BackgroundOur aim was to compare serum levels of cytokines in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA).ObjectivesComparative data on cytokine profile in SLE and rheumatoid arthritis RA patients are scarce.MethodsWe examined serum samples from 80 pts with SLE, median and interquartile range (25th–75th percentile) of disease duration 48 (2–432) months; age 31,5 (16–65) years; 72 female; 74 pts with RA, disease duration 90,0 (30,00–192,0) months; age 54,0 (44,0–62,0) years; 59 female, and 28 healthy donors. Cytokine analyses were performed with Bio-Plex® technology (Human Grp I Cytokine 27-plex panel).ResultsPts with SLE had significantly lower levels of IL-1β, -1ra, -2, -9, eotaxin, G-CSF, IFN-γ, MIP-1β, TNF-α, VEGF and higher levels of IL-4, -6, -8, -12, GM-CSF, MCP-1, PDGF-BB, RANTES than healthy donors. Compared to RA, cytokine/chemokine levels from SLE were significantly different. The concentrations of IL-1β, -1ra, -2, -5, -6, -7, -9, -10, -13, 15, eotaxin, FGF, G-CSF, IFN-γ, IP-10, MIP-1α, TNF-α, VEGF in SLE were significantly lower than RA. The concentrations of IL-4, -8, MCP-1, MIP-1β, PDGF-BB, RANTES was higher in the SLE cohort (Table 1).ConclusionsSerum concentration of most proinflammatory, Th-2 related, bone marrow–derived cytokines, stromal cells and angiogenic factors in SLE pts is substantially lower than in healthy donors and pts witch RA. These data demonstrate significantly higher chemokine levels in SLE versus RA.ReferencesChun HY, Chung JW, Kim HA, Yun JM, Jeon JY, Ye YM, Kim SH, Park HS, Suh CH. Cytokine IL-6 and IL-10 as biomarkers in systemic lupus erythematosus. J Clin Immunol. 2007; 27(5):461–6.Kokkonen H, Söderström I, Rocklöv J, Hallmans G, Lejon K, Rantapää Dahlqvist S. Up-regulation of cytokines and chemokines predates the onset of rheumatoid arthritis. Arthritis Rheum. 2010; 62(2):383–91. doi: 10.1002/art.27186.Sieber J, Daridon C, Fleischer SJ, Fleischer V, Hiepe F, Alexander T, Heine G, Burmester GR, Fillatreau S, Dörner T. Active systemic lupus erythematosus is associated with a reduced cytokine production by B cells in response to TLR9 stimulation. Arthritis Res Ther. 2014;16(6):477. doi: 10.1186/s13075-014-0477-1.Disclosure of InterestNone declared
BackgroundAutoantibodies against intracellular antigens are a serological hallmark of ANA-associated systemic autoimmune rheumatic diseases (AARD) such as SLE. IIF on HEp-2 cells for ANA remains the “gold standard” but it has very low positive predictive value. Up to 20% of serum samples from HI have been reported to have a positive ANA IIF test, the majority of them due to the presence of anti-dense fine speckled 70 (anti-DFS70) antibodies. Monospecific anti-DFS70 antibodies represent a biomarker that can be used to discriminate AARD patients (pts) from HI in ANA IIF positive subjects. Recognition of the DFS70 ANA IIF pattern can be challenging. The DFS-KO Hep-2 cells inhibit anti-DFS70 antibodies reactions, providing clear differentiation of the DFS pattern from classical ANA patterns.ObjectivesTo evaluate the utility of a novel HEp-2/DFS70-KO IIF substrate for the detection of anti-DFS70 antibodies in HI and SLE pts.MethodsWe studied 45 HI (36 F/9 M; age 50.4 [24.0–72.0] years, median [interquartile range 25–75%]) and 12 pts with SLE (ACR criteria, 1997) (10 F/2M, age 38.9 [17.0–65.0] years; disease duration 100.3 [4.0–432.0] months; SLEDAI 2K score 11.7 [2–30]; SLICC damage index score 1.28 [0–4]). Serum samples were tested for classical ANA and anti-DFS70 antibodies by IIF technique with a mixture of standard HEp-2 cells and DFS70-KO HEp-2 cells (“Trinity Biotech”, Bray, Ireland) as a substrate. Fluorescence titers ≥1: 160 were considered as positive for ANA patterns.ResultsANA were present in 7/45 (15.6%) of HI and in 12/12 (100%) of SLE pts. All SLE pts and 3/45 (6.7%) of HI showed classic ANA patterns (homogeneous, speckled, and mixed) in the absence of DFS70 pattern. 4/45 (8.9%) of HI had classic ANA negative/anti-DFS70 antibodies positive IIF results. Isolated anti-DFS70 antibodies were found in 57% of ANA IIF positive HI. Among HI classic ANA and anti-DFS70 antibodies were detected in the low- to medium-titer range (1:160–1:320). The frequency of anti-DFS70 antibodies did not correlated with age.ConclusionsThe detection of isolated anti-DFS70 antibodies may be regarded as an exclusion criterion for the diagnosis of SLE. The testing for anti-DFS70 antibodies in a single step by HEp-2/DFS70-KO IIF method should be included into a modified ANA diagnostic algorithm. Additional investigations are required to evaluate the clinical relevance of anti-DFS70 autoantibodies.Disclosure of InterestNone declared
Background The B lymphocyte stimulator signaling pathway by BAFF and its homologue APRIL has an important role in the selection, maturation and survival of B cells and plays a significant role in the pathogenesis of systemic lupus erythematosus (SLE). Objectives The aim of the study is to evaluate BAFF and APRIL in patients with SLE. Methods The 30 pts (80% female, age 31,0 [28,0-45,0 years] (mediana [interquartile range 25%>75%])) with SLE (ACR criteria, 1982) and 27 controls (93% female without any rheumatic and infectious diseases, age 27,0 [26,0-30,0] years) were examined. We considered SLE-related factors: disease duration, clinical features, SLE Disease Activity Index (SLEDAI 2K), Systemic Lupus International Collaborating Clinics (SLICC) damage index and treatment. All patients were evaluated for laboratory data (ESR, CRP, immunoglobulin G, A and M, complement fragments C3 and C4 and others), autoantibodies (ANA, antiDNA, ENA-SSA, -SSB, -Sm, aPL) and CD19 B lymphocytes subpopulation. The concentration of soluble BAFF and APRIL (ng/ml) were determined in serum samples by ELISA (Bender MedSystem GmbH, Austria). B cells were detected by flow cytometry. Statistical analysis has been performed with STATISTICA program, version 8.0. Results The mean disease duration of SLE was 7,0 [2,0-10,0] years, SLEDAI 2K score - 4 [2-13], SLICC damage index score - 0 [0-1], current prednisone dose – 10,0 [7,5-25,0] mg/day. SLE pts did not differ from healthy controls in BAFF (0,02 [0,02-0,08] vs 0,02 [0,02-0,02] ng/ml) and APRIL (0,01 [0,01-0,62] vs 0,01 [0,01-0,01] ng/ml) levels. Among SLE pts BAFF level correlated with lupus anticoagulant (r=0,88, p<0,05) and ESR (r=0,43, p<0,05); APRIL level correlated with SLEDAI 2K (r=0,55, p<0,01), CRP (r=0,50, p<0,01), antiDNA (r=0,38, p<0,05), creatinine level (r=0,39, p<0,05), current prednisone dose (r=0,55, p<0,01) and CD19 B lymphocytes absolute count (r=0,89, p<0,05). We divided SLE pts on two groups: the 1st- pts with high activity (SLEDAI 2K≥8), the 2nd – pts with low (SLEDAI 2K<8). The patients of 1st group had higher level of APRIL (1,69 [0,01-4,0] vs 0,01 [0,01-0,01] ng/ml, p<0,05) compared to 2nd group and control, there is no difference in BAFF level in these groups. Conclusions In our study there are no differences in BAFF and APRIL levels in patients with SLE and healthy control. This result may be explained by successful therapy and suppression of disease activity in our patients. Patients with high activity of SLE had increased level of APRIL, there is no correlation in disease activity and BAFF level. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.3582
BackgroundThe prevalence of anti-DFS70 antibodies in healthy volunteers (HV) by various methods ranges from 0 to 22% [1]. This pattern, which is rare in patients with systemic autoimmune rheumatic diseases (SARDs), has been described as the second most common in serum HV [2].ObjectivesTo study the frequency of detection of anti-DFS70 in HV, patients (pts) with undifferentiated SARDs and systemic lupus erythematosus (SLE).MethodsA total of 45 HV, 17 undifferentiated SARDs pts and 81 SLE pts were included in the study. Groups were comparable in gender and age among themselves. The diagnosis of SLE was performed according to the ACR/EULAR 2019 classification criteria. Serum samples were tested for anti-DFS70 (ANA HEp-2 ELITE/DFS70 knock-out IFA, “Trinity Biotech”, Ireland). Fluorescence titers ≥1:160 were considered as positive for ANA HEp-2 cell patterns.ResultsPositive results of the ANA study were found in 81 (100.0%) pts with SLE, in 16 (94.0%) pts with undifferentiated SARDs and 7 (15.6%) HV (Table 1). Abs to DFS70 were isolated, detected in 4 (57.1%) HV, in 9 (56.2%) undifferentiated SARDs, but were not detected in SLE pts. Classical HEp-2 cell patterns (homogeneous AC-1, speckled AC-4.5, homogeneous+speckled AC-1,4,5, cytoplasmic AC-19,20,21) without anti-DFS70 were deficient in 3 (42.9%) HV, in 7 (43.8%) patients with undifferentiated SARDs and in 81 (100%) patients with SLE (Table 2). Therefore, monospecific anti-DFS70 were detected in the HV and undifferentiated SARDs groups, but were not detected in patients with a reliable diagnosis of SLE.ConclusionMonospecific anti-DFS70 can potentially be considered as a possible marker for the exclusion of SARDs and a sign of benign autoimmunity.References[1]Conrad K, Röber N, Andrade LEC, Mahler M. The Clinical Relevance of Anti-DFS70 Autoantibodies. Clinic Rev Allerg Immunol (2017) 52:202–216.[2]Mariz HA, Sato EI, Barbosa SH, Rodrigues SH, Dellavance A, Andrade LEC (2011) Pattern on the antinuclear antibody-HEp-2 test is a critical parameter for discriminating antinuclear antibody-positive healthy individuals and patients with autoimmune rheumatic diseases. Arthritis Rheum 63:191–200.Disclosure of InterestsNone declared
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