlsband, CA). Mouse monoclonal antibody against a partial recombinant IRAK1 (Abnova, Taiwan) and rabbit polyclonal anti-actin antibody (Santa Cruz Biotechnology, Santa Cruz, CA) were used as primary antibodies. Anti-mouse goat IgG (MP Biomedicals, LLC, Santa Ana, CA) for IRAK1 and anti-rabbit goat IgG (MP Biomedicals, LLC, Santa Ana, CA) for actin were used for secondary antibodies. Band detection was performed using the enhanced chemiluminescence reagent, ECL Western Blotting Detection Reagents (GE Healthcare UK Ltd, Little Chalfont, Buckinghamshire).
Paraffin sections were deparaffinized through xylene for three changes of five minutes each, followed by graded alcohol immersions to water and phosphate buffered saline solutions, the sections were treated with the retrieval solutions (DAKO, Dakocytomation Inc.,Copinteria,CA,USA) for 20 minutes at 95°C. Next, the sections were depleted of endogenous peroxidase by incubation with 0.3% H2O2 in absolute methanol for 15 minutes. After blocking nonspecific biding with blocking reagent for 30 minutes, the sections incubated with primary antibody at appropriate dilutions for overnight at 4°C. For primary antibodies, monoclonal rabbit anti-human antibody against IL-17 (Santa Cruz Biotechnology,Inc,CA,USA). The sections were washed and incubated with biotinylated goat anti-mouse (Sigma, Saint Louis, Missouri, USA) for 1 hour at room temperature, then, washed and incubated with avidin-biotinylated horseradish peroxidase complex (ABC) and diaminobenzidine tetrahydrochloride (DAB; Dakocytomation Inc.,Copinteria,CA), and counterstained with Mayer's hematoxylin. The negative control was prepared in the same manner except that the primary antibody was omitted.
Double staining combining in situ hybridization and immunohistochemistry
After deparaffinization, each section was fixed in 4% paraformaldehyde for 10 minutes at room temperature, washed 3 times in phosphate buffered saline (PBS) for 3 minutes, and subsequently treated with 600 g of proteinase K for 10 minutes at room temperature. After treatment in 0.2% glycine-PBS for 10 minutes, the sections were refixed in 4% paraformaldehyde for 10 minutes, washed 3 times in PBS for 3 minutes each, and acetylated with 0.25% acetic anhydride in 0.1M triethanolamine hydrochloride for 10 minutes. After washing in PBS for 30 minutes, sections were prehybridized for 1 hour at 65°C with prehybridization buffer (50% formamide and 5 saline-sodium citrate [SSC]). Hybridization with DIG-labeled riboprobes of miR-146a (B-Bridge International, Mountain View, CA) was performed overnight at 65°C in hybridization buffer (50% formamide, 5X SSC, 5X Denhardt's solution, and 250 g/ml of Baker's yeast transfer RNA). After hybridization, sections were washed in 5X SSC for 30 minutes at 65°C, 0.2 SSC for 2 hours at 65°C, and 0.2X SSC for 5 minutes at room temperature. Blocking was performed overnight at 4°C with 4% horse serum and alkaline phosphatase-conjugated Fab anti-DIG antibody (Roche) in 1% sheep serum. Staining was performed using BCIP and nitroblue tetrazolium (NBT; Roche). Thereafter, the sections were washed in PBS and then were treated for 20 minutes at 90°C with retrieval solutions (Dakocytomation Inc.,Copinteria,CA). After blocking for 30 minutes, the sections were incubated with primary antibody of IL-17 at appropriate dilutions for 1 hour at room temperature. After washing, the sections were incubated with Alexa Fluor 594 conjugate(Invitrogen, Carlsbad, CA) for 30 minutes at room temperature, washed, and then incubated with 4, 6-diamidino-2-phenylindole (Dojindo Laboratories, Kumamoto, Japan). The negative control was prepared in the same manner, but without the primary antibody.
The Mann-Whitney U test was used to compare the gene expression between two groups. A one-way analysis of variance (ANOVA) followed by Tukey's post hoc analysis was used to compare gene expression between the three groups. P values less than 0.05 were considered to be statistically significant. All statistical analyses were performed on a personal computer using the Stat View version 5.0 statistical software package (Abacus Concepts, Berkeley, CA).
Six miRNAs up-regulated in IL-17 producing T cells in microarray analysis
Six miRNAs up-regulated in IL-17 producing T cells in microarray analysis.
6 miRNAs up-regulated
RA patient PBMCs exhibit increased expression of Let-7a, miR-26a, 146a,b, 150, and 155
The synovium from RA patients exhibit increased expression of miR-146a,b, 150, and 155
MiR-146a expressed in IL-17 producing cells
MiR-146a was previuosly reported to be expressed in the T cells in the RA synovium . We therefore confirmed whether miR-146a expressed in the IL-17 producing T cells using the double staining of in situ hybridization and immunohistochemistry of IL-17. MiR-146a expression was mainly observed in the superficial and sublining layers as previuosly described . In addition, IL-17 was seen in the RA synovium, while it was not seed in OA synovium. Double staining revealed that miR-146 + cells merged IL-17, thus indicating miR-146a to be expressed in IL-17 producing T cells (Figure 4B).
Recently, a potential link between miRNA and several human diseases has been revealed. For example, the expression of let-7 has been shown to be lower in lung cancer tissue than in normal lung tissue, and such down-regulation may promote high levels of expression of the Ras gene . In addition, the expression of miR-143 and miR-145 is reduced in colon cancer tissue. Evidence of miRNA function in conditions such as leukemia, viral infection, and DiGeorge syndrome has been reported [31, 32]. Recent reports have suggested that several miRNA might participate in the pathogenesis of RA [21, 24–26].
miRNAs have been investigated because of their potential clinical application for therapeutic methods. Therapeutic trials aimed at silencing miRNA in vivo have been conducted [32, 33]. Tazawa et al. demonstrated that miR-34a is down-regulated in human colon cancer, and that tumor growth in mice is significantly inhibited by the local injection of synthetic double stranded miR-34a in vivo. The intra-articular injection of double stranded miR-15a successfully induced cell apoptosis by inhibiting the translation of BCL2 protein in the synovium in arthritic mice. This suggested that the intra-articular injection of synthetic miRNAs can regulate the endogenous miRNAs in arthritic synovia . The discovery of a new lineage of CD4+ effecter T helper (Th) cells that selectively produce IL-17 in mice has provided exciting new insights into immune regulation, host defense, and the pathogenesis of autoimmune and other chronic inflammatory disorders. Although IL-17 plays a crucial role in RA pathogenesis, the differentiation mechanism of human Th17 cells from CD4+ T cells is unclear. MiRNAs are reported to be a important factor in the differentiation of T cells . Therefore, identification of miRNAs in Th17 cell differentiation could elucidate a new mechanism of RA pathogenesis, subsequently lead a novel therapeutic approach to the RA. In the current study, IL-17 producing T cells were expanded from PBMCs from healthy volunteers and the expression pattern of miRNAs in the differentiation of IL-17 producing cells was analyzed by using a Microarray analysis. This demonstrated that let-7a, miR-26a, 146a,b, 150, and 155 were significantly upregulated in the differentiation of IL-17 producing cells. There are several reports about Foxp3, RORγt, IRAK1 and SOCS1in the differentiation of IL-17 producing T cells. Foxp3 is the transcription factor controlling regulatory T cell development, while, RORγt is the master regulator that directs the differentiation program of Th17 cells . The expression level of RORγt in expanded IL-17 producing T cells was significantly higher than in non-expanded cells, while Foxp3 was expressed more strongly in non-expanded cells in comparison to expanded cells, indicating that the IL-17 producing T cells were expanded. IRAK1 and SOCS1 not only play a crucial role in Th17 cell differentiation, but also are regulated by miR-146a/b and miR-155 respectively [39, 40]. In the current series, the expression level of SOCS1 in expanded IL-17 producing T cells was significantly higher than in non-expanded cells. There was no significant difference in the expression level of IRAK1 between the expanded and non expanded cells at the mRNA level. However IRAK1 protein was decreased in the expanded cells, which suggested miR-146a/b might inhibit the translation to IRAK1 protein from mRNA.
MiR-146a was expressed intensely with IL-17 expression in PBMC from the patients with early stage of RA and high disease activity. miR-146a/b was associated with high disease activity, and miR-150 was intensely expressed in the patients with severe joint destruction. MiR-150 has not been associated with the high expression of miR-146 and miR-155 in PBMC and synovium of RA in previous reports [24–26]. MiR-150 might also play a role in RA pathogenesis. The expression of miR-146a and IL-17 was high in PBMC in patients with low score of Larsen grade and short disease duration in the current series. A patient with a high score of Larsen grade and long disease duration showed a low expression level of miR-146a, while that of miR-150 was high. Therefore, the high expression of miR-146a in PBMC was strongly associated with IL-17 expression, especially at the early stage of RA. Severe joint destruction with high disease activity was associated with the expression of miR-150 in the synovium. The number of patients in the current study was small, therefore, further examination is necessary to clarify the relationship between these miRNAs and disease activity, including the expression level of IL-17. Our results revealed miR-146a to be expressed in the IL-17 producing T cells. In a previous report, accumulated CD3+ T cells were observed to express miR-146, thereby suggesting that miR-146 might play a role in the persistent inflammation in RA via a T cell network, which strongly supports that miR-146a thus expressed in Th17 cells .
These results indicated that miR-146a/b, miR-150 and miR-155 were associated with IL-17 expression in the PBMC and synovium in the RA patients, especially expression of miR-146a in IL-17 producing cells was comfirmed. Their altered expression in the stage and activity of RA suggested that they might play a role in the pathogenesis of RA via IL-17 expression. In addition, our results should be developed the excessive analysis of the function of these miRNAs in the RA to lead the novel treatment.
erythrocyte sediment rate
nonsteroidal anti-inflammatory drugs
retinoid-related orphan receptor γ
suppressor of cytokine signaling
IL-1 receptor-associated kinase 1.
The authors thank Professor Michihiro Hide for permission to use the instruments and Ms Kaori Ishii for technical support with auto MACS. (Department of Dermatology, Graduate School of Biomedical Sciences, Hiroshima University, Japan).
- Gardner DL: In: Pathological basis of the connective tissue diseases. London:Edward Arnold. 1992, 444-526. Rheumatoid arthritis: cell and tissue pathologyGoogle Scholar
- Harrington LE, Hatton RD, Mangan PR, Turner H, Murphy TL, Murphy KM, Weaver CT: Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol. 2005, 6: 1123-1132. 10.1038/ni1254.View ArticlePubMedGoogle Scholar
- Park H, Li Z, Yang XO, Chang SH, Nurieva R, Wang YH, Wang Y, Hood L, Zhu Z, Tian Q, et al.: A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol. 2005, 6: 1133-1141. 10.1038/ni1261.View ArticlePubMedPubMed CentralGoogle Scholar
- Spriggs MK: Interleukin-17 and its receptor. J Clin Immunol. 1997, 17: 366-369. 10.1023/A:1027360106635.View ArticlePubMedGoogle Scholar
- Acosta-Rodriguez EV, Napoletani G, Lanzavecchia A, Sallusto F: Interleukins 1β and 6 but not transforming growth factor-β are essential for the differentiation of interleukin 17-producing human T helper cells. Nat Immunol. 2007, 8: 942-949. 10.1038/ni1496.View ArticlePubMedGoogle Scholar
- Chen Z, Tato CM, Muul L, Laurence A, O'Shea JJ: Distinct regulation of interleukin-17 in human T helper lymphocytes. Arthritis Rheum. 2007, 56: 2936-2946. 10.1002/art.22866.View ArticlePubMedPubMed CentralGoogle Scholar
- Wilson NJ, Boniface K, Chan JR, McKenzie B, Blumenschein WM, Mattson JD, Basham B, Smith K, Chen T, Morel F, et al.: Development, cytokine profile and function of human interleukin 17-producing helper T cells. Nat Immunol. 2007, 8: 950-957. 10.1038/ni1497.View ArticlePubMedGoogle Scholar
- van Beelen AJ, Zelinkova Z, Taanman-Kueter EW, Muller FJ, Hommes DW, Zaat SAJ, Kapsenberg ML, de Jong EC: Stimulation of the intracellular bacterial sensor NOD2 programs dendritic cells to promote interleukin-17 production in human memory T cells. Immunity. 2007, 27: 1-10. 10.1016/j.immuni.2007.07.003.View ArticleGoogle Scholar
- Sato K, Takayanagi H: Osteoclasts, rheumatoid arthritis, and osteoimmunology. Curr Opin Rheumatol. 2006, 18: 419-426. 10.1097/01.bor.0000231912.24740.a5.View ArticlePubMedGoogle Scholar
- Takayanagi H: Osteoimmunology: shared mechanisms and crosstalk between the immune and bone systems. Nat Rev Immunol. 2007, 7: 292-304. 10.1038/nri2062.View ArticlePubMedGoogle Scholar
- Kojiro S: Th17 cells and rheumatoid arthritis-from the standpoint of osteoclast differentiation. Allergology International. 2008, 57: 109-114. 10.2332/allergolint.R-07-158.View ArticleGoogle Scholar
- Chabaud M, Durand JM, Buchs N, Fossiez F, Page G, Frappart L, Miossec P: Human interleukin-17: a T cell-derived proinflammatory cytokine produced by the rheumatoid synovium. Arthritis Rheum. 1999, 42: 963-970. 10.1002/1529-0131(199905)42:5<963::AID-ANR15>3.0.CO;2-E.View ArticlePubMedGoogle Scholar
- Kotake S, Udagawa N, Takahashi N, Matsuzaki K, Itoh K, Ishiyama S, Saito S, Inoue K, Kamatani N, Matthew T: IL-17 in synovial fluids from patients with rheumatoid arthritis is a potent stimulator of osteoclastogenesis. J Clin Invest. 1999, 103: 1345-1352. 10.1172/JCI5703.View ArticlePubMedPubMed CentralGoogle Scholar
- Ziolkowska M, Koc A, Luszczykiewics G, Ksiezopolska-Pietrzak K, Klimczak E, Chwalinska-Sadowska H, Malinski W: High levels of IL-17 in rheumatoid arthritis patients: IL-15 triggers in vitro IL-17production via cyclosporin A-sensitive mechanism. J Immunol. 2000, 164: 2832-2838.View ArticlePubMedGoogle Scholar
- Ambros V: The functions of animal microRNAs. Nature. 2004, 431: 350-355. 10.1038/nature02871.View ArticlePubMedGoogle Scholar
- Bartel DP, Chen CZ: MicroRNAs: genomics, biogenesis, mechanism, andfunction. Cell. 2004, 116: 281-297. 10.1016/S0092-8674(04)00045-5.View ArticlePubMedGoogle Scholar
- Farh KK, Grimson A, Jan C, Lewis BP, Johnson WK, Lim LP, Burge CB, Bartel DP: The widespread impact of mammalian microRNAs on mRNArepression and evolution. Science. 2005, 310: 1817-1821. 10.1126/science.1121158.View ArticlePubMedGoogle Scholar
- Denli AM, Tops BB, Plasterk RH, Ketting RF, Hannon GJ: Processing of primary microRNAs by the Microprocessor complex. Nature. 2004, 432: 231-235. 10.1038/nature03049.View ArticlePubMedGoogle Scholar
- Gregory RI, Yan KP, Amuthan G, Chendrimada T, Doratotaj B, Cooch N, Shiekhattar R: The Microprocessor complex mediates the genesis of microRNAs. Nature. 2004, 432: 235-240. 10.1038/nature03120.View ArticlePubMedGoogle Scholar
- Lee Y, Ahn C, Han J, Choi H, Kim J, Yim J, Lee J, Provost P, Redmark O, Kim S, et al.: The nuclear RNase III Drosha initiates microRNA processing. Nature. 2003, 425: 415-419. 10.1038/nature01957.View ArticlePubMedGoogle Scholar
- Chendrimada TP, Gregory RI, Kumaraswamy E, Norman J, Cooch N, Nishikura K, Shiekhattar R: TRBP recruits the Dicer complex toAgo2 for microRNA processing and gene silencing. Nature. 2005, 436: 740-744. 10.1038/nature03868.View ArticlePubMedPubMed CentralGoogle Scholar
- Calin GA, Sevignani C, Dumitru CD, Hyslop T, Noch E, Yendamuri S, Shimizu M, Rattan S, Bullrich F, Negrini M: Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA. 2004, 101: 2999-3004. 10.1073/pnas.0307323101.View ArticlePubMedPubMed CentralGoogle Scholar
- Lecellier CH, Dunoyer P, Arar K, Lehmann-Che J, Eyquem S, Himber C, Saib A, Voinnet O: A cellular microRNA mediates antiviral defense in human cells. Science. 2005, 308: 557-560. 10.1126/science.1108784.View ArticlePubMedGoogle Scholar
- Stanczyk J, Pedrioli DM, Brentano F, Sanchez-Pernaute O, Kolling C, Gay RE, Detmar M, Steffen Gay, Kyburz D: Altered expression of microRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis. Arthritis Rheum. 2008, 58: 1001-1009. 10.1002/art.23386.View ArticlePubMedGoogle Scholar
- Nakasa T, Miyaki S, Okubo A, Hashimoto M, Nishida K, Ochi M: Expression of microRNA-146 in rheumatoid arthritis synovial tissue. Arthritis Rheum. 2008, 58: 1284-1292. 10.1002/art.23429.View ArticlePubMedPubMed CentralGoogle Scholar
- Pauley KM, Satoh M, Chan AL, Bubb MR, Reeves WH, Chan EK: Upregulated miR-146a expression in peripheral blood mononuclear cells from rheumatoid arthritis patients. Arthritis Res Ther. 2008, 10: R101-10.1186/ar2344.View ArticlePubMedPubMed CentralGoogle Scholar
- Changsheng D, Chang L, Jiuhong K, Guixian Z, Zhiqiang Y, Shichao H, Zhenxin L, Zhiiying W, Gang P: MicroRNA-326 regulates Th-17 differentiation and associated with the pathogenesis of multiple sclerosis. Nat Immunol. 2009, 10: 1252-1259. 10.1038/ni.1798.View ArticleGoogle Scholar
- Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS: The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988, 31: 315-324. 10.1002/art.1780310302.View ArticlePubMedGoogle Scholar
- Amadi-obi A, Yu CR, Liu X, Mahdi RM, Clarke GL, Nussenblatt RB, Gery L, Lee YS, Egwuagu CE: Th17 cells contribute to uveits and scleritis and are expanded by IL-2 and inhibited by IL-27/STAT1. Nat Med. 2007, 13: 711-718. 10.1038/nm1585.View ArticlePubMedGoogle Scholar
- Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Emmanuel L, Reinert KL, Brown D, Slack FJ: RAS is regulated by the let-7 microRNA family. Cell. 2005, 120: 635-647. 10.1016/j.cell.2005.01.014.View ArticlePubMedGoogle Scholar
- Michael MZ, O'Conner SM, van Holst Pellekaan NG, Young GP, James RJ: Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol Cancer Res. 2003, 1: 882-891.PubMedGoogle Scholar
- Krutzfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M: Silencing of microRNAs in vivo with 'antagomirs'. Nature. 2005, 438: 685-689. 10.1038/nature04303.View ArticlePubMedGoogle Scholar
- Yang B, Lin H, Xiao J, Lu Y, Luo X, Li B, Zhang Y, Xu C, Bai Y, Wang H: The musclespecific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. Nat Med. 2007, 13: 486-491. 10.1038/nm1569.View ArticlePubMedGoogle Scholar
- Tazawa H, Tsuchiya N, Izumiya M, Nakagama H: Tumor-suppressive miR-34a induces senescence-like growth arrest through modulation of the E2F pathway in human colon cancer cells. Proc Natl Acad Sci USA. 2007, 104: 15472-15477. 10.1073/pnas.0707351104.View ArticlePubMedPubMed CentralGoogle Scholar
- Nagata Y, Nakasa T, Mochizuki Y, Ishikawa M, Miyaki S, Shibuya H, Yamasaki K, Adachi N, Asahara H, Mitsuo Ochi: Induction of apoptosis in the synovium of mice with autoantibody-mediated arthritis by the intraarticular injection of double-stranded MicroRNA-15a. Arthritis Rheum. 2009, 60: 2677-2683. 10.1002/art.24762.View ArticlePubMedGoogle Scholar
- Turner M, Vigorito E: Regulation of B- and T-cell differentiation by asingle microRNA. Biochem Soc Trans. 2008, 36: 532-533. 10.1042/BST0360531.View ArticleGoogle Scholar
- Hori S, Nomura T, Sakaguchi S: Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003, 299: 1057-1061. 10.1126/science.1079490.View ArticlePubMedGoogle Scholar
- Ivanov II, Mackenzie BS, Zhou L, Tadokoro CE, Lepelley A, Lafaille JJ, Cua DJ, Littman DR: The orphan nuclear receptor RORγt directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell. 2006, 126: 1121-1131. 10.1016/j.cell.2006.07.035.View ArticlePubMedGoogle Scholar
- Yoshimura A, Naka T, Kubo M: SOCS proteins,cytokine signaling and immune regulation. Nat Reu Immunol. 2007, 7 (6): 454-465. 10.1038/nri2093.View ArticleGoogle Scholar
- Li-Fan L, To-Ha T, Dinis PC, Ashutosh C, Kubo M, Tanaka K, Loeb GB, Lee H, Yoshimura A, Rajewsky K, et al.: Foxp3-Dependent MicroRNA155 Confers Competitive Fitness to Regulatory T Cells by Targeting SOCS1 Protein. Immunity. 2009, 30: 80-91. 10.1016/j.immuni.2008.11.010.View ArticleGoogle Scholar
- Slack FJ, Basson M, Liu Z, Ambros V, Horvitz HR, Ruvkun G: The lin-41 RBCC gene acts in the C. elegans heterochronic pathway between the let-7 regulatory RNA and the LIN-29 transcription factor. Mol Cell. 2000, 5: 659-669. 10.1016/S1097-2765(00)80245-2.View ArticlePubMedGoogle Scholar
- Abrahante JE, Daul AL, Li M, Volk ML, Tennessen JM, Miller EA, Rougvie AE: The Caenorhabditis elegans hunchback-like gene lin-57/hbl-1 controls developmental time and is regulated by microRNAs. Dev Cell. 2003, 4: 625-637. 10.1016/S1534-5807(03)00127-8.View ArticlePubMedGoogle Scholar
- Lin SY, Johnson SM, Abraham M, Vella MC, Pasquinelli A, Gamberi C, Gottlieb E, Slack FJ: The C. elegans hunchback homolog, hbl-1, controls temporal patterning and is a probable microRNA target. Dev Cell. 2003, 4: 639-650. 10.1016/S1534-5807(03)00124-2.View ArticlePubMedGoogle Scholar
- You-Chin L, Li-Ching H, Ming-Wei K, John Y, Huan-Hsien K, Wan-Lin L, Ruey-Jen L, Alice LY, Wen-Hsiung L: Human TRIM71 and Its Nematode Homologue Are Targets of let-7 MicroRNA and Its Zebrafish Orthologue Is Essential for Development. Mol Biol Evol. 2007, 24 (11): 2525-2534. 10.1093/molbev/msm195.View ArticleGoogle Scholar
- Chung FW, Ross LT: MicroRNA-26a Targets the Histone Methyltransferase Enhancer of Zeste homolog 2 during Myogenesis. J Biol chem. 2008, 283 (15): 9836-9843. 10.1074/jbc.M705871200.View ArticleGoogle Scholar
- Junaith SM, Michael AL, Aladin MB: Mechanical Stretch Upregulates MicroRNA-26a and Induces Human Airway Smooth Muscle Hypertrophy by Suppressing Glycogen Synthase Kinase-3β. J Biol chem. 2010,Google Scholar
- Taganov KD, Boldin MP, Chang KJ, Baltimore D: NFkappaB dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci USA. 2008, 103: 12481-12486. 10.1073/pnas.0605298103.View ArticleGoogle Scholar
- Hou J, Wang P, Lin L, Liu X, Ma F, An H, Wang Z, Cao X: MicroRNA-146a feedback inhibits RIG-I-dependent type I IFN production in macrophages by targeting TRAF6, IRAK1, and IRAK2. J Immunol. 2009, 183: 2150-2158. 10.4049/jimmunol.0900707.View ArticlePubMedGoogle Scholar
- Barroga Charlene F, Pham Hang, Kaushansky Kenneth: Thrombopoietin regulates c-Myb expression by modulating micro RNA 150 expression. Experimental Hematology. 2008, 36: 1585-1592. 10.1016/j.exphem.2008.07.001.View ArticlePubMedPubMed CentralGoogle Scholar
- Tili E, Michaille JJ, Cimino A, Costinean S, Dumitru CD, Adair B, Fabbri M, Alder H, Liu CG, Calin GA, et al.: Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-alpha stimulation and their possible roles in regulating the response to endotoxin shock. J Immunol. 2007, 179: 5082-5089.View ArticlePubMedGoogle Scholar
- Ceppi M, Pereira PM, Dunand-Sauthier I, Barras E, Reith W, Santos MA, Pierre P: MicroRNA-155 modulates the interleukin-1 signaling pathway in activated human monocyte- derived dendritic cells. Proc Natl Acad Sci USA. 2009, 106: 2735-2740. 10.1073/pnas.0811073106.View ArticlePubMedPubMed CentralGoogle Scholar
- Martinez-Nunez RT, Louafi F, Friedmann PS, Sanchez-Elsner T: MicroRNA-155 modulates the pathogen binding ability of dendritic cells (DCs) by down-regulation of DC-specific intercellular adhesion molecule-3 grabbing non-integrin (DCSIGN). J Biol Chem. 2009, 284: 16334-16342. 10.1074/jbc.M109.011601.View ArticlePubMedPubMed CentralGoogle Scholar
- Teng G, Hakimpour P, Landgraf P, Rice A, Tuschl T, Casellas R, Papavasiliou FN: MicroRNA-155 is a negative regulator of activation-induced cytidine deaminase. Immunity. 2008, 28: 621-629. 10.1016/j.immuni.2008.03.015.View ArticlePubMedPubMed CentralGoogle Scholar
- The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2474/11/209/prepub
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.