Gabriella Morozzi

HLA-DPB1 alleles association of anticardiolipin and anti-beta2GPI antibodies in a large series of European patients with systemic lupus erythematosus.

Our objective was to determine the HLA-DPB1 allele associations of anticardiolipin (aCL) and anti-beta2 GPI (aβ2 GPI) antibodies, and of clinical manifestations of the antiphospholipid syndrome (APS), in systemic lupus erythematosus(SLE). We studied 577 European patients with SLE. aCL and aβ2 GPI antibodies were measured by ELISA. Molecular typing of HLA-DPB1 locus was performed by polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. aCL showed positive association with -DPB1*1501 (P 0.005, OR 7.4), and -DPB1*2301 (P 0.009, OR 3.3). aβ2 GPI showed positive association with -DPB1*0301 (P 0.01, OR 1.9), and -DPB1*1901 (P 0.004, OR 8.1). In addition, livedo reticularis was associated with -DPB1*1401, and Raynaud’s phenomenon with -DPB1*2001. In conclusion, HLA-DPB1 locus may contribute to the genetic predisposition to develop antiphospholipid antibodies and clinical manifestations of the APS in patients with SLE.

Anticardiolipin and anti-β2GPI antibodies in a large series of European patients with systemic lupus erythematosus

Objective. To test the prevalences and the clinical associations of anticardiolipin (aCL) and anti-β2GPI (aβ2GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). Methods. 574 SLE patients from 7 European countries were tested for aCL and aβ2GPI by ELISA methods. Results. aCL of IgG, IgM, and IgA isotypes were detected in 22.8%, 14%, and 13.9% of the patients, respectively. IgG and IgM aβ2GPI were detected in 20% of the patients. The presence of aCL was highly associated with the presence of aβ2GPI. Medium-high titer IgG aCL and aβ2GPI were associated with thrombosis, with similar sensitivity, specificity, and positive predictive value. When present at mediumhigh titer, IgG aCL were associated with thrombocytopenia, IgM aCL with hemolytic anemia, and cerebrovascular accidents, IgA aCL with livedo reticularis and Raynauds phenomenon. Conclusion. aCL, when present at medium-high titer, are as important as ab2GPI, as a risk factor for thrombosis. Medium-high titer a...

The Immunogenetics of the Antiphospholipid Syndrome, Anticardiolipin Antibodies, and Lupus Anticoagulant

Whether a genetic predisposition to develop the antiphospholipid syndrome (APS) and to produce anticardiolipin antibodies (aCL) and lupus anticoagulant (LAC) exists has been addressed by family studies and by population studies on primary APS and on aCL in diseases other than primary APS. Various studies suggest a familial occurrence of aCL and LAC, with or without clinical evidence of APS. This familial tendency could be genetically determined, because APS, aCL, and LAC occur in families carrying haplotypes which contain HLA-DR4, -DR7, and -DRw53. Population studies on primary APS also indicate that HLA genes have a role in conferring susceptibility to develop primary APS. Again, DR4, DR7, and DRw53 are the relevant loci. Population studies on aCL in diseases other than primary APS indicate that aCL are associated with DR4, DR7, and DRw53, at least when they are found in patients with systemic lupus erythematosus. Because HLA-DR4, -DR7, and -DRw53 are in linkage disequilibrium, the genetic association of aCL could be with DRw53 and, depending on the regional frequency of DR4 or DR7, it could be linked with either DR4 or DR7. HLA-DR4 seems to be more important in Anglo-Saxons, whereas DR7 emerges in populations of Latin origin. In this report we review our studies and the pertinent literature in this field.

Dosage and characterization of circulating DNA: present usage and possible applications in systemic autoimmune disorders

The discovery of extracellular nucleic acids in the circulation was firstly reported in 1948. In the last few years it has been demonstrated that the entire spectrum of genetic changes seen in primary tumors could also be detected in the serum of patients with solid tumors. This observation has also opened up exciting possibilities for tumor detection and monitoring. More recently investigators started looking for other forms of non-host DNA in the plasma/serum so that in 1997 the presence of fetal DNA in the plasma/serum of pregnant women was demonstrated. This finding suggested that maternal plasma fetal DNA would be a very valuable material for noninvasive prenatal diagnosis and monitoring. It has been also postulated that the presence of the two-way trafficking of nucleated cells and free DNA between the mother and fetus may have potential implications for the development of certain autoimmune diseases. Concerning autoimmune disorders, Tan was the first author to describe the presence of high levels of circulating DNA in patients with systemic lupus erythematosus (SLE) in 1986. Later on different authors demonstrated that elevated levels of serum DNA was also present in patients with other diseases including rheumatoid arthritis. We have analyzed both circulating free DNA and DNA extracted from nucleated blood cells in scleroderma and in lupus patients but, by using gel electrophoresis, we were able to define the pattern of the DNA, instead of simply dosing its amount in the circulation. We have found that SLE and SSc have anomalous patterns of DNA both in serum and in the Buffy-coat and that these patterns are typical for each disorder. It is possible that understanding the biological significance of the diversity in DNA pattern exhibition in white blood cells may give new insights into the pathophysiology of autoimmune disorders. It is also conceivable that circulating and immune-competent cellular DNA markers might offer the promise of precise quantitative analysis useful for diagnostic purposes, without the need to establish difficult cutoffs as is necessary for protein markers.

Diagnostic value of anti-mutated citrullinated vimentin in comparison to anti-cyclic citrullinated peptide and anti-viral citrullinated peptide 2 antibodies in rheumatoid arthritis: An Italian multicentric study and review of the literature

In the last years, the detection of antibodies (Abs) against citrullinated peptides (ACPA) has largely replaced rheumatoid factor (RF) as the most helpful biomarker in the diagnosis of rheumatoid arthritis (RA). Current assays detect ACPA reactivity with epitopes on various different citrullinated proteins. Among these, anti-cyclic citrullinated peptide (CCP) Abs have been widely demonstrated to be an important diagnostic and prognostic tool because of their high specificity. Recently, citrullinated vimentin, a protein highly released in synovial microenvironment, has been identified as potential autoantigen in the pathophysiology of RA and an enzyme-linked immunosorbent assay (ELISA) for the detection of Abs directed against a mutated citrullinated vimentin (anti-MCV) was developed. Several recent studies evaluating the characteristics of anti-MCV in comparison to anti-CCP Abs, have given conflicting results. Anti-MCV have been demonstrated to perform better than anti-CCP as predictor of radiographic damage. Conversely, its additional diagnostic and prognostic role in comparison to anti-CCP in both early and established RA is controversial. Aim of this study was to evaluate the diagnostic performance of anti-MCV in RA and to compare it to anti-CCP and the recently developed assay targeting viral citrullinated peptide 2 (VCP2) in a large cohort of RA patients (n=285), healthy subjects and other disease controls (n=227). Anti-MCV resulted to have a sensitivity of 59% and a specificity of 92%. In comparison, anti-CCP and anti-VCP2 displayed a sensitivity of 77% and 61% and a specificity of 96% and 95%, respectively. Of interest, at the manufacturer recommended cutoff value of 20U/mL, a high percentage of healthy subjects as well as Epstein Barr (EBV) and hepatitis C (HCV) virus infected patients resulted anti-MCV positive. In our large cohort of RA patients, anti-MCV demonstrated lower sensitivity than anti-CCP and VCP2 test, thus not allowing to confirm previously published data. Moreover, the high rate of detection in infectious diseases limits its diagnostic value in undifferentiated arthritis.

Anticardiolipin and anti-β2GPI antibodies in a large series of European patients with systemic lupus erythematosus : Prevalence and clinical associations

Objective: To test the prevalences and the clinical associations of anticardiolipin (aCL) and anti-β 2 GPI (aβ 2 GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). Methods: 574 SLE patients from 7 European countries were tested for aCL and aβ 2 GPI by ELISA methods. Results: aCL of IgG, IgM, and IgA isotypes were detected in 22.8%, 14%, and 13.9% of the patients, respectively. IgG and IgM aβ 2 GPI were detected in 20% of the patients. The presence of aCL was highly associated with the presence of aβ 2 GPI. Medium-high titer IgG aCL and aβ 2 GPI were associated with thrombosis, with similar sensitivity, specificity, and positive predictive value. When present at medium-high titer, IgG aCL were associated with thrombocytopenia, IgM aCL with hemolytic anemia, and cerebrovascular accidents, IgA aCL with livedo reticularis and Raynauds phenomenon. Conclusions: aCL, when present at medium-high titer, are as important as aβ 2 GPI, as a risk factor for thrombosis. Medium-high titer aCL, but not aβ 2 GPI, are associated with other clinical features of the antiphospholipid syndrome.

Safety of cyclosporin A in HCV-infected patients: experience with cyclosporin A in patients affected by rheumatological disorders and concomitant HCV infection.

Abstract:  Because of the relatively high prevalence of both hepatitis C virus (HCV) infection and autoimmune disorders (ADs), it is not rare to encounter in daily clinical practice patients with ADs also carrying HCV. Corticosteroids and/or immunosuppressant drugs are needed to treat ADs, but they place HCV‐infected patients at risk of worsening the infection. So, rheumatologists have often refrained from using corticosteroids or immunosuppressants in AD when HCV–RNA is also present. Cyclosporin A (CsA) is an immunosuppressive agent used to treat a wide range of ADs, but there is a large evidences in the literature, both in vitro and in vivo, suggesting that CsA also exerts an inhibitory effect on HCV replication at standard therapeutic dose. Therefore, this evidence has opened new ways to improve the therapy and the prognosis in patients with HCV‐related liver diseases, including those with transplants. Recent reports, although limited in number, also suggest the safety of CsA in the treatment of patients with AD and concomitant HCV infection. In this review we also report our personal experience on the combination treatment with CsA and anti‐TNF‐α agents in rheumatoid arthritis.

Antibodies to the lens and cornea in anti-DFS70-positive subjects.

Abstract:  Autoantibodies against DFS70 (dense fine speckles 70) antigen have recently been identified among antinuclear antibodies (ANA) in patients with various inflammatory diseases and in patients with different types of cancer. These antibodies are recognized using indirect immunofluorescence (IIF) on HEp‐2 cells, by a fine speckled nuclear staining in interphase HEp‐2 cells and a positive reaction in the chromosome region of mitotic cells. Given that the DFS70 protein is also known as the lens epithelium‐derived growth factor, this study was performed with two objectives: (a) to assess the prevalence of these antibodies in patients sent for ANA testing and in 334 patients with different types of neoplasia and (b) to determine whether the lens tissue was a suitable substrate for the detection of antibodies specific to lens proteins. During routine workup for ANA detection by the IIF method, we found 172 DFS70‐positive sera among 21,516 consecutive samples (prevalence, 0.8%). In the group of patients with neoplastic disease, 6 of 334 (1.8%) were anti‐DFS70‐positive. DFS70‐positive sera were then assayed by the IIF method on cryostatic sections of mouse eye at a dilution of 1:40 with an anti‐human IgG conjugate. Among the 172 DFS70‐positive samples detected by the ANA screening, 32 (19%) were strongly reactive against the reticular fibers of the lens; 8 (5%) were positive only to the corneal epithelium (nuclear negative); 5 (3%) were positive both for the cornea and the lens fibers; 13 (7%) stained only the nuclei of lens and cornea cells, and 4 (2%) were positive against the ciliary muscle. Among the patients with neoplastic diseases, only one with lung cancer reacted weakly with the reticular fibers of the lens. Sera from 20 healthy blood donors were negative. In this preliminary study, we have shown that the prevalence of anti‐DFS70 antibodies is much lower than previously reported, both in patients screened for ANA and in patients with cancer. We have also seen that some DFS70‐positive sera have antibodies that recognize antigens of the lens. Further studies are needed to investigate the fine specificity and the possible significance of these new autoantibodies.

Cartilage oligomeric matrix protein level in rheumatic diseases: potential use as a marker for measuring articular cartilage damage and/or the therapeutic efficacy of treatments.

Abstract:  Cartilage oligomeric matrix protein (COMP) is a tissue‐specific noncollagenous protein that was first detected in the serum and the synovial fluid of patients suffering from rheumatic disorders, such as rheumatoid arthritis, reactive arthritis, juvenile chronic arthritis, and osteoarthritis. In this review, the authors consider serum COMP levels in different diseases and discuss their study of patients with rheumatoid arthritis treated with anti‐TNF‐α, to evaluate whether COMP is able to predict a rapid and sustained clinical response to these drugs. They observe that patients with high COMP levels have a lower ACR 70 response independently of the state of systemic inflammation, and conclude that COMP seems to have a pathogenetic role that is independent of the mechanisms regulating inflammatory processes.

Evaluation of current methods for the measurement of serum anti double-stranded DNA antibodies.

Abstract:  Autoantibodies to double‐stranded DNA (dsDNA) are, by definition, serological markers of systemic lupus erythematosus. However, the clinical value of anti‐dsDNA antibodies largely depends on the assay principle and analytical variables of the methods used to quantitate and immunologically characterize them. In the present article, an overview of current methods for anti‐dsDNA antibody detection is presented, together with a look at the future trends in technologies newly employed in this field.