ABSTRACT
Objective ① To Screen for the miRNAs differently expressed in the peripheral blood mono-nuclear cells (PBMCs) of rheumatoid arthritis (RA) by microarray experiments.② To further evaluate the expression of miR-155 in PBMCs of RA.③ To determine the relevance between the expression of miR-155 and clinical as well as laboratory features.④ To test whether inflammatory mediators can induce miR-155 expression in PBMCs of RA.Methods ① Total RNA was isolated from peripheral blood mononuclear cells obtained from 5 patients of RA and 5 normal controls.Expression profiling of miRNAs was performed in a microarray analysis.② MiR-155 was identified for further study by stem-loop real-time RT-PCR based on SYBR-Green.PBMC from 26 patients of RA and 23 normal controls were collected.③ Association between miR-155 and the clinical and laboratory features of RA was evaluated.④Induction of miR-155 following stimulation with TNF-α, IFN-γ and LPS of cultures of RA PBMCs was examined by real-time RT-PCR.Statistical analysis was done with student's t test, paired t test, and ANOVA, Spearman correlation.Results ① Expression profiling of miRNAs revealed significant differential expression of 14 miRNAs, of which signal intensity changed over two times.MiR-155 was up-regulated in PBMCs of RA than in normal controls (t=9.218, P=0.001).② The expression level of miR-155 had a positive correlation with serum CRP level (r=0.57, P=0.002).③ Expression of miR-155 was markedly up-regulated in PBMCs of RA after stimulated with TNF-α,IFN-γ and LPS, especially with TNF-α.Conclusions The expression of miR-155 is induced by stimulating with TNF-α, IFN-γ and LPS.MiR-155 may be a regulator in RA pathogenesis.Further studies are required to elucidate the function of miR-155.
ABSTRACT
Objective To detect antibodies against human papilloma virus-47 E2m345-362 peptide whichis homologous to profilaggri306324 peptide and anti-citrullinated human papilloma virus-47 E2345-362 peptide antibodies in rheumatoid arthritis(RA)and to investigate its role in the pathogenesis of RA.Methods Serum samples were obtained from 119 patients with RA, other rheumatic diseases and healthy individuals.We searched the homologus sequence of profilaggrin306-324peptide by using NCBI (the National Center for Bioteehnology Information)BLAST (basic local alignment search tool),and synthesized human papilloma virus-47 E2345-362 peptide which was highly homologous to profilaggrin306-324 peptide and the citrullinated Human papilloma virus-47 E2345-362 peptide.The presence of antibodies against E2345-362 peptide and citrullinated E2345-362 peptide was examined by enzyme-linked immunosorbent assay(ELISA).The associations between these antibodies and the clinical features of RA were evaluated.Results ①(41.2%)and titer (AU was 105.7)of anti citrullinated E2345-362 peptide antibodies in RA were significantly higher than those of patients with other rheumatic diseases and healthy individuals.However,the prevalence of anti-E2345-362 peptide antibodies in RA patients was similar to that of patients with other rheumatic diseases and healthy individuals(P>0.05).②The samples that were pre-incubated with cyclic citrullinated peptide (CCP) had lower titer of anti-citrulllinated E2345-362 peptide antibodies.③The titers of anti-CCP antibodies and anti-PAD14 antibodies in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negtive patients.It showed significant correlations between anti-citrulllinated E2345-362 peptide antibodies and anti-PAD14 antibodies(r=0.485,P<0.01).④ DAS28 score,ESR,X-ray stages,AKA in anti-citrullinated E2345-362 positive patients were higher than those of anti-citrullinated E2345-362 negative patients.Conclusion The presence of anti-citrullinated E2345-362 peptide antibodies in RA indicates that HPV-47 E2 may act as an auto-antigen which may play an important role in the pathogenesis of RA.The increasing of PAD14 may make it easy for HPV-47 E2 to be citrullinated and may induce the subsequent auto-immune reactions.
ABSTRACT
Objective:To evaluate the effect of mucosal administration of altered collagen Ⅱ(CⅡ)263-272 peptide(267Q→A,270K→A and 271G→A) on collagen induced arthritis(CIA),and to explore the mechanism of the inhibitory effect of the altered CⅡ263-272 peptide on CIA.Methods:CIA was induced in Lewis rats by immunization with bovine CⅡ.Altered CⅡ263-272 peptide was given intranasally beginning from the onset of arthritis(100 ?g/dose,daily for 5 doses and continuing every other day for other 7 doses).Wild CⅡ263-272 peptide(100 ?g/dose) or PBS was administered as controls with the same procedure.Therapeutic effects were evaluated by arthritis scores,body weight change,and joint pathologic scores.The anti-CⅡ antibody and its subtypes were measured with ELISA.The cytokines of IFN-? and IL-10 were measured with ELISA.The induction of regulatory T cells was assessed by FACS analysis of percentage of peripheral CD4+CD25+ T cells,and by real-time PCR analysis of the expression of Foxp3 and TGF-? mRNA.Results:(1) Following treatment with the altered CⅡ263-272 peptide,arthiritis scores were reduced and body weight was increased.The mean arthritis scores of rats treated with altered peptide,wild peptide and PBS were 2.50?2.43,4.50?2.23 and 6.33?2.73,respectively.The altered peptide could retard the histologic lesion of the joints.(2) The titers of anti-CⅡ antibodies IgG and IgG1 in the three groups were similar,but the IgG2a in altered peptide-treated rats decreased markedly as compared with PBS-treated rats(0.56?0.19 vs 0.95?0.29,P