Diagnostic Performances of Anti-Cyclic Citrullinated Peptides Antibody and Antifilaggrin Antibody in Korean Patients with Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a systemic autoimmune disease of unknown etiology. We studied the diagnostic performances of anti-cyclic citrullinated peptides antibody (anti-CCP) assay and recombinant anti-citrullinated filaggrin antibody (AFA) assay by enzyme linked immunosorbent assay (ELISA) in patients with RA in Korea. Diagnostic performances of the anti-CCP assay and AFA assay were compared with that of rheumatoid factor (RF) latex fixation test. RF, anti-CCP, and AFA assays were performed in 324 RA patients, 251 control patients, and 286 healthy subjects. The optimal cut off values of each assay were determined at the maximal point of area under the curve by receiver-operator characteristics (ROC) curve. Sensitivity (72.8%) and specificity (92.0%) of anti-CCP were better than those of AFA (70.3%, 70.5%), respectively. The diagnostic performance of RF showed a sensitivity of 80.6% and a specificity of 78.5%. Anti-CCP and AFA showed positivity in 23.8% and 17.3% of seronegative RA patients, respectively. In conclusion, we consider that anti-CCP could be very useful serological assay for the diagnosis of RA, because anti-CCP revealed higher diagnostic specificity than RF and AFA at the optimal cut off values and could be performed by easy, convenient ELISA method

Autoimmune Responses in the Rheumatoid Synovium

Rene Toes and Tom Huizinga discuss a new study indicating that lymphoneogenesis in the inflamed synovial tissue of patients with rheumatoid arthritis is fostering potentially pathogenic immune responses

Double walled carbon nanotubes promote the overproduction of extracellular protein-like polymers in Nitzschia palea: An adhesive response for an adaptive issue

The present study assessed the effects of double-wall carbon nanotubes (DWCNTs) dispersed in the presence of a realistic concentration of natural organic matter (NOM, 10 mg L−1) on the benthic diatom Nitzschia palea using toxicity tests and quantitative/qualitative extracellular polymeric substances (EPS) assays. No toxic effect was observed. A growth delay was measured after 48 h of exposure to concentrations of DWCNTs ranging from 1 mg L−1 (∼29%) to 50 mg L−1 (∼84%). Extracellular carbohydrates and proteins were extracted using a sequential multi-methods protocol to collect soluble, hydrophobic and ion-bridged extracellular polymeric substances (EPS). Extracted EPS were analyzed by colorimetric assays and size exclusion chromatography. The results highlighted a higher EPS concentration in exposed cultures that was primarily caused by an overproduction of protein-like polymers (protein or glycoproteins, PLPs). Such EPS overproduction and increase in proteins/carbohydrates ratio can partially explain the observed growth inhibition. EPS were preferentially extracted using hydrophobic conditions and were mainly composed of PLPs with either low (10 kDa) or high (174 kDa) molecular weights. These data highlights the affinity between DWCNTs and EPS, which is primarily driven by both physical and hydrophobic interactions. This indicates that N. palea can respond to DWCNTs by forming an EPS network optimized for adhering to and efficiently wrapping DWCNTs

The response of extracellular polymeric substances production by phototrophic biofilms to a sequential disturbance strongly depends on environmental conditions

Phototrophic biofilms are exposed to multiple stressors that can affect them both directly and indirectly. By modifying either the composition of the community or the physiology of the microorganisms, press stressors may indirectly impact the ability of the biofilms to cope with disturbances. Extracellular polymeric substances (EPS) produced by the biofilm are known to play an important role in its resilience to various stresses. The aim of this study was to decipher to what extent slight modifications of environmental conditions could alter the resilience of phototrophic biofilm EPS to a realistic sequential disturbance (4-day copper exposure followed by a 14-day dry period). By using very simplified biofilms with a single algal strain, we focused solely on physiological effects. The biofilms, composed by the non-axenic strains of a green alga (Uronema confervicolum) or a diatom (Nitzschia palea) were grown in artificial channels in six different conditions of light intensity, temperature and phosphorous concentration. EPS quantity (total organic carbon) and quality (ratio protein/polysaccharide, PN/PS) were measured before and at the end of the disturbance, and after a 14-day rewetting period. The diatom biofilm accumulated more biomass at the highest temperature, with lower EPS content and lower PN/PS ratio while green alga biofilm accumulated more biomass at the highest light condition with lower EPS content and lower PN/PS ratio. Temperature, light intensity, and P concentration significantly modified the resistance and/or recovery of EPS quality and quantity, differently for the two biofilms. An increase in light intensity, which had effect neither on the diatom biofilm growth nor on EPS production before disturbance, increased the resistance of EPS quantity and the resilience of EPS quality. These results emphasize the importance of considering the modulation of community resilience ability by environmental conditions, which remains scarce in the literature

Structure and pathogenicity of antibodies specific for citrullinated collagen type II in experimental arthritis

Antibodies to citrulline-modifi ed proteins have a high diagnostic value in rheumatoid arthritis (RA). However, their biological role in disease development is still unclear. To obtain insight into this question, a panel of mouse monoclonal antibodies was generated against a major triple helical collagen type II (CII) epitope (position 359 – 369; ARGLTGRPGDA) with or without arginines modifi ed by citrullination. These antibodies bind cartilage and synovial tissue, and mediate arthritis in mice. Detection of citrullinated CII from RA patients ’ synovial fl uid demonstrates that cartilage-derived CII is indeed citrullinated in vivo. The structure determination of a Fab fragment of one of these antibodies in complex with a citrullinated peptide showed a surprising beta -turn conformation of the peptide and provided information on citrulline recognition. Based on these findings, we propose that autoimmunity to CII, leading to the production of antibodies specific for both native and citrullinated CII, is an important pathogenic factor in the development of RA

Autoantibodies to BRAF, a new family of autoantibodies associated with rheumatoid arthritis

International audienceBRAF (v raf murine sarcoma viral oncogene homologue B1) is a serine-threonine kinase involved in the mitogen-activated protein kinase (MAPK) signalling pathway, known to be implicated in the production of pro-inflammatory cytokines.We have observed that sera from rheumatoid arthritis (RA) patients recognize the BRAF's catalytic domain, which encompasses amino acids 416 to 766. Here, we identify peptide targets of anti-BRAF autoantibodies and test whether anti-BRAF autoantibodies may interfere with BRAF kinase activity.METHODS:Anti-BRAF autoantibodies were detected by ELISA (enzyme-linked immunosorbent assay) in the serum of RA patients and controls, using 40 overlapping 20mer peptides encompassing the catalytic domain of BRAF as immunosorbents. To test whether autoantibodies to BRAF influence BRAF kinase activity, we developed an in vitro phosphorylation assay of MEK1 (mitogen extracellular regulated kinase), a major BRAF substrate. MEK1 phosphorylation by BRAF was tested in the presence of purified anti-BRAF autoantibodies from RA patients or control antibody.RESULTS:We found that one BRAF peptide, P25 (656 to 675), is specifically recognized by autoantibodies from RA patients. Of interest, anti-P25 autoantibodies are detected in 21% of anti-CCP (cyclic citrullinated peptides) negative RA patients. Anti-BRAF autoantibodies activate the in vitro phosphorylation of MEK1 mediated by BRAF.CONCLUSIONS:Anti-BRAF autoantibodies from RA patients preferentially recognize one BRAF peptide: P25. Autoantibody responses to P25 are detected in 21% of anti-CCP negative RA patients. Most anti-BRAF autoantibodies activate BRAF kinase activity

Ectopic Lymphoid Structures Support Ongoing Production of Class-Switched Autoantibodies in Rheumatoid Synovium

Costantino Pitzalis and colleagues show that lymphoid structures in synovial tissue of patients with rheumatoid arthritis support production of anti-citrullinated peptide antibodies, which continues following transplantation into SCID mice

Nitration of the Pollen Allergen Bet v 1.0101 Enhances the Presentation of Bet v 1-Derived Peptides by HLA-DR on Human Dendritic Cells

Nitration of pollen derived allergens can occur by NO2 and ozone in polluted air and it has already been shown that nitrated major birch (Betula verrucosa) pollen allergen Bet v 1.0101 (Bet v 1) exhibits an increased potency to trigger an immune response. However, the mechanisms by which nitration might contribute to the induction of allergy are still unknown. In this study, we assessed the effect of chemically induced nitration of Bet v 1 on the generation of HLA-DR associated peptides. Human dendritic cells were loaded with unmodified Bet v 1 or nitrated Bet v 1, and the naturally processed HLA-DR associated peptides were subsequently identified by liquid chromatography-mass spectrometry. Nitration of Bet v 1 resulted in enhanced presentation of allergen-derived HLA-DR-associated peptides. Both the copy number of Bet v 1 derived peptides as well as the number of nested clusters was increased. Our study shows that nitration of Bet v 1 alters antigen processing and presentation via HLA-DR, by enhancing both the quality and the quantity of the Bet v 1-specific peptide repertoire. These findings indicate that air pollution can contribute to allergic diseases and might also shed light on the analogous events concerning the nitration of self-proteins