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Systemic lupus erythematosus (SLE) most commonly manifests as mild to moderate disease with severe manifestations such as diffuse alveolar hemorrhage, central nervous system vasculitis, macrophage activation syndrome (MAS) or retinal vasculitis (RV) with visual disturbances occurring in a significantly smaller proportion of patients, most of whom have a poor outcome. Macrophage activation syndrome and RV are insufficiently early and rarely recognized presentations of lupus—consequently there are still no treatment recommendations. Here we present the course of diagnosis and treatment of a patient with an SLE flare that resulted in both life-threatening disease (MAS) and vision-threatening disease (RV). The patient was successfully treated with systemic immunosuppressives, a high dose of glucocorticoids and rituximab (RTX), in parallel with intraocular therapy, intravitreal bevacizumab (BEV) and laser photocoagulation.

Aim: To investigate the association of FasL gene polymorphism (rs763110) with rheumatoid arthritis occurrence, disease activity, and tumor necrosis factor-α (TNF-α) plasma concentration in Croatian patients, and to conduct an updated meta-analysis. ----- Methods: This cross-sectional study enrolled 81 patients with rheumatoid arthritis and 94 control patients. After the assessment of the Disease Activity Score (DAS)-28, blood was taken for analysis. DNA was isolated from the whole blood to determine FasL polymorphism (rs763110) by polymerase chain reaction. Protein levels of TNF-α were determined with ELISA. After a detailed literature search, we conducted an updated meta-analysis using the Review Manager 5 software. ----- Results: Rheumatoid arthritis patients had significantly higher TNF-α concentration in plasma (1.65 [1.2-2.42] pg/mL) than controls (0.99 [0.77-1.35] pg/mL, P

Rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA) are associated with abnormal immune cell functions. We combined manual and automated profiling in subpopulations of T-cells, B-cells and monocytes, in parallel to functional testing and clinical correlation.Using flow cytometry, we analysed the expression of CCR4, CCR6 and CXCR5 on helper and cyotoxic T-cells, CD32B and CD86 on naïve and memory B-cells, and CCR1, CCR2, CCR4 and CXCR4 on monocytes in chronic high-disease activity patients to identify peripheral blood subpopulations. Cell activation, proliferative capability and osteoclastogenic effects were tested in vitro. Comparison with synovial compartment, clinical data and anti-TNF treatment were added to peripheral blood analysis.PsA had lower double-negative T-cell frequency, while RA had lower double-positive T-cell frequency and expanded Th1-like and cytotoxic T-cell subsets. CD32B expression was increased on naïve and memory B-cells in AS and associated with disease activity. CCR6+ and CXCR5+ cytotoxic T-cells and CD32B+ naïve and memory B-cells were highly enriched within the synovial compartment. T-cells and B-cells from AS exhibited enhanced activation and proliferation in vitro, whereas T-cell conditioned medium from RA produced an increased osteoclastogenic effect. CCR1 and CXCR4 were upregulated on osteoclastogenic monocyte subsets of RA, AS and PsA patients. Bioinformatic Citrus analysis identified additional T-cell, B-cell and monocyte clusters specifically associated with each disease.By combining manual and automated data analysis, our study revealed several disease-specific immune cell subpopulations, particularly cytotoxic T-cell subsets in RA and memory B-cell subsets in AS, which may serve as an indicator of active disease or possible therapeutic target.

Background: Autoimmunity is major driving force in pathogenesis of rheumatic diseases, including ankylosing spondylitis (AS). AS is associated with abnormal immune cell functions. Several recent studies stressed the role of T- cells and monocyte subpopulations in AS, but these mechanisms are signifi cantly less understood compared to other rheumatic diseases (1). Objectives: Th e aim was to compare the frequency of T-cell and monocyte subpopulations in AS patients and controls, and to correlate them with disease activity. Methods: Mononuclear cells (MNC) were isolated from blood of healthy controls (n=110) and AS (n=65) patients. T-cell and monocyte phenotype of MNC was determined using fl ow cytometry for the following markers: Th 1/2 (CD3+CD4+CCR6–), Th 17 (CD3+CD4+CCR4+CCR6+), Tfh (CD3+CD4+CXCR5+), Tc (CD3+CD8+), memory Tc (CD3+CD8+CCR4+) double- positive (dp) Tc (CD3+CD4+CD8+) ; and monocytes (CD3-CD19-CD56-CD11b+CD14+). On lymphocytes IL21R, CD25 and CD11b were analyzed ; while on monocytes chemokine receptors CCR1, CCR2, CCR4, CXCR4 and RANK. Frequencies of lymphocyte and monocyte subpopulations were correlated with ASDAS, BASDAI, VAS of pain in spine and peripheral joints, stiff ness intensity and duration, ESR and CRP. Results: Results showed decrease of memory Tc (p=0.022) and Tfh in AS (p=0.038) which correlated negatively with physician’s assessment of disease activity. Th e frequency of dp Tc was unchanged, but negatively correlated with BASDAI and pain assessment. Th e frequency of monocytes was decreased in AS (p

Background The peripheral blood (PB) monocyte pool contains osteoclast progenitors (OCPs), which contribute to osteoresorption in inflammatory arthritides and are influenced by the cytokine and chemokine milieu. We aimed to define the importance of chemokine signals for migration and activation of OCPs in rheumatoid arthritis (RA) and psoriatic arthritis (PsA). Methods PB and, when applicable, synovial fluid (SF) samples were collected from 129 patients with RA, 53 patients with PsA, and 110 control patients in parallel to clinical parameters of disease activity, autoantibody levels, and applied therapy. Receptors for osteoclastogenic factors (CD115 and receptor activator of nuclear factor-κB [RANK]) and selected chemokines (CC chemokine receptor 1 [CCR1], CCR2, CCR4, CXC chemokine receptor 3 [CXCR3], CXCR4) were determined in an OCP-rich subpopulation (CD3−CD19−CD56−CD11b+CD14+) by flow cytometry. In parallel, levels of CC chemokine ligand 2 (CCL2), CCL3, CCL4, CCL5, CXC chemokine ligand 9 (CXCL9), CXCL10, and CXCL12 were measured using cytometric bead array or enzyme-linked immunosorbent assay. Sorted OCPs were stimulated in culture by macrophage colony-stimulating factor and receptor activator of nuclear factor-κB ligand, and they were differentiated into mature osteoclasts that resorb bone. Selected chemokines (CCL2, CCL5, CXCL10, and CXCL12) were tested for their osteoclastogenic and chemotactic effects on circulatory OCPs in vitro. Results The OCP population was moderately enlarged among PB cells in RA and correlated with levels of tumor necrosis factor-α (TNF-α), rheumatoid factor, CCL2, and CCL5. Compared with PB, the RANK+ subpopulation was expanded in SF and correlated with the number of tender joints. Patients with PsA could be distinguished by increased RANK expression rather than total OCP population. OCPs from patients with arthritis had higher expression of CCR1, CCR2, CCR4, CXCR3, and CXCR4. In parallel, patients with RA had increased levels of CCL2, CCL3, CCL4, CCL5, CXCL9, and CXCL10, with significant elevation in SF vs PB for CXCL10. The subset expressing CXCR4 positively correlated with TNF-α, bone resorption marker, and rheumatoid factor, and it was reduced in patients treated with disease-modifying antirheumatic drugs. The CCR4+ subset showed a significant negative trend during anti-TNF treatment. CCL2, CCL5, and CXCL10 had similar osteoclastogenic effects, with CCL5 showing the greatest chemotactic action on OCPs. Conclusions In our study, we identified distinct effects of selected chemokines on stimulation of OCP mobilization, tissue homing, and maturation. Novel insights into migratory behaviors and functional properties of circulatory OCPs in response to chemotactic signals could open ways to new therapeutic targets in RA. Electronic supplementary material The online version of this article (doi:10.1186/s13075-017-1337-6) contains supplementary material, which is available to authorized users.

Background Both rheumatoid arthritis (RA) and ankylosing spondylitis (AS) belong to the group of chronic rheumatic diseases with the important role of autoimmune pathogenic mechanisms. However, these forms differ in the major target tissues as well as the intensity of bone and cartilage destruction, with RA being the prototype of “destructive” arthritis affecting peripheral joints and AS being the prototype of “remodeling” arthritis predominantly of the axial skeleton. Objectives Both forms of arthritis are associated with abnormal immune cell functions, including B lymphocytes. Although several recent studies stressed the role of B lymphocyte subpopulations and autoantibodies in AS, these mechanisms in the AS pathogenesis are significantly less understood compared to RA and other rheumatic autoimmune diseases.The aim of our study was to compare the frequency of B lymphocyte subpopulations between RA and AS patients, and to correlate them with the disease activity. Methods Mononuclear cells were isolated from peripheral blood of healthy controls (n=30), RA (n=33) and AS (n=18) patients, after obtaining Ethical approval and informed consent from patients. The B lymphocyte phenotype of peripheral-blood mononuclear cells was determined using flow cytometry for the following surface markers: CD19, CD27, IgD, CD32 and CD38. In addition, statistical correlation was assessed to test the association between the frequency of selected B lymphocyte subpopulations and the disease-activity score, DAS28 (Disease Activity Score 28) for RA and BASDAI (Bath Ankylosing Spondylitis Disease Activity Index) for AS. Results Gating strategy applied for flow- cytometry data was set to discriminate between naïve B lymphocytes (CD19+ IgD+ CD27-), unswitched memory B lymphocytes (CD19+ IgD+ CD27+), class-switched memory B lymphocytes (CD19+ IgD- CD27-) and plasmablasts (CD19+ IgD+ CD27hi CD38+). In addition, expression of CD32 (FcgRII receptor) and CD86 (B7-2 co- stimulator), associated with the maturation and activation of B lymphocytes, within these B lymphocyte subsets were assessed. Data analysis showed expanded CD32+ subset among naïve and memory B lymphocytes in RA (16.4±11.6% and 9.8±8.2%) compared to control (4.3±2.7% and 7.6±3.5%) and AS (5.6±2.6% and 5.8±1.9%). Similarly, there were more CD86+ cells among naïve and unswitched memory B lymphocytes in RA (9.0±8.2% and 18.2±8.0%) compared to control (2.8±1.3% and 11.7±5.5%) and AS (4.9±5.9% and 11.6±6.2%). Class-switched memory B lymphocytes were negatively associated with disease activity score in RA (ρ=-0.45) but positively in AS (ρ=+0.39), whereas the associations were reversed for the population of naïve B lymphocytes (ρ=+0.32 for RA and ρ=-0.38 for AS). Conclusions Our results indicate that B lymphocyte-mediated immune response may be important for both RA and AS, but with distinct effector mechanisms. Therefore it is reasonable to suggest that B lymphocyte subpopulations may represent promising cellular targets for the therapeutic interventions in different forms of arthritis.

Rheumatoid arthritis (RA) is marked by persistent inflammation and joint destruction involving infiltration of osteoclasts. Human osteoclast progenitors (OCPs) are contained among peripheral blood monocytes at low frequency, exhibit chemotaxis and, furthermore, synovial compartment of RA patients highly expresses different chemokines. The aim was to define these chemotactic signals. Mononuclear cells were isolated from peripheral blood of control and RA patients. The phenotype of OCPs (CD3-CD19-CD56-CD11b CD14 ) was determined using flow cytometry for receptors CCR1, CCR2, CCR4, CXCR4, C5AR1. Chemokine ligand concentrations (CCL2, CCL3, CCL4, CCL5, CXCL9, CXCL10) were measured in serum and synovial fluid of RA patients using cytometric bead assay. OCPs were sorted and cultured with M-CSF and RANKL. After two weeks, the cells were stained for TRAP enzyme and mature osteoclasts were counted. Human peripheral blood OCPs similarly expressed chemokine receptors in RA and healthy subjects. However, CCL4 and CXCL10 concentrations were significantly higher in synovial fluid compared to serum levels in RA, while CCL2, CXCL9 and CXCL10 serum levels were higher in RA patients compared to the control group. Cell culture revealed no significant differences in mature osteoclast count between groups. Although OCPs in RA have a differentiation potential similar to controls, levels of several chemokines are upregulated, indicating a possible chemotactic mechanism of OCP migration to affected joints. These results may help to reveal a migration mechanism of OCPs specifically associated with RA in order to develop more efficient therapeutic approaches.

U posljednjih nekoliko godina u uporabu su uvedeni mnogi novi lijekovi, stvarajući nove obrasce preosjetljivosti. Uvode se novi klinički i biološki postupci koji omogućavaju razumijevanje mehanizama i dijagnosticiranje reakcija na stare generacije lijekova. Ovo je pregled znanja o epidemiologiji, mehanizmima te kliničkoj i in-vitro dijagnostici tih reakcija. Nedavna otkrića ukazuju da tradiocionalni i alternativni lijekovi također mogu uzrokovati neželjene reakcije, od kojih se mnoge obilježavaju kao alergijske. Istraživanja u području kožnih i provokacijskih testova na antibiotike, kao što su betalaktami i karbapenemi, te jodirane i gadolinij kontrastne medije omogućila su razumijevanje križne reaktivnosti i sigurnosti primjene zamjenskih lijekova s dobrom podnošljivošću u slučaju adekvatno provedene alergološke obrade. Zabilježeni su mnogi jedinstveni slučajevi koji uključuju različite lijekove poput infliximaba, sukcinilkolina, hidroksiklorokina, koji proširuju spektar kliničkih manifestacija hipersenzitivnih reakcija na lijekove. Nekoliko objavljenih studija iz područja in-vitro dijagnostike, uključujući test aktivacije bazofila (BAT-engl. basofil activation test), radioalergosorbentni test, imuno-Cap, ELISA-u, enzim-linked immunospot test i testove proliferacije T-stanica predstavljaju nove alate za procjenu alergije na lijekove. Upotrebom novih i starih lijekova dolazimo do izvještaja o novim i ranije poznatim manifestacijama hipersenzitivnosti na lijekove. Napredak u razumijevanju mehanizama omogućen je uporabom novih in-vitro testova za detekciju specifičnih protutijela ili T-limfocita. Istraživanja u području kožnih i provokacijskih dijagnostičkih postupaka olakšavaju odluke o sigurnosti upotrebe zamjenskih lijekova kod osoba s dokazanom medikamentoznom alergijom