ReviewImportance of the dense fine speckled pattern on HEp-2 cells and anti-DFS70 antibodies for the diagnosis of systemic autoimmune diseases
Section snippets
History and clinical association of anti-DFS70 antibodies
The presence of autoantibodies directed against intracellular antigens, including anti-nuclear antibodies (ANA), is a hallmark of systemic autoimmune rheumatic diseases (SARD) [1]. Despite significant advances in diagnostic technologies [2], [3], the indirect immunofluorescence (IIF) assay using HEp-2 cells continues to be one of the most commonly used routine tests for the detection of ANA. This is reflected in the recommendations of a task force commissioned by the American College of
Indirect immunofluorescence picture of anti-DFS70 antibodies and cellular function of LEDGF
The typical IIF staining pattern has been described as dense fine speckles that are somewhat uniformly distributed throughout the nucleus accompanied by metaphase chromatin staining [19] (Fig. 1). Since a 70-kDa protein was recognized by immunoblotting, the antigen was initially termed DFS70, but the primary target autoantigen was later identified as the lens epithelium-derived growth factor (LEDGF) [20], now known to be DNA binding transcription coactivator p75 (reviewed in Ref. [9]). This
Advances in ANA testing by indirect immunofluorescence
Several improvements have been made to the ANA HEp-2 assay including the use of cell lines that over-express target antigens after transfection of a cognate cDNA [22], antigen supplements in the form of micro dots, and optimized mitotic cell lines. At the technical platform level, digital imaging systems for automated reading, archiving and interpretation of the results have been reported to provide good positive/negative discrimination [23], [24]. In addition, promising results have been
Consequences for ANA testing — a new algorithm
With advances in autoimmunity research and the availability of new autoantibody assays, it is important to develop and implement novel test algorithms for the diagnosis of SARD. A close collaboration between clinicians and laboratory specialists is required to increase the efficiency, performance and validity of any testing cascade. Recently, using a multi-center approach, a novel consensus test algorithm has been established and implemented in a broad geographic area in Italy [25]. However,
Immunoadsorption of anti-DFS70 antibodies
In a recent study, the DFS IIF pattern was found in 33.1% of ANA positive healthy individuals compared to 0.0% of ANA positive patients with SARD (p < 0.0001), which significantly affects the diagnostic power and efficiency of the IIF assay. Therefore, accurate pattern recognition and appropriate interpretation and reporting of results are important because it could influence the referral of patients with a positive ANA for unnecessary tertiary care consultation. Since the identification of the
Abbreviations
- ANA
anti-nuclear antibody
- DFS
dense fine speckled
- HI
healthy individuals
- IIF
indirect immunofluorescence
- SARD
systemic autoimmune rheumatic disease
- SLE
systemic lupus erythematosus
Disclosure statement
M. Mahler is employed at INOVA Diagnostics Inc. selling antibody assays to aid in the diagnosis of autoimmune diseases. M.J. Fritzler is a consultant to ImmunoConcepts, BioRad and INOVA Diagnostics. J.G. Hanly has no conflict of interest.
Take-home messages
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The dense fine speckled (DFS) pattern is rarely found in isolation in systemic autoimmune rheumatic disease sera, but frequently in sera from apparently healthy individuals. Therefore, it is important to recognize this pattern and confirm the reactivity to DFS70 by a specific assay.
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The DFS pattern on ANA IIF and the testing for anti-DFS70 antibodies need to be considered in diagnostic algorithms of ANA testing in the context of SARD — a new algorithm has been proposed.
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Adsorption of anti-DFS70
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2024, Autoimmunity ReviewsInvestigation of dense fine speckled pattern and anti-dense fine speckled 70 antibody by a single step assay
2022, Journal of Microbiological MethodsCitation Excerpt :Specific commercial tests like ELISA, chemiluminescent assay (CLIA), line-blot, and dot-blot tests to confirm the presence of anti-DFS70 antibodies are available. In addition to these methods, two modified IIF methods which were based on immunoadsorption and evaluation on DFS70 knock-out substrate were developed (Mahler et al., 2012b; Bentow et al., 2016b). In 2012, Mahler et al. developed a novel immunoadsorption method and observed the diminishing of the DFS staining pattern on HEp-2 substrate after adsorption of the anti-DFS70 positive sera with the recombinant DFS70 antigen fragment.
Improved performance of confirmatory assays for detecting dense fine speckled (DFS) 70 antibodies
2022, PathologyCitation Excerpt :Studies also found that the prevalence of these antibodies in patients with SARD was very low but significantly higher in healthy individuals.12–16 Increasing reports have suggested that monospecific DFS70 autoantibodies without concomitant SARD-associated antibodies could be an exclusion biomarker for SARD diagnosis.12–14,16–18 The typical DFS pattern depicts dense fine speckled fluorescence staining distributed throughout interphase nucleus and metaphase chromatin, but negative in nucleoli.
Autoantibodies to dense-fine-speckled 70 (DFS70) do not necessarily rule out connective tissue diseases
2022, Seminars in Arthritis and RheumatismCitation Excerpt :However, the presence of ANA can also be detected in up to 20% of healthy individuals [3]. One reason is the presence of anti-dense-fine-speckled-70 antibodies [3]. The term DFS70 evolved from the specific dense fine speckled pattern in IIF and its antigen with a molecular weight of 70 kDa in line immunoassay [4].
The antinuclear antibody dense fine speckled pattern and possible clinical associations: An indication of a proinflammatory microenvironment
2021, Journal of Immunological MethodsClinical value of anti-DFS70 antibodies in a cohort of patients undergoing routine antinuclear antibodies testing
2020, Journal of Immunological Methods