Lack of measures of disease activity and prognosis is a key challenge in spondyloarthritis (SpA). Several soluble biomarkers have been proposed, but very few have been found to relate to MRI activity and add to the diagnostic value of CRP.(1) Leukocytes are key players in SpA disease activity.(2) The soluble form of CD18 (sCD18) has been found to be shed from leukocytes in inflammation and to be present in the blood.(3)
Objectives
This study examines the association of sCD18 in the blood and SpA disease activity.
Methods
Plasma from a study population with 84 SpA patients at baseline and 33 patients at three-year follow-up and a control group of age and gender matched normal healthy volunteers (NHV) were studied. The samples were analyzed for sCD18 by a time resolved immunoflourometric assay (TRIFMA) using the KIM185 antibody as capture antibody and the KIM127 antibody as detection antibody as previously described.(3)
Results
The level of sCD18 was decreased in plasma from SpA patients compared to plasma from NHV (p<0.0005). The level of sCD18 showed inverse correlations to the clinical parameters clinician defined disease activity on a 100-mm visual analog scale (VAS) (p<0.05) and Bath Ankylosing Spondylitis Metrology Index (BASMI) (p<0.005) and the paraclinical measures CRP (p<0.005) and sacroiliac joint (SIJ) MRI activity score (p<0.05). Interestingly, CRP did not show any significant relation to clinical findings or MRI activity in this study. Patients with BASMI improvement at three-year follow-up also had increases in sCD18 concentration compared to patients without BASMI improvements (p<0.05).
Conclusions
sCD18 is associated with clinical findings, CRP and SIJ MRI activity in SpA. sCD18 could be a novel disease activity marker in SpA directly measuring leukocyte activity.
References
Maksymowych WP. Biomarkers in spondyloarthritis. Current rheumatology reports. 2010;12(5):318-24. Dougados M, Baeten D. Spondyloarthritis. Lancet. 2011;377(9783):2127-37. Gjelstrup LC, Boesen T, Kragstrup TW, Jorgensen A, Klein NJ, Thiel S, et al. Shedding of large functionally active CD11/CD18 Integrin complexes from leukocyte membranes during synovial inflammation distinguishes three types of arthritis through differential epitope exposure. J Immunol. 2010;185(7):4154-68.
Systemic sclerosis (SSc) is an autoimmune disease cha-racterized by vascular alterations, immune activation, andfibrosis in multipleorgans. T cellsdominateinskinlesions,withCD4Tcellsasthemajor subpopulation.TheseCD4Tcells display markers of activation such as the solubleinterleukin 2 receptor (sIL-2R), and exhibit oligoclonalexpansion (1–3). The CTLA-4 domain on abatacept bindstoCD80/CD86onantigen-presentingcellsinhibitingCD28from binding on T cells and consequently inhibiting T-cellactivation (4). This led us to use abatacept in patientssuffering from treatment-resistant diffuse cutaneous SSc(dcSSc).Abatacept was added to conventional therapy in fourpatients suffering from severe and treatment-resistantdcSSc. All patients met classification criteria for SSc(5). Abatacept was administered intravenously (weight< 60 kg: 500 mg, 60–100 kg: 750 mg) at weeks 0, 2,and 4, and then every 4 weeks.Patient A, a 21-year-old anti-topoisomerase-positivewoman with Raynaud’s phenomenon (February 2007),reducedoralaperture,oedemaofthehandsandarms,severethickening of the skin on extremities and trunk, and con-tractures of elbows and knees was admitted in November2007. Despite initial therapy with methotrexate and pred-nisolone followed by oral cyclophosphamide and pred-nisolone, the modified Rodnan skin score (mRSS)increased, carbon monoxide diffusing capacity (DLCO)decreased to 68% of predicted (Figures 1A and 1C), andthe patient experienced shortness of breath [New YorkHeart Association (NYHA) classification III], tiredness,and muscle weakness just able to walk 15 min. beforeresting. Echocardiography and high-resolution computedtomography (HRCT) of the lungs were normal. Abatacepttreatment was added in May 2008. During the followingmonths, subjective symptoms, mobility of the fingers,mRSS,andprocollagen-3N-terminalpeptide(P3NP)levelsall improved (Figure 1A). All treatments were terminatedduring August and September 2009. By November 2010,lung function had improved and the woman performed 532m in the six-minute walk test (6MWT) (Figure 1C). P3NPlevelsandmRSScontinuedtofall.Patient B, a 55-year-old, anti-topoisomerase-positivewoman, who developed dcSSc in January 2009, wasadmitted in October 2009 with swelling of fingers, stiffnessofskin,interstitiallungdisease(ILD),pericarditis,suspectedmyocarditis, oesophagus hypomotility, weight loss, andpolymyositis. Within a few weeks the patient developedscleroderma renal crisis and a complicated pneumonia,causing cyclophosphamide treatment to cease after 2weeks. Kidney function was stabilized with angiotensin-converting enzyme (ACE) inh ibitor. Skin thickening andILD progressed (NYHA III) (Figures 1B, 1D, and 1E). Herpoorgeneralconditionandtheriskofanewsevereinfectionprohibited further treatment with cyclophosphamide. OnDecember 2009 abatacept treatment was added to low-doseprednisolone. Creatinine (CK) and troponin-T (TNT) levelspromptly normalized and the symptoms of polymyositisreduced. In June 2010 mycophenolate mofetil was added.After months of progression, skin symptoms improvedalongwithanabruptdecreaseinP3NP,TNT,andCKandregressionofILDchangesonHRCT.Hergeneralconditionimproved; from being bedridden for 5 months she was ableto perform 250 m in the 6MWT.Patient C, a 48-year-old, anti-RNA polymerase I,IIIantibody-positive woman with oedema of the hands(January 2009) was diagnosed with dcSSc in May 2009with severe thickening of the skin on the extremities andabdomen, hyperalgesia, and dyspnoea during moderateexercise. Despite therapy with intravenous and later oralcyclophosphamide and prednisolone, the progressioncontinued. In August 2009 mRSS was 35. UVA-1 treat-ment was added for 3 months. Despite no change inmRSS, new areas not previously affected evolved.P3NP and urine protein excretion were increased to 8.9μg/L (1.7–4.2 μg/L) and 208 mg/day, respectively. InMay 2010, abatacept was added. Hereafter mRSS andP3NP fell, a persistently elevated C-reactive protein(CRP) level normalized, and proteinuria disappeared(Figure 2C). Cyclophosphamide was reduced and inAugust 2010 replaced by azathioprine. The patient’swell-being and functionality improved along with a con-tinuous decrease in mRSS and P3NP.
The aminoterminal propeptide of type III procollagen (PIIINP) and the carboxyterminal propeptide of type I procollagen (PICP) and hyaluronan (HA) were measured in plasma and suction blister fluid from 13 systemic sclerosis patients and 11 healthy volunteers. Suction blisters and skin biopsies were from the transition zone between normal skin and scleroderma, and uninvolved abdominal skin of patients. The median value of suction blister PIIINP from the transition zone was 38% higher than suction blister PIIINP from uninvolved skin. PIIINP was localized to the dermis by immunohistochemical techniques. PICP and HA levels in blisters from the transition zone were 87% and 53%, respectively, above the levels measured in uninvolved skin. Furthermore, PICP and HA blister levels from the transition zone were 67% and 63%, respectively, higher than the levels measured in healthy volunteers. In plasma from scleroderma patients levels of PIIINP and HA were 38% and 127% higher, respectively, than in plasma of healthy volunteers. The plasma PICP level was not significantly higher in scleroderma patients. Finally, PICP, PIIINP and HA levels were several times higher in suction blister fluid than in plasma. The data indicate that a fibrogenetic process takes place in the transition zone of scleroderma. The method may be used to monitor the progression of scleroderma skin lesions in vivo.
Rheumatoid arthritis (RA) is the most prevalent autoimmune mediated joint disease. A complex dysfunction of the immune response is considered central for its pathogenesis as well as a dysregulated crosstalk between fibroblast-like synoviocytes (FLS) and monocytes.
Objectives
This project aims to examine and validate the anti-inflammatory effects of the natural glucose metabolite itaconate-derivative 4-octyl itaconate (4-OI) on stromal and immune cells from patients with active RA and evaluate the downstream effector molecule Heme-Oxygenase 1 (HO-1) as a potential biomarker.
Methods
Synovial fluid mononuclear cells (SFMC) from patients with chronic RA (cRA) were used to harvest RA FLS. Monocultures of synovial fluid derived FLS (SF-FLS) and autologous co-cultures of SF-FLS and peripheral blood mononuclear cells (PBMC) were cultured with and without 4-OI, corticosteroids, and anti-TNF (n=7). Subsequently, analyses by flow cytometry, Western Blotting, MTT assay, and ELISA were performed. The 4-OI target protein Nrf2 was knocked out in immortalized FLS (FLS-KO) generated by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). Furthermore, we measured HO-1 by ELISA in plasma samples from newly diagnosed, active and treatment-naïve early RA (eRA) patients at baseline and after 6 months of intensive treatment (the OPERA trial, n=80)[1] with a follow-up period of 24 months and plasma and in synovial fluid samples from patients with active, chronic RA (cRA, n=20) during disease activity flares. Plasma samples from age approximated and gender matched healthy controls (HC) (n=35) were also included.
Results
In vitro 4-OI significantly inhibited MCP-1 secretion in cultured SFMC, THP-1 (monocyte cell line), and FLS and resulted in a concomitant increased HO-1 production. 4-OI reduced the MCP-1 secretion by 84%, anti-TNF by 41% and corticosteroids by 54% (all p<0.01) in co-cultures consisting of autologous RA SF-FLS and PBMCs. In FLS we observed a linear decrease in pro-inflammatory cytokine production. 4-OI did not decrease collagen production, even in dosage of 500 µn FLS we observed a linear decrease in pro-inflammatory cytokine production. 4-OI did not decrease collagen production, even in dosage of 500 1 production. 4-OI reduced the MCP. In eRA plasma levels of HO-1 was significantly increased at baseline (2039 pg/mL, 95% CI [1619, 2460]) vs HC (1209 pg/mL, 95% CI [1047, 1370]). Treatment with MTX or MTX + anti-TNF, lowered levels of HO-1 (1704 pg/mL, 95% CI [1377, 2031]) after 6 months irrespective addition of anti-TNF treatment, reducing the difference to a no longer significantly different level from HCs. HO-1 was associated with number of swollen joints (r=0.25, p<0.02), tender joints (r=0.24, p<0.02) and Clinical Disease Activity Index (r=0.23, p<0.02). Baseline HO-1 predicted radiographic joint damage progression (change in Total Sharp-van der Heijde score (Δn eRA plasma levels of HO-1 was significantly increased at baseline (2039 pg/mL, 95% CI [1619, 2460]) vs HC (1209 pg/mL, 95% CI [1047, 1370]). Treatment with MTX or MTX + anti-TNF, lowered levels of HO-1 (1704 pg/mL, 95% CI [1377, 2031]) after 6 months irund (r=0.75, p<0.001). High levels of HO-1 in SF from inflamed joints were associated with CRP (r=0.52, p<0.05) during flares in cRA.
Conclusion
In vitro 4-OI decreased MCP-1 secretion and increased the HO-1 expression in cultures of monocytes and in RA FLS and PBMC without decrease in collagen production or cell growth. In total, 4-OI decreased MCP-1 in both immune and stromal dominated RA pathobiology. In eRA patients, pre-treatment-plasma HO-1 levels were increased and correlated with parameters for disease activity, treatment-effect after 6 months, and predicted radiographic progression after 24-month.
Reference
[1] Hørslev-Petersen K et al. Ann Rheum Dis. 2013
Acknowledgements
We thank medical doctors and nurses at the Department of Rheumatology, Aarhus University Hospital for helping to collect the patient samples. The authors kindly acknowledge a generous grant from the Danish Rheumatoid Association (R177-A6156).
In osteoimmunology, osteoclastogenesis is understood in the context of the immune system. Today, the in vitro model for osteoclastogenesis necessitates the addition of recombinant human receptor activator of nuclear factor kappa‐B ligand ( RANKL ) and macrophage colony‐stimulating factor (M‐ CSF ). The peripheral joints of patients with rheumatoid arthritis ( RA ) and spondyloarthritis (SpA) are characterized by an immune‐mediated inflammation that can lead to bone destruction. Here, we evaluate spontaneous in vitro osteoclastogenesis in cultures of synovial fluid mononuclear cells ( SFMC s) activated only in vivo . SFMC s were isolated and cultured for 21 days at 0.5–1.0 × 10 6 cells/mL in culture medium. SFMC s and healthy control peripheral blood monocytes were cultured with RANKL and M‐ CSF as controls. Tartrate‐resistant acid phosphatase ( TRAP ) positive multinucleated cells were found in the SFMC cultures after 21 days. These cells expressed the osteoclast genes calcitonin receptor, cathepsin K, and integrin β3, formed lacunae on dentin plates and secreted matrix metalloproteinase 9 ( MMP 9) and TRAP . Adding RANKL and M‐ CSF potentiated this secretion. In conclusion, we show that SFMC s from inflamed peripheral joints can spontaneously develop into functionally active osteoclasts ex vivo . Our study provides a simple in vitro model for studying inflammatory osteoclastogenesis.
To examine the overall cognitive development of children exposed to maternal rheumatoid arthritis (RA) in utero by comparing their school test scores to those of their peers.Children born in Denmark during 1995-2008 and listed in the National School Test Register were included (n = 738,862). Children exposed to maternal RA were identified through linkage of national registers. In separate analyses, exposure was subdivided according to maternal serostatus. Preclinical maternal RA was included as a separate exposure. The Danish national school tests are mandatory standardized tests. Results from all reading tests (grades 2, 4, 6, and 8) and mathematics tests (grades 3 and 6) from 2010-2017 were included. Test scores were compared according to maternal RA exposure for each test separately using linear regressions.We identified 934 children exposed to maternal RA in utero. There were no differences in reading test scores between maternal RA exposed and unexposed children. RA exposed children scored poorer in both mathematics tests (adjusted differences of mean score -0.14 SD (95% confidence interval [95% CI] -0.23, -0.06) and -0.16 SD (95% CI -0.26, -0.07). There was no appreciable difference between children by maternal RA serostatus. Children exposed to preclinical RA (n = 589) showed the same pattern of performance as children exposed to RA.RA-exposed children scored slightly poorer in mathematics tests but performed as well as their unexposed peers in the reading tests. The results do not suggest that RA in pregnancy has a major impact on offspring school performance.
Abstract: Caspase 14 is a unique member of the cysteinyl aspartate-specific proteinase family. Its expression is confined primarily to cornified epithelium such as the skin. Caspase 14 has been associated with the processing of filaggrin monomers and the development of natural moisturising factors of the skin, and thus, it could be speculated that caspase 14 dysregulation is implicated in the development of an impaired skin barrier function. We have investigated the regulation of caspase 14 transcription in cultured primary keratinocytes following stimulation with a number of factors present in inflamed skin, including TH1- and TH2-associated cytokines in addition to LPS and peptidoglycan. In particular, we found that TH2-associated cytokines reduced the caspase 14 mRNA level significantly. Furthermore, we found that the expression of caspase 14 was reduced in skin biopsies from patients with atopic dermatitis (AD), psoriasis and contact dermatitis, further supporting a role for this kinase in inflammatory skin conditions. Hence, the regulation of caspase 14 levels provides a possible link between impaired skin barrier function and inflammatory reactions in skin diseases such as AD and may offer an explanation to the skin barrier dysfunction in inflamed skin lesions.
Recent studies suggest that familial autoimmunity plays a part in the pathogenesis of ASDs. In this study we investigated the association between family history of autoimmune diseases (ADs) and ASDs/infantile autism. We perform confirmatory analyses based on results from previous studies, as well as various explorative analyses.The study cohort consisted of all of the children born in Denmark from 1993 through 2004 (689 196 children). Outcome data consisted of both inpatient and outpatient diagnoses reported to the Danish National Psychiatric Registry. Information on ADs in parents and siblings of the cohort members was obtained from the Danish National Hospital Register. The incidence rate ratio of autism was estimated by using log-linear Poisson regression.A total of 3325 children were diagnosed with ASDs, of which 1089 had an infantile autism diagnosis. Increased risk of ASDs was observed for children with a maternal history of rheumatoid arthritis and celiac disease. Also, increased risk of infantile autism was observed for children with a family history of type 1 diabetes.Associations regarding family history of type 1 diabetes and infantile autism and maternal history of rheumatoid arthritis and ASDs were confirmed from previous studies. A significant association between maternal history of celiac disease and ASDs was observed for the first time. The observed associations between familial autoimmunity and ASDs/infantile autism are probably attributable to a combination of a common genetic background and a possible prenatal antibody exposure or alteration in fetal environment during pregnancy.
Allergy and allergic diseases have increased in prevalence worldwide during the last decade. Relevant determinants influencing the development of allergic inflammation come from the environment and are either enhancing (e.g. environmental pollutants both indoors and outdoors) or protective (e.g. parasite infestations causing early stimulation of the immune system). In spite of considerable progress in experimental allergology and immunology, there is still a great discrepancy between theoretical knowledge and practical performance in the routine treatment of patients with allergies. The development of new therapeutic and preventive strategies for the future management of allergy is dependent on a better understanding of the pathomechanisms and molecular pathways involved. Based on an international symposium, this volume summarizes the latest findings in epidemiology, pathophysiology, and clinical aspects of allergic diseases such as asthma, food allergy, and, especially, atopic eczema. Risk factors for the development of allergies and novel treatment strategies are carefully evaluated. This update is essential reading for anyone interested in allergy: doctors working in the clinical fields of dermatology, pneumology, internal medicine, pediatrics, ENT, epidemiology and public health, as well as researchers in molecular genetics, immunobiology, food and nutrition sciences, and pharmacology.
