Anti-Ro Positivity in Systemic Lupus Erythematosus: Association with Clinical Phenotypes and Serological Profile

Background: Anti-Ro antibody positivity (anti-Ro+) can have possible association with the clinical features and serology in patients with systemic lupus erythematosus (SLE). Methods: This retrospective observational study enrolled 158 SLE patients at the Rheumatology department, Fauji Foundation Hospital, from 1st May to 31st October 2023. Demographic and clinical data were collected. Patients were stratified by anti-Ro (positive vs negative), concomitant anti-La (anti-Ro+/ anti-La+; anti-Ro- / anti-La-; anti-Ro+/anti-La- and anti-Ro-/anti-La+), and ANA status (positive vs negative). Mann-Whitney U and Chi-square tests were employed for comparisons, with statistical significance set at p[Formula: see text]0.05 Results: Study population consisted mainly of females (98.7%), with median age of 30± 20 years. Anti-Ro antibodies were positive in 74(46.8%) patients. This group had significantly higher discoid rash (16.2% vs 6%, p-value 0.04), sicca symptoms (41.1% vs 31%, p-value 0.005), anemia (71.6% vs 56%, p-value 0.04) compared to anti-Ro negative group. No significant differences were observed in nephritis (29.7% vs 34.5%, p=0.52) or neurological disease (9.5% vs 11.9%, p=0.69) between the two groups. Anti-Ro+ /anti-La+ group had higher myositis (20.6%), sicca symptoms (55.9%) and fever (70.6%) compared to other groups. ANA+/anti-Ro+ group (48.6%) had significant association with sicca symptoms (50.7%) and fever (64.2%). Conclusion: Anti-Ro positivity in SLE is associated with muco-cutaneous manifestations and anemia. Concomitant anti-La positivity is linked to increased xerophthalmia, myositis, and fever. While anti-Ro status is independent of ANA, the combination of both autoantibodies is associated with increased sicca symptoms and fever in SLE patients.

Atherosclerosis is an inflammatory disease and the major cause of cardiovascular disease (CVD) in general. Atherosclerotic plaques are characterized by the presence of activated immune competent cells, but antigens and underlying mechanisms causing this immune activation are not well defined. During recent years and with improved treatment of acute disease manifestations, it has become clear that the risk of CVD is very high in a prototypic autoimmune disease, systemic lupus erythematosus (SLE). SLE-related CVD and atherosclerosis are important clinical problems but may in addition also shed light on how immune reactions are related to premature atherosclerosis and atherothrombosis. A combination of traditional and nontraditional risk factors, including dyslipidaemia (and to a varying degree hypertension, diabetes and smoking), inflammation, antiphospholipid antibodies (aPL) and lipid oxidation are related to CVD in SLE. Premature atherosclerosis in some form leading to atherothrombosis is likely to be a major underlying mechanism, though distinctive features if any, of SLE-related atherosclerosis when compared with 'normal' atherosclerosis are not clear. One interesting possibility is that factors such as inflammation or aPL make atherosclerotic lesions in autoimmune disease more prone to rupture than in 'normal' atherosclerosis. Whether premature atherosclerosis is a general feature of SLE or only affects a subgroup of patients remains to be demonstrated. Treatment of SLE patients should also include a close monitoring of traditional risk factors for CVD. In addition, attention should also be paid to nontraditional risk factors such as inflammation and SLE-related factors such as aPL. Hopefully novel therapeutic principles will be developed that target the causes of the inflammation and immune reactions present in atherosclerotic lesions.

Objective To examine clusters of anti-nuclear antibodies (ANA) and their associations with clinical features in patients with systemic lupus erythematosus (SLE). Methods It was a retrospective study. 113 SLE patients were reviewed from March 2010 to May 2012 in Department of Rheumatology, Jinhua Central Hospital. ANA and specific autoantibodies to 15 kinds of nuclear antigens were tested by indirect immunofluorescence assay (IIF) and line immunoassay (LIA) respectively. Hierarchical clustering method was performed to analyze specific clusters of ANA profiles in SLE. Chi-square tests were used to investigate relationship between antibody clusters and clinical features of SLE. Results The positive rate of LIA for ANA was 97.3%,consistent with IIF method, and the total accordance rate of the both methods was 98.2%. Thirteen kinds of antigen-specific antibodies were detected in SLE patients by LIA. Clustering analysis for these antibodies showed three specific clusters in SLE, Nuc/His/dsDNA cluster (C1), low-Ro/low-La cluster (C2), and Ro/Sm/RNP cluster (C3), accounting for 36.3%, 24.8%, 38.9% of the total cases respectively. There were significant difference of AST levels among three clusters ［(32.62±21.92)U/L, (25.56±16.63)U/L, (50.41±60.86)U/L respectively for C1, C2 and C3］. High incidences of chronic cutaneous lupus, abnormal renal indicators and inflammatory synovitis were found in all three clusters. Besides, there were significant differences among three clusters for the incidences of chronic cutaneous lupus (39.0%, 39.3%, 63.6% respectively for C1, C2, C3) and leukopenia/lymphopenia (56.1%, 25.0%, 56.8% respectively for C1, C2, C3) (P<0.05). Patients in Ro/Sm/RNP cluster showed higher incidences of lupus nephritis (43.2%/26.8% or 39.3%); patients in low-Ro/low-La cluster showed low risk of hypertension (7.1%/19.5% or 22.7%); patients in Nuc/His/dsDNA cluster showed high incidences of thrombocytopenia (41.5%/21.4% or 25.0%) and high risk of lung or upper respiratory tract infection (46.3%/28.6% or 29.5%), but low incidence of neurologic symptoms (0%/3.6%% or 11.4%). Conclusion Three characterized ANA clusters are identified in SLE patients in this pilot study. Different clusters are associated with certain clinical features and complications of SLE. However, the correlations found in this study need to be investigated further in larger populations.（Chin J Lab Med,2014,37:587-592） Key words: Lupus erythematosus, systemic; Antibodies, antinuclear; Cluster analysis

Human γδ T cells recognize conserved endogenous and stress-induced antigens typically associated with autoimmune diseases. However, the role of γδ T cells in autoimmune diseases is not clear. Few autoimmune disease-related antigens recognized by T cell receptor (TCR) γδ have been defined. In this study, we compared Vδ2 TCR complementarity-determining region 3 (CDR3) between systemic lupus erythematosus (SLE) patients and healthy donors. Results show that CDR3 length distribution differed significantly and displayed oligoclonal characteristics in SLE patients when compared with healthy donors. We found no difference in the frequency of Jδ gene fragment usage between these two groups. According to the dominant CDR3δ sequences in SLE patients, synthesized SL2 peptides specifically bound to human renal proximal tubular epithelial cell line HK-2; SL2-Vm, a mutant V sequence of SL2, did not bind. We identified the putative protein ligand chaperonin-containing T-complex protein 1 subunit ζ (CCT6A) using SL2 as a probe in HK-2 cell protein extracts by affinity chromatography and liquid chromatography-electrospray ionization-tandem mass spectrometry analysis. We found CCT6A expression on the surface of HK-2 cells. Cytotoxicity of only Vδ2 γδ T cells to HK-2 cells was blocked by anti-CCT6A antibody. Finally, we note that CCT6A concentration was significantly increased in plasma of SLE and rheumatoid arthritis patients. These data suggest that CCT6A is a novel autoantigen recognized by Vδ2 γδ T cells, which deepens our understanding of mechanisms in autoimmune diseases.

Dysregulation of the antiviral immune response may contribute to autoimmune diseases. Here, we hypothesized that altered expression or function of MAVS, a key molecule downstream of the viral sensors RIG-I and MDA-5, may impair antiviral cell signalling and thereby influence the risk for systemic lupus erythematosus (SLE), the prototype autoimmune disease. We used molecular techniques to screen non-synonymous single nucleotide polymorphisms (SNPs) in the MAVS gene for functional significance in human cell lines and identified one critical loss-of-function variant (C79F, rs11905552). This SNP substantially reduced expression of type I interferon (IFN) and other proinflammatory mediators and was found almost exclusively in the African-American population. Importantly, in African-American SLE patients, the C79F allele was associated with low type I IFN production and absence of anti-RNA-binding protein autoantibodies. These serologic associations were not related to a distinct, functionally neutral, MAVS SNP Q198K. Hence, this is the first demonstration that an uncommon genetic variant in the MAVS gene has a functional impact upon the anti-viral IFN pathway in vivo in humans and is associated with a novel sub-phenotype in SLE. This study demonstrates the utility of functional data in selecting rare variants for genetic association studies, allowing for fewer comparisons requiring statistical correction and for alternate lines of evidence implicating the particular variant in disease.

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease associated with significant morbidity and mortality. Sicca symptoms are frequent in SLE which may be related to concomitant occurrence of Sjogren's syndrome (SS). The aims of study were to determine prevalence of oral manifestations and tempromandibular joint disorders, and to find a correlation between the changes in saliva flow rate, pH and composition with the incidence of dental caries in SLE patients. Subjects, materials and methods: One hundred and two individuals were enrolled in this study; 52 of them were SLE patients; and 50 were healthy control individuals matched in age and gender. The assessment of dental status was made according to the decay missing filling teeth (DMFT) index; the gingival inflammation was assessed using the criteria of gingival index; Clinical pocket depth was measured with periodontal probe type William, and whole unstimulated saliva samples have been collected from each subject for biochemical analysis. Also, salivary flow rate and pH were measured. After centrifugation, the supernatant of saliva was aspirated for biochemical analysis. Results: Oral ulceration was the most prominent orofacial manifestations of SLE patients followed by Tempromandibular joint (TMJ) disorders and facial skin rash then oral vesicles& bullae, oral lichen planus and finally oral petechiae & purpura. Salivary flow rate and salivary pH were significantly lower in SLE patients than in the control subjects. Oral hygiene index (DMFT index, gingival index, Clinical pocket depth) were significantly higher in SLE patients than in the control subjects .Salivary calcium, sodium, chloride, and total protein were significantly higher among SLE patients than in the control subjects. While salivary potassium and inorganic phosphorus were significantly lower among SLE patients than in the control subjects. In addition, there was a highly significant positive linear correlation between age of SLE patients and DMFT, and between age and clinical pocket depth; and a highly significant negative linear correlation between salivary flow rate and salivary calcium in SLE patients. Also there was highly significant positive linear correlation between DMFT and salivary calcium, and between DMFT and salivary chloride. Conclusions: Oral manifestations are common in Iraqi SLE patients. Changes in salivary flow rate, pH, salivary composition, and increased dental caries may serve as potential markers of the extent of autoimmune mediated salivary gland dysfunction which is similar to Sjogren's syndrome.

The objective of this study was to perform a longitudinal follow-up of antinuclear antibodies (ANAs) and anticardiolipin antibodies titers (aCL) throughout pregnancy in a group of patients with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) patients during their therapy for the prevention of fetal loss and to examine their relationship with pregnancy outcome. Thirty patients and 15 controls were followed in the study. Fifteen patients had SLE (group I) and 15 had APS (group II, of which seven patients had primary APS and eight had APS secondary to SLE). All patients were receiving therapy for the prevention of fetal loss with prednisone and aspirin as part of an ongoing clinical trial in lupus pregnancy. If there was a history of previous thrombosis, heparin was added. Blood samples were taken at the 1st, 2nd, and 3rd trimesters (T) of pregnancy in order to assess the presence of IgG and IgM aCL antibodies (ELISA), anti-dsDNA ( C. luciliae) ANAs (HEp-2 cells), and immunospecific antibodies (antiextractable nuclear antigens). We collected 90 samples from patients and 45 samples from healthy controls. Group I (SLE) ANAs were positive in 100% during the 1st T, 67% in the 2nd T, and 67% in the 3rd T, with various immunofluorescence patterns. In five patients, aCL antibodies were detected without a history of APS (one in 1st T, three in 2nd T, and one in 3rd T). Fetal loss was observed in two patients, in one of whom it was associated with nephritis, high titers of ANAs, and anti-dsDNA. Another patient had pulmonary hemorrhage with anti-dsDNA and aCL. In group II, all but one patient with primary APS were negative to ANAs. In secondary APS, by contrast, 6/8 patients (75%) had positive ANAs at least during the 1st T. All seven patients with primary APS and 6/8 with secondary APS had aCL during pregnancy. In 9/15 (60%) patients from the APS group with a history of previous fetal loss, aCL became negative during pregnancy and they had live births. The disappearance of aCL was associated with improved fetal survival (relative risk, or RR, 0.67). ANAs in the control group were positive in 7% at low titers, and all of them were negative for aCL. Despite treatment, ANAs are prevalent during pregnancy in SLE patients and APS secondary to SLE. During pregnancy in SLE, aCL titers may appear. Decreasing titers and/or disappearance of aCL correlated with improved fetal prognosis in a subset of patients with APS.

Background:Systemic lupus erythematosus (SLE) is a chronic and potentially severe disease, that may affect virtually every organ and system. The 2019 EULAR/ACR criteria exclude both liver and gastrointestinal involvement. However,...

Identification of HnRNP-A2/B1 as a Target Antigen of Anti-Endothelial Cell IgA Antibody in Behçet's Disease

Belimumab neutralizes the soluble form of B-cell activating factor and is FDA-approved for treating Systemic Lupus Erythematosus (SLE). In this retrospective analysis 16 female SLE patients had been treated with belimumab according to FDA guidelines along with any immunosuppressive drugs or antimalarial drugs they had previously been prescribed, in addition to prednisone. Thirteen of 16 patients presented with serologic evidence of positive anti-nuclear antibody titer while several other SLE patients also had clinical evidence of secondary Sjogren’s Syndrome (2°SS), the latter supported by sicca symptoms and positive anti-Sjogren’s-syndrome-related antigen A antibody titer. Changes in the SLE symptoms after belimumab therapy was assessed by the recipient’s responses on the Short Form-36 questionnaire. Ten of the 16 SLE patients reported a decrease in arthralgias or in symptoms of arthritis while fatigue was reduced in 9/16 patients. However, SLE patients with sicca symptoms failed to show any change in the Short Form-36 score after treatment with belimumab. In conclusion, although the results in this small cohort study supported the use of belimumab as an adjunctive therapy for SLE, sicca symptoms associated with 2°SS did not appear to respond to belimumab.

Alleles of IRF8 are associated with susceptibility to both systemic lupus erythematosus (SLE) and multiple sclerosis (MS). While high type I interferon (IFN) is thought to be causal in SLE, type I IFN is used as a therapy in MS. We investigated whether IRF8 alleles were associated with type I IFN levels or serologic profiles in SLE and MS. Alleles which have been previously associated with SLE or MS were genotyped in SLE and MS patients. The MS-associated rs17445836G allele was associated with anti-dsDNA autoantibodies in SLE patients (meta-analysis OR=1.92). The same allele was associated with decreased serum IFN activity in SLE patients with anti-dsDNA antibodies, and with decreased type I IFN-induced gene expression in PBMC from anti-dsDNA negative SLE patients. In secondary progressive MS patients, rs17445836G was associated with decreased serum type I IFN. Rs17445836G was associated with increased IRF8 expression in SLE patient B cells. In summary, IRF8 rs17445836G is associated with human autoimmune disease characterized by low type I IFN levels, and this may have pharmacogenetic relevance as type I IFN is modulated in SLE and MS. The association with autoantibodies and increased IRF8 expression in B cells supports a role for rs17445836G in humoral tolerance.

The results of studies on association between FCGR2A-R/H131 polymorphism and susceptibility to systemic lupus erythematosus (SLE) in the Asian population are controversial. To derive a more precise estimation on the genetic risk of FCGR2A-R/H131 variant for SLE in Asians, a meta-analysis was performed for genotypes R/R+R/H (dominant effect), R/R (recessive effect), and R allele in fixed/random effects models. Twenty studies involving 4,832 SLE patients and 6,190 controls were included. The meta-analysis showed that FCGR2A-R/H131 variant was associated with development of SLE in overall Asians both at allele or genotype level (R vs H, OR 1.201, 95% CI 1.098-1.315; RR+RH vs HH, OR 1.369, 95% CI 1.115-1.682; RR vs RH+HH, OR 1.305, 95% CI 1.029-1.654). After stratification by ethnic, a significant association of R allele with susceptibility to SLE was observed in the Chinese population (R vs H, OR 1.104, 95% CI 1.030-1.183). Evidence from subgroup analyses in non-Chinese populations (all Asians excluding Chinese population) also showed a significant association (R vs H, OR 1.354, 95% CI 1.207-1.519; RR+RH vs HH, OR 1.705, 95% CI 1.234-2.355). In addition, FCGR2A-R131 allele was associated with a 1.186-fold (95% CI 1.043-1.349) greater risk for the occurrence of nephritis in the Asians population with SLE. After stratification by ethnic, this significant association was only consistently identified in non-Chinese populations (R vs H, OR 1.220, 95% CI 1.002-1.486). In summary, FCGR2A-R/H131 polymorphism is associated with SLE and lupus nephritis in Asians.

To evaluate the production of interleukin-10 (IL-10) as well as levels of IgG and antinuclear antibodies (ANA) in systemic lupus erythematosus (SLE) patients and their first-degree relatives and spouses in Icelandic SLE multicase families. IL-10 production was studied by enzyme-linked immunospot assay of freshly isolated peripheral blood mononuclear cells. Total IgG and ANA were also investigated. Subjects consisted of 23 SLE patients and 47 of their first-degree relatives in 9 Icelandic multicase families. Subjects were ethnically matched by a group of healthy controls. A separate study investigated 12 SLE patients (also from SLE multicase families) and their spouses and a matched group of healthy controls. A predefined protocol was used to obtain both clinical and laboratory data, including information about SLE and other autoimmune disorders. The SLE patients had a significantly higher number of IL-10-producing cells compared with both first-degree relatives and healthy controls (P = 0.0005 and P < 0.0001, respectively). First-degree relatives also had a significantly higher number of IL-10-producing cells compared with healthy controls (P = 0.01). This was also true for the spouses of SLE patients, who had a higher number of IL-10-producing cells compared with matched healthy controls (P = 0.02). SLE patients and their first-degree relatives, as well as a limited number of healthy spouses of SLE patients, had increased numbers of spontaneous IL-10-producing cells. These data support the hypothesis that IL-10 production may be genetically determined, and may predispose one toward development of SLE. This has previously been suggested by studies of SLE patients and their relatives in another ethnic population, using another method for measuring IL-10 production. Although these data are based on a small number of observations, they suggest that not only genetic but also environmental factors may be of importance in determining IL-10 production, since the spouses of SLE patients also had an increased number of IL-10-producing cells.

IgG antinuclear antibodies (ANAs) are elevated in patients with systemic lupus erythematosus (SLE) compared with patients with discoid lupus erythematosus (DLE). To provide an expanded immunologic view of circulating ANAs in lupus patients, we compared the expressions of IgG, IgM, and IgA ANAs in DLE and SLE patients. In this cross-sectional study, sera from age-, gender-, and ethnic-matched SLE (N = 35), DLE (N = 23), and normal patients (N = 22) were tested for IgG, IgM, and IgA ANAs using enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) with monkey esophagus as substrate. ELISAs showed elevated levels of IgG ANA, IgM ANA, and IgG/IgM ANA ratios in SLE patients compared with DLE and normal patients. IgA ANA expression was higher in SLE and DLE patients versus normal patients. IIF studies showed higher percentages of patients positive for IgG, IgM, and IgA ANAs in the SLE group. Higher IgG/IgM ANA ratios in SLE than DLE show enhanced class-switching and a more sustained humoral response in SLE. They also suggest a potential connection of IgM ANAs with disease containment.

Nonstandard and adjunctive medical therapies for systemic lupus erythematosus