Abstract
BackgroundDetection of autoantibodies giving nuclear rim pattern by immunofluorescence (anti-nuclear envelope antibodies - ANEA) in sera from patients with primary biliary cirrhosis (PBC) is a useful tool for the diagnosis and prognosis of the disease. Differences in the prevalence of ANEA in PBC sera so far reported have been attributed to the methodology used for the detection as well as to ethnic/geographical variations. Therefore, we evaluated the prevalence of ANEA in sera of Greek patients with PBC by using methods widely used by clinical laboratories and a combination of techniques and materials.MethodsWe screened 103 sera by immunoblotting on nuclear envelopes and indirect immunofluorescence (IIF) using cells and purified nuclei. Reactivities against specific autoantigens were assessed using purified proteins, ELISA, immunoprecipitation and mass spectrometry.ResultsWe found higher prevalence of ANEA when sera were assayed by IIF on purified nuclei or cultured cells (50%) compared to Hep2 commercially available slides (15%). Anti-gp210 antibodies were identified in 22.3% and 33% of sera using ELISA for the C-terminal of gp210 or both ELISA and immunoprecipitation, respectively. Immunoblotting on nuclear envelopes revealed that immunoreactivity for the 210 kDa zone is related to anti-gp210 antibodies (p < 0.0001). Moreover, we found that sera had antibodies for lamins A (6.8%), B (1%) and C (1%) and LBR (8.7%), whereas none at all had detectable anti-p62 antibodies.ConclusionsThe prevalence of ANEA or anti-gp210 antibodies is under-estimated in PBC sera which are analyzed by conventional commercially available IIF or ELISA, respectively. Therefore, new substrates for IIF and ELISA should be included by clinical laboratories in the analysis of ANEA in autoimmune sera.
Highlights
Detection of autoantibodies giving nuclear rim pattern by immunofluorescence in sera from patients with primary biliary cirrhosis (PBC) is a useful tool for the diagnosis and prognosis of the disease
We report that against nuclear envelope (ANEA) and anti-gp210 are present with high frequency in PBC sera
In PBC sera, it has been shown that the prevalence of antibodies giving a rim-like membranous fluorescence (10%) by indirect immunofluorescence (IIF) on Hep2 cells was lower than the prevalence of antibodies assayed by ELISA against gp210 (16%), which represent one among other autoantigens of the nuclear envelope
Summary
Detection of autoantibodies giving nuclear rim pattern by immunofluorescence (anti-nuclear envelope antibodies - ANEA) in sera from patients with primary biliary cirrhosis (PBC) is a useful tool for the diagnosis and prognosis of the disease. The nuclear lamina is composed of A- and B-type lamins These proteins form a polymeric lining that supports the inner nuclear membrane and imparts elasticity to. A significant variation of their prevalence (between 10% and 48%) has been reported, when indirect immunofluorescence (IIF) is used for the screening of PBC sera [4,6,7,8,9,10,11] This could be attributed to differences on the processing of IIF samples, namely substrates and reagents used for the detection and on the evaluation of the results, especially when antibodies of cytoplasmic specificities are present in the same serum [3,4,11]
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