Anticentromere antibody induced by immunization with centromere protein and Freund’s complete adjuvant may interfere with mouse early-stage embryo

Background Anticentromere antibody (ACA) is a member of the antinuclear antibody spectrum (ANAs) which has been speculated to be associated with subfertility. Thus, the present study aimed to investigate the induction of ACA production and its potential interference with early-stage embryos. Methods Recombinant centromere protein-A (CENP-A) or centromere protein-B (CENP-B) and complete Freund’s adjuvant (CFA) were used to immunize mice. Serum ACA level was then evaluated by using an indirect immunofluorescence test. Immunofluorescence assay was performed to detect IgG in follicles in ovarian tissues and early-stage embryos. Results Following treatment, serum positive ACA was observed in mice treated with CENP and CFA. Furthermore, IgG were detected in follicular fluid and early-stage embryos from mice treated with CENP and CFA. Conclusions This study preliminarily indicated that ACA induced by CENP and CFA may penetrate into the living embryos of early-stage in mice.

Autoinflammatory/Autoimmunity Syndrome Induced By Adjuvants (ASIA) Due to Silicone Incompatibility Syndrome

The adjuvant-induced autoimmune syndrome (ASIA) is associated with a dysregulation of the innate and adaptive immune system after exposure to chemical compounds, including liquid paraffin, silicone gel, acrylamides, and hyaluronic acid. Due the increase of the use of these compounds in cosmetic procedures, the prevalence of this syndrome is increasing. We present the first report in Ecuador associated to ASIA after an elective silicone breast prosthesis procedure, manifested as polyarthralgia, positive antinuclear antibody, anticentromere antibody, and a moderate positive Sclero-70.

POS0717 AUTOIMMUNE LIVER DISEASE IN ANTICENTROMERE ANTIBODY POSITIVE PRIMARY SJOGREN’S SYNDROME

Background:studies have shown that anticentromere antibody (ACA) positivity in primary Sjogren’s syndrome (pSS) is associated with autoimmune liver diseases, most often primary biliary cholangitis (PBC) and autoimmune hepatitis (AIH) [1, 2, 3], but detailed characteristics of the frequency and severity of liver disease in these patients is not presented in the literature.Objectives:to identify the frequency, structure and characterize the course of autoimmune liver diseases in pSS+ACA.Methods:we observe 82 patients with pSS+ACA. The diagnosis of pSS was established on the basis of Russian 2001 criteria, SSc was excluded based on the ACR/EULAR 2013 criteria [4]. 18 of 82 patients (22%) had a persistent increase in alkaline phosphatase, 11 of them were positive for antimitochondrial antibodies (AMA) and, according to the recommendations of the American Association for the Study of Liver Diseases [5], they were diagnosed with PBC. 7 of 18 patients were AMA-negative, 2 of them had a liver biopsy and the diagnosis of AMA-negative PBC was confirmed, 4 patients who did not have a liver biopsy and 1 patient with hepatitis B were excluded from the study. Also, in 6 of 64 patients without signs of liver damage, an increase in AMA was detected, in 1 of them a liver biopsy was performed and the diagnosis of PBC was confirmed. Thus, the group of patients with pSS+ACA and autoimmune liver diseases included 19 patients: 12 patients with AMA-positive PBC, 2 patients with AMA-negative PBC, and 5 patients with asymptomatic AMA positivity.Results:The median follow-up for 19 patients with pSS+ACA and autoimmune liver diseases was 4 years. AMA were detected in 89.5% of patients, an increase in IgM - in 42.1%, an increase in ALT / AST - 63.2%, a decrease in albumin, prothrombin index and cytopenia - 15.8% (were associated with the development of liver cirrhosis). In most cases, the clinical course of liver disease was characterized by an asymptomatic, slowly progressing course, with no signs of progression during observation. Cirrhosis and portal hypertension were detected in 15.8% of patients, hepatic encephalopathy - in 10.5%. Liver biopsy was performed in 9 patients, PBC was diagnosed in all cases (overlap syndrome with AIH was established in 3 cases). Assessment of PBC histological stages showed signs of stage 1 in 5 patients, stage 2 in 1 patient, stage 3 in 3 patients. Observation of 5 patients with stage 1 PBC and 5 AMA-positive patients without signs of liver damage (median follow-up was 2 years), showed the absence of clinical, laboratory and instrumental progression of liver disease, which is why we believe that these patients have epithelitis of the biliary ducts as manifestation of glandular lesions in pSS, but not PBC.Conclusion:autoimmune liver diseases in pSS+ACA are detected in 23.2% of patients, most of whom develop PBC and epitheliitis of the biliary ducts with the same frequency, less often overlap syndrome of PBC and AIH, and characterized by a mild, slowly progressing course and rarely lead to liver cirrhosis.References:[1]Masako Kita et al. Abnormal Liver Function in Patients with Sjogren’s Syndrome. Acta Med. Nagasaki 41: 31-37.[2]Baldini, Chiara et al. “Overlap of ACA-positive systemic sclerosis and Sjögren’s syndrome: a distinct clinical entity with mild organ involvement but at high risk of lymphoma.” Clinical and experimental rheumatology vol. 31,2 (2013): 272-80.[3]Bournia, Vasiliki-Kalliopi K et al. “Anticentromere antibody positive Sjögren’s Syndrome: a retrospective descriptive analysis.” Arthritis research & therapy vol. 12,2 (2010): R47. doi:10.1186/ar2958.[4]van den Hoogen, Frank et al. “2013 classification criteria for systemic sclerosis: an American College of Rheumatology/European League against Rheumatism collaborative initiative.” Arthritis and rheumatism vol. 65,11 (2013): 2737-47. doi:10.1002/art.38098.[5]Lindor, Keith D et al. “Primary Biliary Cholangitis: 2018 Practice Guidance from the American Association for the Study of Liver Diseases.” Hepatology (Baltimore, Md.) vol. 69,1 (2019): 394-419. doi:10.1002/hep.30145.Disclosure of Interests:None declared

Anticentromere antibody induced by immunization with centromere protein a and Freund’s complete adjuvant may interfere with mouse oocyte meiosis

Background Anticentromere antibody (ACA) is a member of the antinuclear antibody (ANA) family, and recent studies have found that ACA may be associated with oocyte maturation disorders; however, the possible mechanism behind this phenomenon remains unknown. We conducted this study to investigate whether ACA could penetrate into the living oocytes and interfere with oocyte meiosis in a mouse model. Methods We divided mice into three groups: human recombinant centromere protein-A (human CENP-A, HA) and complete Freund’s adjuvant (CFA) were used to immunize mice for the study group (HA + CFA), and mice injected with CFA (CFA group) or saline (Saline group), respectively, served as controls. After immunization, serum anti-CENP-A antibody was detected by indirect immunofluorescence assay (IIFT) and enzyme-linked immunosorbent assay (ELISA). Chromosome alignment and intracellular IgG localization in MI- and MII-stage oocytes were investigated by immunofluorescence analysis. Results Positive ACAs were successfully induced by immunization with CENP-A and CFA, and results showed that the serum level of anti-CENP-A antibody was significantly higher in the HA + CFA group compared with the control groups. There was marked increase of chromosome misalignments in MI and MII oocytes in the HA + CFA group compared to the control groups. However, no oocytes from any of the three groups showed intracellular antibody immunofluorescence. Conclusions The development and maturation of oocytes were impaired in peripheral ACA positive mice, which exhibited severe chromosomal misalignments in metaphase meiosis; however, no evidence of ACAs entering the oocytes was observed, thus the underlying mechanism needs further exploration.

Systemic sclerosis: a rare presentation

Systemic sclerosis or so called scleroderma is an uncommon autoimmune inflammatory and fibrotic connective tissue disease involving multiple organs. The etiology of systemic sclerosis is currently unknown and its pathogenesis is only partially understood. Skin thickening and Raynaud’s phenomenon are the most common symptoms. Although systemic sclerosis is uncommon, it is associated with high morbidity and mortality. In this report, we present a case of a-43-year-old man with the complaint of weakness, tightening of the skin over the fingers, tingling in the soles of feet, nausea and significant weight loss. Laboratory examinations revealed positive ANA test, but negative anti topoisomerase I (anti-Scl-70), and anticentromere antibody.

Characteristics of clinical, laboratory, and immunological manifestations in patients with anticentromere antibody-associated Sjögren's disease

Objective: to study clinical and laboratory features in patients with anticentromere antibody (ACA)-positive SjЪgren's disease (SD), as well as the sensitivity of different methods for determination of ACA, and to elaborate an algorithm for differential diagnosis in ACA-positive patients.Patients and methods. The V.A. Nasonova Research Institute of Rheumatology followed up 136 patients who were highly positive for ACA. The investigators used the 2001 Russian criteria for the diagnosis for SD; the 2013 ACR/European League Against Rheumatism (EULAR) criteria for that of scleroderma systematica (SDS); the guidelines of the American Association for the Study of Liver Diseases, the Russian Gastroenterological Association, and the Russian Society for the Study of the Liver for that of primary biliary cholangitis (PBC)/biliary duct epitheliitis in the presence of SD. Lymphomas were diagnosed by biopsies of affected organs according to the WHO classification. SD was diagnosed in 119 patients; SDS in 49 cases (37 with SDS concurrent with SD and 12 with isolated SDS), PBC/biliary duct epitheliitis in 23 (all cases with PBC/biliary duct epitheliitis concurrent with SD and/or SDS); 5 patients were excluded from the investigation. Further analysis included 131 ACA-positive patients. The patients were divided into three groups: SD (n=82 or 62.6%); SD+SDS (n=37 or 28.24%); SDS (n=12 or 9.16%).Results and discussion. Autoantibodies to centromere peptide (CENP) A and CENP-B in the same titers were detected in all ACA-positive patients, regardless of diagnosis. Comparative analysis of three patient groups revealed no statistically significant differences in the frequency of laboratory deviations. The signs characteristic of classical SD (rheumatoid factor (RF)), anti-Ro and anti-La antibodies, leukopenia, higher ESR values, hypergammaglobulinemia, and elevated IgG/IgA levels) were found in a small proportion of patients. The frequency and severity of glandular manifestations did not differ in SD and SD + SDS. PBC/biliary duct epitheliitis was present in 17.5% of ACA-positive patients (in most antimitochondrial antibody-positive cases); no statistically significant differences in its frequency were found between the groups. Other extraglandular manifestations in SD and SD + SDS were identified in a smaller number of patients. All sclerodermic spectrum manifestations were more common in SD and SD + SDS than in BS. Pulmonary arterial hypertension was not diagnosed in any patient from the SD group. MALT lymphomas were detected in 19 ACA-positive patients. Those were present only in BS patients and absent in the SDS group. MALT lymphomas developed in the first 10 years after the onset of SD. The transformation of MALT lymphoma into diffuse large B-cell lymphoma was observed in 2 patients. The main signs of lymphomas in SD patients were persistent parotid salivary gland enlargement, decreased levels of complement C4 and peripheral blood CD19+ cells, as well as cryoglobulinemic vasculitis, serum monoclonal secretion, lymphoid infiltration in the minor salivary glands (a focus score of >4), and severe damage to the salivary and lacrimal glands.Conclusion. ACA-associated SD is an independent disease subtype characterized by an increased risk for SDS, PBC, and MALT lymphomas and by a low frequency of the systemic manifestations and laboratory signs characteristic of classical SD. Regardless of the detected type of antibodies and the presence or absence of extraglandular manifestations, damage to the salivary and lacrimal glands progresses in SD, which often leads to lymphomas; therefore, the therapy that may prevent this complication should be initiated as soon as possible after SD diagnosis. The lymphoproliferation signs identified in this investigation should be taken into account in all ACA-positive patients with SD for the early diagnosis of lymphoid tumors before therapy is prescribed. An algorithm for differential diagnosis in seropositivity for ACA is presented. Determination of autoantibodies to CENP-A and CENP-B does not allow the differential diagnosis in ACA-positive patients.

Comparison of Effect on Sicca Symptoms of Anticentromere Antibody–positive Sjögren Syndrome and Primary Sjögren Syndrome Alone

Objective.To determine whether positive anticentromere antibody (ACA) serology affects the severity of sicca symptoms in patients with primary Sjögren syndrome (pSS).Methods.Evaluation to detect subjective and objective sicca symptoms included questionnaires, physical examination, and pathology. Cases of pSS were classified according to the 2002 American-European Consensus Group (AECG) criteria. All patients were evaluated for presence of anti-Ro, anti-La, and ACA serology. Patients with pSS were categorized into ACA+ SS and ACA–SS. The groups were compared for measures of severity of oral and ocular sicca.Results.The pSS group had 446 patients, of whom 26 were ACA+ SS. Subjective ocular sicca measured 7.0 ± 2.4 (out of 10) in ACA+ SS and 6.4 ± 2.6 in ACA–SS (p = 0.197). Objective ocular sicca measured 3.2 mm ± 1.8 mm/5 min in ACA+ SS and 4.2 mm ± 4.4 mm/5 min in ACA–SS (p = 0.038). Subjective oral sicca measured 8.5 ± 1.4 in ACA+ SS and 6.7 ± 2.4 in ACA–SS (p < 0.001). Objective oral sicca measured 0.1 ml ± 0.2 ml/15 min in ACA+ SS and 0.4 ml ± 1.0 ml/15 min in ACA–SS (p < 0.001). Only 35% of ACA+ patients with SS were anti-Ro–positive or anti-La–positive compared with 77% of the ACA–patients with SS (p < 0.001). There was no significant difference in minor salivary gland fibrosis or focus scores between ACA+ SS and ACA–patients with SS.Conclusion.ACA+ SS is associated with more severe objective ocular sicca and more severe subjective and objective oral sicca compared to ACA–SS. The majority of ACA+ patients with SS meet AECG criteria for pSS despite negative serology for anti-Ro/La antibodies.

CLINICAL AND LABORATORY FEATURES OF ANTICENTROMERE ANTIBODY-POSITIVE SJö GREN’S SYNDROME

Objective: to study the clinical and laboratory features of patients with anticentromere antibody (ACA) positive Sjö gren’s syndrome (SjS); to assess the spectrum of autoantibodies in patients of this group; to determine the frequency with which the SjS patients who are highly positive for ACA, meet the international classification criteria for SjS and systemic sclerosis (SS); to reveal the incidence of MALT lymphomas in this patient group; to estimate the incidence of primary biliary cirrhosis (PBC)/biliary lesions as part of autoimmune epithelitis in SjS in this patient group.Material and methods. A total of 83 patients with ACA positive SjS were comprehensively examined at the V.A. Nasonova Research Institute of Rheumatology during the period 2012 to 2018. The inclusion criteria were con formity to the 2001 Russian SjS criteria and a high ACA level. MALT lymphomas were diagnosed on the basis of histological and immunohistochemical studies and polymerase chain reaction-based determination of B-cell clonality in the biopsy samples of affected organs according to the World Health Organization classification of Hematopoietic Tumors. The diagnosis of PBC/biliary lesions was made on the basis of histological and immunohistochemical studies of liver biopsy specimens.Results and discussion. The investigation revealed low detection rates for anti-Ro antibodies (32.5%), anti-La antibodies (7.2%) and rheumatoid factor (RF) (21.7%), which were typical for the classical SjS immunophenotype), increased ESR (14%), leukopenia (7%), hypergammaglobulinemia (17.6%), elevated levels of IgG (9.5%) and IgA (18.7%), and hypocomplementemia (16.1%) in the ACA positive SjS patients. Despite the low detection rate of RF, 15 (18%) patients in this group developed MALT lymphomas: 14 patients had salivary gland MALT lymphoma and one patient had tonsil MALT lymphoma with peripheral lymph node involvement (generalized marginal zone lymphoma). Also, the patients of this group showed high detection rates for AMA antibodies (34.6%), increased IgM level (29.7%) and a higher risk for PBC/biliary lesions as a manifestation of autoimmune epithelitis in SjS (14.5%). AMA-antibodies were absent in only two patients who were diagnosed with liver disease according to biopsy specimens. Nervous system and renal lesions, antiphospholipid syndrome, rheumatoid arthritis, hypergammaglobulinemic purpura, and cryoglobulinemic vasculitis were much less common and sporadic. Also ACA-positive SjS patients often have Raynaud’s phenomenon (54.9%) with scleroderma-type capillaroscopic changes (68%) and a limited form of SS (24%) according to the 2013 ACR criteria.Conclusion. ACA-positive SjS is a subtype of the disease, which is significantly different from the classic one in a number of clinical and laboratory signs and characterized by an increased risk for SS, MALT lymphomas, and PBC/biliary lesions as a manifestation of autoimmune epithelitis in SjS which in some cases leads to the underdiagnosis of SjS. ACA should be considered as pathogenetically related to SjS autoantibodies; and all patients who are seropositive for ACA should be examined for SjS and PBC/biliary lesions as a manifestation of autoimmune epithelitis in SjS regardless of whether they have SS or not, as well as complaints of dry mouth and eyes. Patients with significantly enlarged salivary glands should undergo biopsy to rule out or confirm MALT lymphoma before initiating hormonal, antilymphoproliferative, and anti-B-cell therapy.