[Show abstract][Hide abstract] ABSTRACT: Intra-articular glucocorticoid treatment (IAGC) is widely used for symptom relief in arthritis. However, knowledge of factors predicting treatment outcome is limited. The aim of the present study was to identify response predictors of IAGC for knee synovitis in patients with rheumatoid arthritis (RA).
Arthritis research & therapy. 06/2014; 16(3):R129.
[Show abstract][Hide abstract] ABSTRACT: Immunological events in the lungs might trigger production of anti-citrullinated protein antibodies during early rheumatoid arthritis (RA). We investigated the presence of shared immunological citrullinated targets in joints and lungs of patients with RA.
Proteins extracted from bronchial (n=6) and synovial (n=7) biopsy specimens from patients with RA were investigated by mass spectrometry-based proteomics. One candidate peptide was synthesised and used to investigate by ELISA the presence of antibodies in patients with RA (n=393), healthy controls (n=152) and disease controls (n=236). HLA-DRB1 shared epitope (SE) alleles were detected in patients with RA.
Ten citrullinated peptides belonging to seven proteins were identified, with two peptides shared between the synovial and bronchial biopsy samples. Further analysis, using accurate mass and retention time, enabled detection of eight citrullinated peptides in synovial and seven in bronchial biopsy specimens, with five peptides shared between the synovial and bronchial biopsy specimens. Two citrullinated vimentin (cit-vim) peptides were detected in the majority of synovial and lung tissues. Antibodies to a synthesised cit-vim peptide candidate (covering both cit-vim peptides identified in vivo) were present in 1.8% of healthy controls, 15% of patients with RA, and 3.4% of disease controls. Antibodies to cit-vim peptide were associated with the presence of the SE alleles in RA.
Identical citrullinated peptides are present in bronchial and synovial tissues, which may be used as immunological targets for antibodies of patients with RA. The data provide further support for a link between lungs and joints in RA and identify potential targets for immunity that may mediate this link.
Annals of the rheumatic diseases 05/2014; · 8.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Smoking can induce autoantibodies in persons who are genetically predisposed to rheumatoid arthritis. We investigated the association between smoking and antiphospholipid antibodies (aPL) in systemic lupus erythematosus (SLE), a question not previously addressed. Further, we explored the relationship between smoking, aPL and vascular events (arterial and venous, VE).
In this cross-sectional study, clinical evaluation and questionnaire data were collected from 367 prevalent SLE patients. At the same time, we measured aPL (anticardiolipin (aCL), anti-β2 glycoprotein-1 (aβ2GP1) antibodies IgG/IgM/IgA, and lupus anticoagulant (LA)), and a large set of other SLE-associated autoantibodies for comparison. Association analyses using logistic regression models with smoking, (ever, former and current with never as reference) and antibody status as outcome variable were performed. As a secondary outcome, we investigated the associations between aPL, smoking and VE.
In multivariable-adjusted models ever, and in particular former, cigarette smoking was associated with the most pathogenic aPL; LA, aCL IgG and aβ2GP1 IgG. Other SLE-associated autoantibodies were not associated with smoking. The combination of smoking and aPL was strongly associated with VE. We noted a positive interaction between smoking-LA and smoking-'triple aPL' positivity for previous VE.
We investigated a large set of commonly occurring autoantibodies in SLE, but only aPL were positively associated with a history of smoking. This association was especially apparent in former smokers. Among ever regular smokers who were aPL positive, we observed a strikingly high frequency of former VE. The underlying mechanisms and temporality between smoking, aPL and VE need further investigations.
Annals of the rheumatic diseases 04/2014; · 8.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: RA patients with anti-collagen antibodies (anti-CII) are characterised by acute RA onset (Mullazehi et al, ARD 2007) and early joint erosions (Mullazehi et al ART 2012), as well as increased production of TNF by peripheral blood mononuclear cells from anti-CII immune complexes (IC) in vitro (Mullazehi et al A&R 2006). We have previously (abstract EWRR 2012) shown that polymorphonuclear neutrophil granulocytes (PMN) react towards anti-CII IC. The aim was to investigate whether functional PMN responses are associated with the clinical acute onset RA phenotype.
A set of 72 baseline patient sera (24 anti-CII positive, 24 anti-CII negative/RF positive and 24 anti-CII negative/RF negative) was chosen from a clinically well-characterised RA cohort with 2-year radiological follow-up (Larsen score). PMN and PBMC isolated from healthy donors were stimulated with anti-CII IC prepared with sera from the patients. PMN expression of CD16 and CD66b were measured by flow cytometry, and PMN production of myeloperoxidase (MPO) and IL-17, and PBMC production of TNF was measured with ELISA.
CD66b, MPO, and TNF were significantly up-regulated and CD16 was significantly down-regulated by IC made with anti-CII positive sera. Even anti-CII low positive sera were able to impact the expression of CD16, CD66b and TNF. There was linear correlation to CD16, CD66b, MPO and TNF production in relation to anti-CII levels (r = -0.3152, 0.6755, 0.2532 and 0.5846, respectively). CD16 correlated with early joint erosion (p = 0.024, 0.034, 0.046 at baseline, one and two years) and CD66b was associated with changes in joint erosion (p = 0.017 and 0.016, at one and two years compared to baseline, respectively). CD66b was associated with baseline CRP and PBMC production of TNF was associated with baseline ESR, in accordance to our earlier findings. No correlations were observed with IL-17.
We have earlier shown that PBMC anti-CII IC-induced production of TNF was associated with CRP and ESR in newly diagnosed RA patients. Here we show that PMN reactivity against anti-CII IC is uniquely associated with joint destruction. PMN reactivity towards anti-CII IC in cartilage of patients can contribute to joint destruction in newly diagnosed RA patients.
Annals of the rheumatic diseases 03/2014; 73 Suppl 1:A5. · 8.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate if changes in the lungs are present in rheumatoid arthritis (RA) patients early in the disease process and to address the contribution of these changes to the initiation of the disease.
24 RA patients with patient-reported symptom duration less than one year and naive to DMARD treatment and 15 healthy individuals were subjected to bronchoscopy and mucosal bronchial biopsies were retrieved. Histological analysis for identification of inducible bronchia associated lymphoid tissues (iBALT) and lymphocyte infiltration, as well as immunohistochemistry (IHC) for PAD enzymes, CD3, HLA-DQ and HLA-DR were performed. Presence of citrullinated targets were detected by IHC using biotinylated ACPA isolated from synovial fluid of RA patients. Mass spectrometry was used for identification of citrullinated epitopes in 6 lung and 8 synovial biopsies from RA patients. An in-house ELISA was setup to measure reactivity against new identified citrullinated targets in the serum of RA patients in two distinct early RA cohorts (n = 393) and a cohort of non-RA patients (n = 236) as disease controls.
Bronchial lymphocyte infiltration and iBALT formation was observed in 55% of the ACPA+ RA patients but only 17% of ACPA- patients and 7% of healthy volunteers. Higher expression of CD3, HLA-DQ, HLA-DR and citrullinated targets was observed in bronchial biopsies of ACPA+ as compare to ACPA- RA patients. BAL fluids were enriched in both IgG and IgA ACPA as compared to paired serum samples. Mass spectrometry identified 5 proteins in the synovium (in total 8 sites) and 4 in the lungs (in total 6 sites) containing citrullinated residues. Two vimentin derived citrullinated peptides were present in a majority of both synovial and lung biopsies with slightly higher citrullinated/unmodified peptides ratios in the smokers as compared to non-smokers.14.5% of the RA patients tested by ELISA showed antibody reactivity against the new identified citrullinated target compared to 3.4% in the disease controls.
Signs of inflammation and local ACPA enrichment are present early in bronchial tissues of ACPA+ RA patients. Shared citrullinated targets in the lung and joints as well as systemic reactivity against these targets are present in RA patients. Our findings support the notion that early inflammatory events in the lungs may represent a critical initiating factor in the development of ACPA+ RA.
Annals of the rheumatic diseases 03/2014; 73 Suppl 1:A4-5. · 8.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To analyse if predictors of radiographic progression differ between patients treated with or without prednisolone in early rheumatoid arthritis (RA). Radiographs of hands and feet were assessed using the modified Sharp/van der Heijde score and radiographic progression was defined as an increase in the total Sharp score above 5.8 (the smallest detectable change).
BMJ Open 01/2014; 4(7):e005246. · 1.58 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Selective immunoglobulin A deficiency is the most common primary immunodeficiency disorder that is strongly overrepresented among patients with celiac disease (CD). IgG antibodies against tissue transglutaminase (tTG) and deamidated gliadin peptides (DGP) serve as serological markers for CD in IgA deficient individuals, although the diagnostic value remains uncertain. The aim of this study was to investigate the prevalence of these markers in a large cohort of IgA deficient adults with confirmed or suspected CD and relate the findings to gluten free diet.
Sera from 488,156 individuals were screened for CD in seven Swedish clinical immunology laboratories between 1998 and 2012. In total, 356 out of 1,414 identified IgA deficient adults agreed to participate in this study and were resampled. Forty-seven IgA deficient blood donors served as controls. Analyses of IgG antibodies against tTG and DGP as well as HLA typing were performed and a questionnaire was used to investigate adherence to gluten free diet. Available biopsy results were collected.
Out of the 356 IgA deficient resampled adults, 67 (18.8%) were positive for IgG anti-tTG and 79 (22.2%) for IgG anti-DGP, 54 had biopsy confirmed CD. Among the 47 IgA deficient blood donors, 4 (9%) were positive for IgG anti-tTG and 8 (17%) for anti-DGP. Four were diagnosed with biopsy verified CD, however, 2 of the patients were negative for all markers. Sixty-eight of 69 individuals with positive IgG anti-tTG were HLA-DQ2/DQ8 positive whereas 7 (18.9%) of the 37 individuals positive for IgG anti-DGP alone were not.
IgG anti-tTG seems to be a more reliable marker for CD in IgA deficient adults whereas the diagnostic specificity of anti-DGP appears to be lower. High levels of IgG antibodies against tTG and DGP were frequently found in IgA deficient adults despite adhering to gluten free diet.
PLoS ONE 01/2014; 9(4):e93180. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective
The pathophysiology of the postpolio syndrome is not fully understood. Increased cytokine levels in cerebrospinal fluid and peripheral blood indicate a systemic inflammatory process. Decreased cytokine levels and the clinical effect of intravenous immunoglobulin treatment further indicate an inflammatory/immunological pathogenesis. The aim of the present study was to evaluate whether an autoimmune process follows the initial infection, by means of analyzing immune complexes.
Patients and methods
Circulating immune complexes were analyzed from blood samples of 20 postpolio patients and 95 healthy controls. To compensate for differences in age between patients and controls, a sub-analysis was performed using only the 30 oldest controls. Tumor necrosis factor-inducing properties of polyethylene glycol-precipitated immune complexes were compared between the postpolio patients and ten healthy controls.
When comparing levels in postpolio patients to the whole control group, including the 30 oldest investigated, there were no statistically significant differences. No difference was found in tumor necrosis factor levels induced by immune complexes when comparing patients and controls.
There was no increase in circulating immune complex or in tumor necrosis factor-inducing effects of circulating immune complex between postpolio patients and healthy controls, indicating that the postpolio syndrome is not due to an autoimmune reaction.
[Show abstract][Hide abstract] ABSTRACT: Early appearance of antibodies specific for native human type II collagen (anti-CII) characterizes an early inflammatory and destructive phenotype in adults with rheumatoid arthritis (RA). The objective of this study was to investigate the occurrence of anti-CII, IgM RF, IgA RF and anti-CCP in serum samples obtained early after diagnosis, and to relate the occurrence of autoantibodies to outcome after eight years of disease in children with juvenile idiopathic arthritis (JIA).
[Show abstract][Hide abstract] ABSTRACT: To investigate biomarker patterns in rheumatoid arthritis (RA) with extraarticular manifestations.
Cartilage oligomeric matrix protein (COMP), COMP-C3b, and soluble terminal complement complexes (sTCC) were measured by ELISA.
COMP-C3b levels were higher in patients with RA than in healthy controls and lower in extraarticular RA (ExRA) than in RA controls. In patients with ExRA, sTCC levels were higher than in RA controls, and correlated inversely with serum COMP-C3b levels in the ExRA group.
Patients with ExRA had lower levels of COMP-C3b. This may be a consequence of complement consumption or a lower potential for COMP from these patients to activate complement.
The Journal of Rheumatology 11/2013; · 3.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Introduction:
Analysis of antibodies against double-stranded DNA (anti-dsDNA) is important as a diagnostic and prognostic tool in systemic lupus erythematosus (SLE). Anti-dsDNA analysis by the Farr radio-immunoassay or the Crithidia luciliae immunofluorescence test (CLIFT) are regarded as reference methods. CLIFT has the advantage of offering IgG-specific antibody analysis, although the quantitative estimate by titration is rough compared to the Farr assay. Generally, anti-dsDNA analyses by conventional enzyme-immunoassays have lower diagnostic accuracy than CLIFT and Farr. Herein, CLIFT was used as reference to compare two more recent anti-dsDNA assays, i.e. the PhaDia enzyme immunoassay and the fluorescent microspehere immunodetection system (FIDIS).
Subjects and methods:
192 patients with SLE (93% Caucasians; 92% female; mean age 50.5 years/range 18-80; mean duration 11 years/range 0-45) participating in the KLURING study (Swedish acronym for ‘Clinical lupus register in southeast Gothia’). 81% of the patients met the 1982 ACR criteria, and the remainder fulfilled the Fries’ criteria, e.g. 2 typical organ manifestations at diagnosis and positive IgG antinuclear antibody (ANA/HEp-2) above the 95th percentile among 150 healthy female blood donors. For CLIFT and PhaDia, the following disease controls were included: 100 blood donors (50 women, 50 men), 97 with rheumatoid artritis and 54 patients with Sjögren’s syndrome. Sera were analyzed for the presence of IgG anti-dsDNA by CLIFT (cut-off >99th percentile among 100 healthy female blood donors) and by PhaDia (Thermo Fisher Scientific/Phadia AB) and FIDIS (Theradiag) with the cut-off limits suggested by the manufacturers.
When comparing anti-dsDNA results in relation to fulfillment of the ACR-82 criteria, Fisher’s exact test revealed that all three anti-dsDNA tests showed significant positive associations to criterion 7 (renal disease) and 10 (immunologic disorder). CLIFT and FIDIS, but not PhaDia, showed significant negative associations to criterion 3 (photosensitivity), whereas PhaDia alone showed a significant negative association to criterion 4 (oral ulcers). The proportions of positive anti-dsDNA tests in SLE, healthy subjects and disease controls are shown in Table 1.
Table 1. Positive anti-dsDNA tests obtained with CLIFT, Phadia and FIDIS.
SLE (n=192) RA (n=97) Sjögren (n=54) Healthy (n=100)
CLIFT 25 % 2 % 6 % 0 %
PhaDia 34 % 6 % 13 % 1 %
FIDIS 32 % nt nt nt
Spearman’s rho correlation revealed that anti-dsDNA levels measured by CLIFT correlated more strongly to FIDIS (rho 0.631, p<0.0005) than to PhaDia (rho 0.475, p<0.0005). The correlation between PhaDia and FIDIS was 0.607 (p<0.0005).
When CLIFT is chosen as ‘the gold standard’ for anti-dsDNA measurement in SLE, the present study indicates that FIDIS performs better than PhaDia. This applies both to the associations of positivie/negative tests in relation to disease manifestations, and to the serum levels of anti-dsDNA. As regards diagnostic specificities of the tests, we found that a positivie CLIFT was more SLE-specific than PhaDia. The corresponding results for FIDIS, however, remain to be elucidated.
The 10th international congress on SLE, Buenos Aires; 04/2013
[Show abstract][Hide abstract] ABSTRACT: We have previously reported that high levels of antibodies specific for native human type II collagen (anti-CII) at the time of RA diagnosis were associated with concurrent but not later signs of inflammation. This was associated with CII/anti-CII immune complex (IC)-induced production of pro-inflammatory cytokines in vitro. In contrast, anti-cyclic citrullinated peptide antibodies (anti-CCP) were associated both with late inflammation and late radiological destruction in the same RA cohort. We therefore hypothesized that anti-CII are also associated with early erosions.
Two-hundred-and-fifty-six patients from an early RA cohort were included. Baseline levels of anti-CII, anti-CCP and anti-mutated citrullinated vimentin were analyzed with ELISA, and rheumatoid factor levels were determined by nephelometry. Radiographs of hands and feet at baseline, after one and after two years were quantified using the 32-joints Larsen erosion score.
Levels of anti-CII were bimodally distributed in the RA cohort, with a small (3.1%, 8/256) group of very high outliers with a median level 87 times higher than the median for the healthy control group. Using a cut-off discriminating the outlier group that was associated with anti-CII IC-induced production of proinflammatory cytokines in vitro, baseline anti-CII antibodies were significantly (p = 0.0486) associated with increased radiographic damage at the time of diagnosis. Anti-CII-positive patient had also significantly increased HAQ score (p = 0.0303), CRP (p = 0.0026) and ESR (p = 0.0396) at the time of diagnosis but not during follow-up. The median age among anti-CII-positive subjects was 12 years higher than among the anti-CII-negative patients.
In contrary to anti-CCP, anti-CII-positive patients with RA have increased joint destruction and HAQ score at baseline. Anti-CII thus characterizes an early inflammatory/destructive phenotype, in contrast to the late appearance of an inflammatory/destructive phenotype in anti-CCP positive RA patients. The anti-CII phenotype might account for part of the elderly acute onset RA phenotype with rather good prognosis.
Arthritis research & therapy 05/2012; 14(3):R100. · 4.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The Study Group for Risk Factors for Rheumatoid Arthritis was established by the EULAR Standing Committee on Investigative Rheumatology to facilitate research into the preclinical and earliest clinically apparent phases of rheumatoid arthritis (RA). This report describes the recommendation for terminology to be used to define specific subgroups during different phases of disease, and defines the priorities for research in this area. Terminology was discussed by way of a three-stage structured process: A provisional list of descriptors for each of the possible phases preceding the diagnosis of RA were circulated to members of the study group for review and feedback. Anonymised comments from the members on this list were fed back to participants before a 2-day meeting. 18 participants met to discuss these data, agree terminologies and prioritise important research questions. The study group recommended that, in prospective studies, individuals without RA are described as having: genetic risk factors for RA; environmental risk factors for RA; systemic autoimmunity associated with RA; symptoms without clinical arthritis; unclassified arthritis; which may be used in a combinatorial manner. It was recommended that the prefix 'pre-RA with:' could be used before any/any combination of the five points above but only to describe retrospectively a phase that an individual had progressed through once it was known that they have developed RA. An approach to dating disease onset was recommended. In addition, important areas for research were proposed, including research of other tissues in which an adaptive immune response may be initiated, and the identification of additional risk factors and biomarkers for the development of RA, its progression and the development of extra-articular features. These recommendations provide guidance on approaches to describe phases before the development of RA that will facilitate communication between researchers and comparisons between studies. A number of research questions have been defined, requiring new cohorts to be established and new techniques to be developed to image and collect material from different sites.
Annals of the rheumatic diseases 03/2012; 71(5):638-41. · 8.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Production of anti-citrullinated protein antibodies (ACPAs) is an important biomarker for rheumatoid arthritis (RA). We undertook this study to determine whether genetic factors (HLA-DRB1 alleles) are associated with extreme ACPA levels in individuals with ACPA-positive RA, and to ascertain whether there are any phenotypic characteristics associated with these subgroups of RA.
HLA-DRB1 allelic groups were genotyped in 1,073 ACPA-positive RA patients from the Swedish Epidemiological Investigation of Rheumatoid Arthritis study. We found that 283 patients (26.4%) had high ACPA levels (defined as >1,500 units/ml using the Euro-Diagnostica anti-CCP2 test), while the rest of the patients had moderate ACPA levels and served as the comparison group. A replication group consisted of 235 RA patients.
No significant differences in baseline disease activity were observed between patients with high and those with moderate ACPA levels. However, the HLA-DRB1*15 allele was associated with high ACPA levels (P=0.0002). A similar trend was detected in HLA-DRB1*15-positive patients in the replication cohort, with meta-analysis of the discovery and replication cohorts demonstrating an overall effect of HLA-DRB1*15 on development of high ACPA levels in both the discovery and replication cohorts (P<0.0001 by Mantel-Haenszel test with a fixed-effects model).
Our data indicate that HLA-DRB1*15 may promote the production of high ACPA levels. Due to the high value of ACPA level scores in the 2010 American College of Rheumatology/European League Against Rheumatism classification criteria for RA, the presence of HLA-DRB1*15 may, at least in part, contribute to fulfilling the criteria for RA. This illustrates the complex nature of the genetic regulation of ACPA levels. Additional mechanistic studies of the regulation of ACPAs and ACPA-positive RA are pending.