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
- •
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.
- •
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.
- •
Adsorption of anti-DFS70
References (27)
- et al.
Recent advances in diagnostic technologies for autoimmune diseases
Autoimmun Rev
(2009) - et al.
Autoantibodies to DFS 70 kd/transcription coactivator p75 in atopic dermatitis and other conditions
J Allergy Clin Immunol
(2000) - et al.
Humoral immune response directed against LEDGF in patients with VKH
Immunol Lett
(2001) Challenges to the use of autoantibodies as predictors of disease onset, diagnosis and outcomes
Autoimmun Rev
(2008)- et al.
Predictive value of antinuclear autoantibodies: the lessons of the systemic sclerosis autoantibodies
Autoimmun Rev
(2008) - et al.
HLA-associated production of anti-DFS70/LEDGF autoantibodies and systemic autoimmune disease
J Autoimmun
(2006) - et al.
Autoantibodies in interstitial cystitis
J Urol
(1994) - et al.
LEDGF/p75 is essential for nuclear and chromosomal targeting of HIV-1 integrase in human cells
J Biol Chem
(2003) - et al.
Challenges of automated screening and differentiation of non-organ specific autoantibodies on HEp-2 cells
Autoimmun Rev
(2009) - et al.
Novel opportunities in automated classification of antinuclear antibodies on HEp-2 cells
Autoimmun Rev
(2011)
Italian multicentre study for application of a diagnostic algorithm in autoantibody testing for autoimmune rheumatic disease: conclusive results
Autoimmun Rev
Epitope specificity and significance in systemic autoimmune diseases
Ann N Y Acad Sci
Detection of antinuclear antibodies by indirect immunofluorescence and by solid phase assay
Autoimmun Rev
Cited by (95)
Benign prostatic hyperplasia - A novel autoimmune disease with a potential therapy consequence?
2024, Autoimmunity ReviewsInvestigation of dense fine speckled pattern and anti-dense fine speckled 70 antibody by a single step assay
2022, Journal of Microbiological MethodsAutoantibodies to dense-fine-speckled 70 (DFS70) do not necessarily rule out connective tissue diseases
2022, Seminars in Arthritis and RheumatismThe 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