Store all reagents at 2-8°C. All reagents must be brought to room temperature (20-25°C) prior to use. When stored at 2-8°C, the diluted Assay wash buffer is stable until the kit expiration date.
General Description
Antibodies to the SSA antigen (also known as Ro antigen) are one of the most frequent serological markers of autoimmunity in rheumatic diseases. They appear in 60-70% of patients with Sjögren's syndrome (SS), 30-40% of patients with systemic lupus erythematosus (SLE), and 3-5% of patients with rheumatoid arthritis (RA). The SSA antigen is comprised of an acidic 60 kDa protein that may exist in complex with RNA (80-112 bases). However, RNA is not required for SSA antigenicity. The SSA antigen is predominantly cytoplasmic, and not located in the nucleus. Therefore, patients with antibody to SSA may be ANA negative on routine testing. Approximately 62% of ANA negative lupus patients have antibodies to SSA. Two types of anti-Ro/SSA antibodies have been identified; Ro/SSA antigens of 60 kDa and 52 kDa. Anti-SSA-60 kDa antibodies are linked to certain disorders such as SS, SLE, neonatal lupus and congenital heart block. Clinically, the presence of aSSA52 has been reported in a wide variety of diseases, includes inflammatory myositis, primary biliary cirrhosis and SS.
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Background
The Ro/La system is considered to be a heterogeneous antigen complex, which consists of three different proteins (52kDa Ro, 60kDa Ro and La) and four small RNA particles. Anti-Ro/SSA antibodies are the most common specific antibodies in many autoimmune diseases such as systemic lupus erythematosus (SLE), SS/SLE overlap syndrome, subacute cutaneous lupus erythematosus (SCLE), neonatal lupus erythematosus and primary biliary cirrhosis. The differences between 52kDa, 60kDa Ro and La may explain why different detection methods differ in the detection of anti-Ro and anti-La autoantibodies.
Anti-Ro/SSA-antibodies, including anti-Ro/SSA-52kD and anti-Ro/SSA-60kD subspecificity, are autoimmune responses to two subunits of intracellular ribonucleoprotein Ro (52kD and 60kD Ro). They are polyclonal antibodies, usually belonging to the IgG class, and are common in patients with autoimmune diseases (AD) and other patients. In these diseases, anti-Ro/SSA-60kD subspecificity has a more definite direct pathogenic role than anti-Ro-52kD subspecificity in the occurrence of classical autoimmune manifestations, and is also related to a higher incidence of extraglandular features (especially vasculitis) and stronger systemic activity. A large number of studies have shown that anti-Ro/SSA antibodies can also be detected in a considerable portion of the general population (0.5%-2.7%), and they have no AD symptoms in most cases (60%). In addition, the entry of anti-Ro/SSA antibodies from the mother to the fetus through the placenta can lead to autoimmune congenital heart block (aCHB), which is a typical passive acquired autoimmunity. Although the mechanism of this disease is not clear, many clinical and experimental data have proved that the inhibitory cross-reaction between anti-Ro/SSA antibodies and L-type and T-type calcium channels in fetal conduction system cardiomyocytes plays a key role.
Figure 1. The anti-Ro/SSA 52 antibodies interact with calcium channels in both adults and fetuses (Source: Santos-Pardo I, et al. 2015)
Our product [DEIA103J] detects anti-SSA antibody (Ro60) in human serum, which is based on the principle of solid-phase enzyme-linked immunosorbent assay using SSA protein immobilized on a microtiter plate using a secondary antibody labeled with HRP.
References
1. Lazzerini PE, et al. Anti-Ro/SSA Antibodies and the Autoimmune Long-QT Syndrome. Front Med (Lausanne). 2021 Sep 6;8:730161.
2. Lazzerini PE, et al. Anti-Ro/SSA Antibodies Blocking Calcium Channels as a Potentially Reversible Cause of Atrioventricular Block in Adults. JACC Clin Electrophysiol. 2023 Aug;9(8 Pt 3):1631-1648.
3. Santos-Pardo I, et al. Anti-Ro/SSA antibodies and cardiac rhythm disturbances: Present and future perspectives. Int J Cardiol. 2015 Apr 1;184:244-250.
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References
Anti-Ro/SSA Antibodies and the Autoimmune Long-QT Syndrome
Front Med (Lausanne)
Authors: Lazzerini PE, Laghi-Pasini F, Boutjdir M, Capecchi PL.
Autoimmunity is increasingly recognized as a novel pathogenic mechanism for cardiac arrhythmias. Several arrhythmogenic autoantibodies have been identified, cross-reacting with different types of surface proteins critically involved in the cardiomyocyte electrophysiology, primarily ion channels (autoimmune cardiac channelopathies). Specifically, some of these autoantibodies can prolong the action potential duration leading to acquired long-QT syndrome (LQTS), a condition known to increase the risk of life-threatening ventricular arrhythmias, particularly Torsades de Pointes (TdP). The most investigated form of autoimmune LQTS is associated with the presence of circulating anti-Ro/SSA-antibodies, frequently found in patients with autoimmune diseases (AD), but also in a significant proportion of apparently healthy subjects of the general population. Accumulating evidence indicates that anti-Ro/SSA-antibodies can markedly delay the ventricular repolarization via a direct inhibitory cross-reaction with the extracellular pore region of the human-ether-a-go-go-related (hERG) potassium channel, resulting in a higher propensity for anti-Ro/SSA-positive subjects to develop LQTS and ventricular arrhythmias/TdP. Recent population data demonstrate that the risk of LQTS in subjects with circulating anti-Ro/SSA antibodies is significantly increased independent of a history of overt AD, intriguingly suggesting that these autoantibodies may silently contribute to a number of cases of ventricular arrhythmias and cardiac arrest in the general population. In this review, we highlight the current knowledge in this topic providing complementary basic, clinical and population health perspectives.
Anti-Ro/SSA and/or anti-La/SSB antibodies are associated with adverse IVF and pregnancy outcomes
J Reprod Immunol
Authors: Fang X, Lu F, Wang Y, Guo L, Zhang Y, Bai S, Kwak-Kim J, Wu L.
Anti-Ro/SSA and/or anti-La/SSB antibodies (anti-SSA/SSB) were reported to increase the risk of recurrent pregnancy loss. However, the effects of anti-SSA/SSB antibodies on in-vitro fertilization (IVF) and pregnancy outcomes were still unclear. The purpose of the study was to determine whether anti-SSA/SSB antibodies were detrimental to IVF and pregnancy outcomes. This study included 55 anti-SSA/SSB antibodies-positive women and 61 anti-SSA/SSB antibodies-negative control women receiving gonadotropin-releasing hormone (GnRH) agonist long protocol (n = 30 and 39, respectively) or GnRH antagonist protocol (n = 25 and 22, respectively) for in-vitro fertilization and embryo transfer (IVF-ET). The impact of anti-SSA/SSB antibodies on immune-related indicators, fertilization, embryo development and pregnancy outcomes were analyzed. With either GnRH agonist or antagonist protocol, women with anti-SSA/SSB had higher levels of peripheral blood cytokines, including TNF-α and IL-17A, lower levels of peripheral blood Th and NK cells, and poor IVF outcomes, including lower number of retrieved oocytes and embryos, lower M II oocytes rate, blastocyst formation rate, and perfect and available embryo rates. Moreover, clinical pregnancy rate, implantation rate, take-home baby rate, and birth weight were significantly lower in the study group as compared with those of the control group. In conclusion, women with anti-SSA/SSB are associated with adverse IVF and pregnancy outcomes. Screening for these antibodies and proper counselling of couples undergoing IVF-ET should be considered. Underlying immunopathology associated with SSA/SSB antibodies and reproduction should be explored further.
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Figure 1. The anti-Ro/SSA 52 antibodies interact with calcium channels in both adults and fetuses
