Anti-Cyclic Citrullinated Peptide Antibodies in Patients with Juvenile Idiopathic Arthritis

The objectives were to determine the prevalence and clinical significance of anti-cyclic citrullinated peptide (anti-CCP) antibodies in patients with juvenile idiopathic arthritis (JIA). Anti-CCP antibodies were checked by ELISA in 68 children with JIA, 38 males and 30 females with mean age of 10.6 (±4.02) years and disease duration of 3.7 (±1.8) years. Thirty-eight (56%) patients had polyarticular disease, 20 (29%) patients had oligoarticular disease and 10 (15%) patients had systemic onset disease. All patients had their antinuclear antibodies (ANA), rheumatoid factor (RF) and ESR checked and x-rays performed to look for erosions. Results were compared to those of 20 healthy children, 14 children with juvenile systemic lupus erythematosus (JSLE) and 30 adults with rheumatoid arthritis (RA). Anti-CCP antibodies were positive in 14/68 (20.6%) patients with JIA, all had polyarticular-onset disease. All patients with positive anti-CCP antibodies had RF-positive polyarthritis. Anti-CCP antibodies were negative in all patients with oligoarticular-onset and systemic-onset disease including 2 patients with extended oligoarthritis. Anti-CCP antibodies were negative in healthy and JSLE controls but were positive in 20/30 (66.5%) adults with RA. Anti-CCP antibodies correlated significantly with joint erosions in patients with JIA (p?=?0.004) but no significant relation was found between anti-CCP antibodies and ANA positivity or raised ESR. These data confirm that anti-CCP antibodies are less prevalent in JIA than adult RA but are detectable in a significant proportion of RF-positive patients with polyarticular-onset JIA. The current study showed significant relation between anti-CCP positivity and erosive joint disease in JIA.     

Antibodies to nucleic acids in systemic lupus erythematosus and rheumatoid arthritis patients and their relatives

Sera of systemic lupus erythematosus (SLE) patients contain antibodies to double-stranded and single-stranded DNA while antibodies found in rheumatoid arthritis sera are specific mainly to single-stranded DNA. Anti-RNA antibodies in the both cases are directed against double-stranded RNA and belong to the IgM class while anti-DNA antibodies are IgG. Genetic variance analysis based on family correlations suggests that the synthesis of antibodies to DNA is subject to different modes of gene regulations in systemic lupus erythematosus and rheumatoid arthritis patients. In the case of SLE, a high degree of genetic determination is demonstrated, due mainly to the X-chromosomal component of the general phenotypic variance. The failures in the immunoregulation system that are responsible for anti-RNA antibody production are rather similar in the two groups of patients and involve a combination of environmental and autosomal factors.     

Antinuclear antibodies in Thai patients with connective tissue diseases

A study of antinuclear antibodies (ANA) among Thai patients with various connective tissue diseases revealed that the prevalence of ANA was similar to that in other countries, but that the ANA patterns showed interesting contrasts in most diseases. Rather than the predominant homogeneous pattern seen elsewhere in systemic lupus erythematosus and rheumatoid arthritis, the speckled pattern was commonest among Thai patients with these two diseases (67.9% and 76.9% respectively). Patients with scleroderma exhibited a much lower percentage of the nucleolar pattern (17%) than reported elsewhere. The discrepancy between our findings and those from other studies may reflect differences in genetics, the environment or the severity of disease.     

A cross-sectional hospital-based study of autoantibody profile and clinical manifestations of systemic lupus erythematosus in south Indian patients

Our study was aimed to analyze clinical manifestations, autoantibodies and other serological abnormalities in South Indian patients with systemic lupus erythematosus. Clinical history and findings on systemic examination were noted. Antinuclear antibodies (ANA), antibodies to double-stranded DNA (dsDNA) were detected by immunofluorescence and ANA profile by Immunoblotting. Arthritis was most common followed by fever and skin rash. Clinical manifestations vary according to geographical location of the patient. ANA was positive in 64.28% and anti-dsDNA in 89.36% of patients. All patients with lupus nephritis were positive for dsDNA. Detection of antibodies to dsDNA, RNP and anti-Smith (Sm) are of diagnostic and prognostic importance.     

Antibodies against nuclear poly(A) polymerases in rheumatic autoimmune diseases

Sera from 53 patients, 26 with systemic lupus erythematosus (SLE), 8 with rheumatoid arthritis (RA), 9 with Sjogren's syndrome (SS), and 10 with scleroderma (Scl), were screened for the presence of antibodies against liver-type poly(A) polymerase and tumor-type poly(A) polymerase. Sixty percent of the patients with the above four autoimmune diseases have antibodies directed against liver poly(A) polymerase, whereas sera from 74% of the patients contained anti-hepatoma poly(A) polymerase antibodies. About 25% of the patients produced antibodies exclusively against the tumor poly(A) polymerase. IgG containing anti-liver or anti-tumor poly(A) polymerase antibodies inhibited the activity of the respective enzyme. IgG containing antibodies against liver and tumor enzymes inhibited the activity of both enzymes, whereas IgG from sera that did not react with poly(A) polymerase had no effect on either enzyme. These data demonstrated the specificity of these autoantibodies and confirmed the results of the radioimmunoassay.     

Vaccination against meningococcus in complement-deficient individuals

Autoantibodies to CRP were reported previously in patients suffering from toxic oil syndrome. This syndrome resembles autoimmune diseases such as systemic lupus erythematosus (SLE) or systemic scleroderma. We therefore examined the prevalence of antibodies to CRP and other acute-phase proteins in autoimmune diseases, including SLE, subacute cutaneous lupus erythematosus (SCLE), systemic scleroderma (SSc), and primary biliary cirrhosis (PBC), as well as in bone marrow transplantation-induced chronic graft-versus-host disease and eosinophilia–myalgia syndrome. IgG antibodies to CRP were found in 78% of SLE and in 30% of SCLE patients, while 16% of patients with PBC were positive. In up to 45% of patients with SSc predominantly IgG antibodies to ceruloplasmin were detectable. Lack of systemic involvement as in discoid lupus erythematosus and localized scleroderma (morphea) correlated with low or absent antibody formation. However, no correlation was found between anti-acute-phase protein antibodies with liver disease or other organ involvement. Adsorption studies revealed that non-native epitopes on the CRP molecule, termed modified CRP, are the main target of antibodies. Chronic inflammatory tissue injury in systemic autoimmune disease might increase the presentation of cryptic epitopes of CRP to the threshold required for T cell activation.     

Increased Prevalence of Anti-Third Generation Cyclic Citrullinated Peptide Antibodies in Patients With Rheumatoid Arthritis and CREST Syndrome

To investigate the prevalence of anti-third generation cyclic citrullinated peptide antibodies (anti-CCP3) in patients with systemic connective tissue diseases, we assembled a training set consisting of 115 patients with rheumatoid arthritis (RA), 52 with Calcinosis, Raynaud’s phenomenon, oesophageal dysmotility, sclerodactyly, telangiectasia (CREST) syndrome, 21 with scleroderma, 20 with ankylosing spondylitis, 18 with reactive arthritis, 25 with juvenile rheumatoid arthritis (RA), 51 with osteoarthritis, 26 with mixed connective tissue disease, 23 with primary Sjogren’s syndrome, 74 with systemic lupus erythematosus, 49 with Polymyaglia rheumatica, and 39 with polymyositis/dermatomyositis. The commercial enzyme-linked immunosorbent assay (ELISA) was used to detect anti-CCP antibodies, including anti-CCP2 (regular, second generation of CCP antigen) and anti-CCP3 (third generation of CCP antigen) in disease-related specimens and normal controls. These serum samples were also evaluated for anti-centomere antibodies by anti-centromere ELISA kit. The higher frequencies of anti-CCP3 and anti-CCP2 were detected in 75.6 and 70.4% patients with RA, respectively. At the same time, anti-CCP3 (not anti-CCP2) was significantly increased in samples isolated from patients with CREST syndrome. The clinical sensitivity of IgG anti-CCP3 for the patients with CREST syndrome was 29% (15 of 52) and the specificity was 96% (384 of 397), with the exception of the RA group. The anti-centromere antibodies were significantly higher in patients with CREST only. The results of our study suggest that compared to anti-CCP2 assay, the new anti-CCP3 assay can enhance the clinical sensitivity for diagnosis of RA and, as an associate marker combined with anti-centromoere, can distinguish CREST syndrome from other systemic connective tissue diseases, especially RA. The clinical specificity of anti-CCP3 was lower than anti-CCP2 assay in diagnosis of RA because of the crossreaction to the patients with CREST syndrome.     

Anti-Idiotypic Antibodies of Reovirus as Biochemical and Immunological Mimics

Among the autoantibodies that are known to play a role in the pathogenesis or autoimmune diseases, antibodies to DNA (anti-DNA) have been the subject of much study. Several interesting observations have resulted. The ability to make antibodies that bind DNA is not abnormal. Normal mice and humans can produce antibodies that bind DNA. On the other hand, large quantities of antibodies to DNA are found in the sera of patients with systemic lupus erythematosus (SLE), and complement-lixing antibodies to double-stranded (ds) DNA cause some of the tissue lesions, especially glomerulonephritis (GN). Why, then, do some individuals make anti-DNA that deposits in glomeruli, skin, and other tissue, resulting in organ damage? It is likely that disease results from a combination of several factors— ability to make pathogenic antibody subsets, inability to downregulate those subsets, and “tissue susceptibility” to injury from those antibodies and their immune complexes. This chapter will focus on the characteristics of pathogenic antibody subsets and their regulation     

Increased presence of common systemic lupus erythematosus (SLE) anti-DNA idiotypes (16/6 Id, 32/15 Id) is induced by procainamide

Sixty-seven patients on treatment with procainamide were examined for the presence of two common idiotypes of anti-DNA antibodies (16/6 Id and 32/15 Id). These idiotypes have been shown previously to have clinical relevance in patients with systemic lupus erythematosus (SLE). An enzyme-linked immunosorbent assay (ELISA) with rabbit anti-Id antibodies revealed increased concentrations of the 16/6 Id and 32/15 Id in 25 (37%) and 16 (24%) patients, respectively. Five of eight patients with drug-induced lupus had elevated titers of both idiotypes. A high correlation (R=0.56,P<0.001 for 16/6 Id) was found between Id levels and anti-single-stranded DNA (ssDNA) antibody titers and between 16/6 Id titers and antihistone antibodies (IgG,R=0.43; IgM,R=0.25). It seems that procainamide, a component known to be associated with drug-induced lupus, may induce an increased production of common anti-DNA idiotypes in apparently normal subjects.     

Cross-reactive idiotypes in anti-DNA antibodies of systemic lupus erythematosus patients

Cross-reactive idiotypes associated with the combining site of anti-double-stranded DNA antibodies from systemic lupus erythematosus patients were demonstrated by the ability of isologous lupus sera to block functionally the binding of target anti-DNA antibodies to DNA in vitro. A framework idiotype, denoted AM Id, was identified using xenogeneic anti-idiotype antibodies rendered specific by affinity absorptions. The AM Id was found in 85% of patients with systemic lupus erythematosus (n = 63) and correlated positively with anti-DNA antibodies. Analysis of the distribution of the AM Id among individuals showed that, while present in anti-DNA antibodies to varying degrees in individual patients, it was also found in variable amounts on non-DNA-binding immunoglobulins. These results indicate that the AM Id and anti-DNA antibodies represent overlapping populations of immunoglobulins.     

Anti-annexin II Antibodies in Systemic Autoimmune Diseases and Antiphospholipid Syndrome

Objectives

The objective of this study were (1) to evaluate the prevalence of anti-annexin II antibodies in patients with various autoimmune diseases and antiphospholipid syndrome and (2) to correlate anti-annexin II antibodies with anti-phospholipid antibodies.

Materials and Methods

Anti-annexin II antibodies and anti-phospholipid were detected, using an enzyme-linked immunosorbent assay, in the serum of patients with primary antiphospholipid syndrome (n?=?16), systemic lupus erythematosus (n?=?53), primary Sjögren syndrome (n?=?71), systemic sclerosis (n?=?17), systemic vasculitis (n?=?18), and rheumatoid arthritis (n?=?119). Healthy blood donors (n?=?99) were used as controls.

Results

Anti-annexin II antibodies were significantly more prevalent in patients with connective tissue diseases (8.5%), especially antiphospholipid syndrome (14.8%) and rheumatoid arthritis (10%), than in controls (2%). An inverse correlation was observed between anti-annexin II antibodies and antiphospholipid antibodies.

Conclusion

Annexin II can be recognized by antibodies in serum from patients with systemic autoimmune disorders. Further studies are required to determine the clinical significance of anti-annexin II antibodies in rheumatoid arthritis and to determine their diagnostic value in discriminating clinical subgroups of patients with antiphospholipid syndrome.     

Tissue specificity of antinuclear antibodies in scleroderma.

Studies of antinuclear antibodies (ANA) were carried out in 39 cases of systemic scleroderma and for comparison in 19 cases of systemic lupus erythematosus (SLE) using indirect immunofluorescence (IF) methods under standard conditions. The results on three different substrates--monkey esophagus, guinea pig lip and rat liver--are reported. In 48.7% of scleroderma cases ANA showed a substrate specificity. The highest percentage of positive results in scleroderma was obtained on monkey esophagus (97.4%) and the lowest on rat liver (61.5%). In SLE, in contrast, only about 13% of the sera displayed such specificity. If only sera with substrate specificity are considered, the positive results on monkey esophagus and rat liver are 94.7% and 21.1%, respectively. Titers of sera reacting positively on 2 or 3 substrates were mostly in agreement, although some sera both in systemic scleroderma and SLE showed higher titers on monkey esophagus. The IF pattern was usually the same regardless of the substrate. Tests for ANA in scleroderma should be performed on at least 2 substrates simultaneously.     

Reactive oxygen species modify human DNA, eliciting a more discriminating antigen for the diagnosis of systemic lupus erythematosus.

During the development of an ELISA to measure anti-DNA antibodies in systemic lupus erythematosus (SLE) sera, native dsDNA was found not to be the most appropriate antigen to use in ELISA assays for differentiating between SLE patients and those with rheumatoid arthritis (RA), a disease also associated with circulating serum anti-DNA antibodies. By modifying the ELISA technique to incorporate human DNA, denatured by reactive oxygen species, to detect anti-DNA antibodies in SLE sera, results consistently showed an increase in antibody binding when compared with the native antigen; no such trend was observed in the comparable group of RA patients. Using this assay serum anti-dsDNA antibody levels were measured in a group of 20 controls, 20 RA patients (10 seropositive and 10 seronegative) and 30 SLE patients (15 with clinically active disease, 15 with inactive disease). A comparison with the standard radioimmunoassay used to measure anti-DNA antibodies for the diagnosis of SLE showed that the ELISA assay using modified DNA performed better than the standard radioimmunoassay offering an improvement in both clinical specificity and sensitivity. The improved method particularly reduced the problem of false-negative results for SLE patients shown clinically to be either mildly active or inactive.     

Antiviral antibodies in rheumatoid synovial fluid and cryoprecipitates

Distribution of viral antibodies was examined by the indirect fluorescent antibody technique in specimens from rheumatoid arthritis and from systemic lupus erythematosus patients. Titres in sera from rheumatoid arthritis patients were usually equal to, or greater than, those in joint fluids. Viral antibodies were found in whole cryoprecipitates from joint fluids and in the IgG fractions of the cryoprecipitates. Anti-herpes simplex antibodies were most frequently found in cryoprecipitates and were at highest titre in sera and joint fluids. Anti-respiratory syncytial virus antibodies were next in frequency and in titre. No evidence was presented to support the concept of localized viral antibody synthesis in joints of rheumatoid arthritis patients. Serum antibody titres in systemic lupus erythematosus patients were similar to those in rheumatoid arthritis patients; however, viral antibodies were not found in the serum cryoprecipitates of systemic lupus erythematosus patients.     

Evaluation of anti-nuclear antibodies and kidney pathology in Lewis rats following exposure to Libby amphibole asbestos

Abstract

The prevalence of anti-nuclear antibodies (ANA) and self-reported systemic autoimmune diseases were increased in residents of Libby, MT, as was the incidence of ANA in Lewis rats exposed to Libby amphibole (LA) asbestos. However, rats induced to develop rheumatoid arthritis (RA) did not develop autoantibodies associated with RA, nor was RA exacerbated by LA exposure, suggesting that increased ANA expression might be related to some other autoimmune process. Libby residents self-reported increased numbers of physician-diagnosed cases of systemic lupus erythematosus (SLE). Thus, the goal of this study was to determine if the increased incidence of ANA in Lewis rats exposed to LA is related to the development of SLE-like disease. Female Lewis rats were intratracheally instilled bi-weekly for 13 weeks with total doses of 0.15, 0.5, 1.5, or 5.0?mg of LA or 0.5 or 1.5?mg of a positive control fiber, amosite. ANA incidence was significantly increased in all asbestos dose groups, although no dose response was observed. The occurrence of proteinuria was increased in LA 0.5, LA 5.0, and amosite 0.5 dose groups; however, the microscopic appearance of the kidneys was normal, no binding of autoimmune complexes to glomerular surfaces was observed, and antibodies to double-stranded DNA were not elevated. Therefore, an increased prevalence of ANA in rats exposed to asbestos does not appear to correlate with disease markers typically observed in SLE. Analysis of ANA specificity for extractable nuclear antigens (ENA) determined that 98% of ENA⁺ samples were specific for the Jo-1 antigen. Autoantibodies to Jo-1 have been reported in patients with interstitial lung disease, suggesting that autoantibodies to Jo-1 may be a biomarker for asbestos-related pulmonary disease.     

Evidence in patients with systemic lupus erythematosus of the presence of antibodies against RNA-dependent DNA polymerase of baboon endogenous virus.

Immunoglobulin G (IgG) in six out of 30 patients with systemic lupus erythematosus (SLE) strongly inhibited the activity of RNA-dependent DNA polymerase (RDPase) of baboon endogenous virus, M7, while IgG obtained from scleroderma patients, rheumatoid arthritis patients and normal subjects was less reactive. Experiments with anti-human IgG and with IgG F (ab')2-bound immunoaffinity columns indicated that the inhibition of RDPase was antibody-mediated. The RDPase inhibiting activity of SLE IgG was considered not to be due to cross-reactions of anti-nuclear antibodies including anti-DNA, anti-ribonucleoprotein, anti-Sm and anti-SS.B antibodies. SLE IgG preferably inhibited the RDPase activity of baboon endogenous virus and a feline endogenous virus, RD114. These findings support the hypothesis that retrovirus(es) might be involved in SLE.     

Antibodies cross-reactive with DNA and cardiolipin in patients with systemic lupus erythematosus

Using a modified solid phase radioimmunoassay (RIA), sera from 11 patients with systemic lupus erythematosus (SLE) that had high DNA binding activities were analysed for their ability to cross-react with phospholipids. Polyspecific anti-DNA antibodies that reacted with both ssDNA and dsDNA had the ability to cross-react with phospholipids, especially cardiolipin. The relationship between biological false-positive serological tests for syphilis (BFP-STS) and the magnitude of serum anti-DNA antibody levels is also discussed.     

Idiotypic mimicry of a cell surface DNA receptor: evidence for anti-DNA antibodies being a subset of anti-anti-DNA receptor antibodies.

Anti-idiotypic anti-DNA antibodies (anti-anti-DNA) have previously been described in both patients with systemic lupus erythematosus and healthy individuals. Jerne's hypothesis predicts that such antibodies would bear a paratope reactive with non-sequence specific DNA binding proteins. Here we have explored the notion of a molecular mimicry between anti-anti-DNA antibodies and antibodies to a previously described 28-29 kD cell surface DNA binding molecule. It was shown that affinity purified anti-anti-DNA antibodies inhibit the binding of DNA to cells and that MoAb to the 28-29 kD receptor react with anti-DNA antibodies. These findings indicate that a subset of anti-anti-DNA antibodies are idiotypically related to antibodies reactive with a cell surface DNA binding molecule. It is hypothesized that anti-DNA antibodies may arise when a convergence of genetic and environmental influences favours an unrestrained anti-idiotypic response to cell surface DNA binding molecule(s).     

Antibodies against Ku protein in sera from patients with autoimmune diseases

Immunoaffinity-purified Ku protein was used to screen sera from patients with systemic lupus erythematosus (SLE), scleroderma, myositis and Sjögren''s syndrome for anti-Ku antibodies in a quantitative immunoblot assay. Sixteen percent of the 159 studied sera were reactive with the Ku protein; significantly increased frequencies of anti-Ku antibodies were found in SLE (19%) and scleroderma (14%) sera. Patients with myositis and Sjögren''s syndrome showed similar frequencies. All positive sera had antibodies to the 86 kD subunit of Ku protein; only one serum did not react with 70 kD subunit. Frequencies of other autoantibodies were compared in anti-Ku positive and negative patients. Only anti-Sm antibodies, especially in the absence of anti-nRNP, appear to be associated with the presence of anti-Ku antibodies. A strong correlation between anti-Ku antibodies and the class II HLA antigen DQw1 (89% of the positive sera) was observed, suggesting participation of MHC genes in the mounting of the anti-Ku immune response.     

Monoclonal human anti-DNA antibodies from EB virus-transformed lymphocytes of systemic lupus erythematosus (SLE) patients

Sixteen monoclonal human anti-DNA antibodies were obtained from Epstein-Barr virus-transformed lymphoblastoid cells of patients with systemic lupus erythematosus (SLE) and were studied in terms of antigenic specificity. All of the antibodies showed polyspecificity to polynucleotides. Among them, some antibodies had a specificity to single-stranded (ss) DNA. Especially, O-8 antibodies showed a preference for polynucleotides with pyrimidine bases. The binding specificity of the antibody was also studied using different sizes of dT oligomers in order to assess the size of the epitope. It was revealed that oligonucleotides with a size of more than 25–30 nucleotides are required for inhibition of the antibody to ss-DNA. Other studies also demonstrated that anti-ss-DNA (O-8) antibody and anti-double-stranded (ds) DNA (NE-28) antibody bound to different combining sites in the same polynucleotides, poly(dT). These results suggest that some anti-ss-DNA antibodies are directed to the conformational structure related to the base sequence and that nucleic acids, therefore, might be responsible for the possible immunogenic stimulus causing the anti-DNA immune response. We also indicate that this type of antibody would be popular among serum anti-DNA antibodies in SLE.