Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Clinical Rheumatology
Published in: Clinical Rheumatology 5/2019

01-05-2019 | Original Article

Improved accuracy in DFS pattern interpretation using a novel HEp-2 ELITE system

Authors: Maria Infantino, O. Shovman, B. Gilburd, M. Manfredi, V. Grossi, Maurizio Benucci, A. Damiani, D. Chimenti, K. Malyavantham, Y. Shoenfeld

Published in: Clinical Rheumatology | Issue 5/2019

Login to get access

Abstract

Introduction/objectives

Accurate interpretation of DFS70 (dense fine speckled 70) and mixed antinuclear antibodies (ANAs) patterns can be challenging using conventional HEp-2 immunofluorescence (IIF) method. We evaluated a novel HEp-2 IIF substrate (HEp-2 ELITE/DFS70-KO) composed of a mixture of engineered HEp-2 devoid of the DFS70 autoantigen and conventional HEp-2 cells. The study assessed the utility of the new substrate in ANA screening and its advantages.

Method

One thousand and five consecutive routine samples sent for ANA screening were tested on both standard HEp-2 and the HEp-2 ELITE DFS70 KO substrates (ImmuGlo ANA HEp-2 and HEp-2 ELITE/DFS70-KO, Trinity Biotech, Buffalo, NY). Anti-DFS70 antibody specificity was additionally determined by immunoblot (IB). Clinical and serological data were included in the analysis of the overall impact of the novel HEp-2 substrate on DFS pattern interpretation.

Results

Of the 22 cases suspected as positive for DFS pattern alone or in combination with homogeneous or speckled patterns on conventional HEp-2 cells, 17 were interpreted with a higher accuracy using the new HEp-2 ELITE method as positive for DFS70 (monospecific DFS70 (10), mixed DFS70 (7)), speckled (3), and DFS (2) patterns.

Conclusions

The new substrate was not only useful in deciphering unclear mixed ANA patterns but also highly sensitive in detecting DFS70 pattern in comparison to the DFS70 positivity obtained using IB.
Literature
1.
2.
Chan EK, Damoiseaux J, Carballo OG, Conrad K, de Melo Cruvinel W, Francescantonio PL et al (2015) Report of the first international consensus on standardized nomenclature of antinuclear antibody HEp-2 cell patterns 2014-2015. Front Immunol 6:412CrossRefPubMedPubMedCentral
3.
Agmon-Levin N, Damoiseaux J, Kallenberg C, Sack U, Witte T, Herold M, Bossuyt X, Musset L, Cervera R, Plaza-Lopez A, Dias C, Sousa MJ, Radice A, Eriksson C, Hultgren O, Viander M, Khamashta M, Regenass S, Andrade LEC, Wiik A, Tincani A, Rönnelid J, Bloch DB, Fritzler MJ, Chan EKL, Garcia-de la Torre I, Konstantinov KN, Lahita R, Wilson M, Vainio O, Fabien N, Sinico RA, Meroni P, Shoenfeld Y (2014) International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies. Ann Rheum Dis 73:17–23CrossRefPubMed
4.
Meroni PL, Schur PH (2010) ANA screening: an old test with new recommendations. Ann Rheum Dis 69:1420–1422CrossRefPubMed
5.
Watanabe A, Kodera M, Sugiura K, Usuda T, Tan EM, Takasaki Y, Tomita Y, Muro Y (2004) Anti-DFS70antibodies in 597 healthy hospital workers. Arthritis Rheum 50:892–900CrossRefPubMed
6.
Malyavantham KS, Suresh L (2018) Simultaneous distinction of Monospecific and mixed DFS70 patterns during ANA screening with a novel HEp-2 ELITE/DFS70 knockout substrate. J Vis Exp 131
7.
Malyavantham KL, Suresh L (2017) Analysis of DFS70 pattern and impact on ANA screening using a novel HEp-2 ELITE/DFS70 knockout substrate. Auto Immun Highlights 8:3CrossRefPubMedPubMedCentral
8.
9.
Basu A, Sanchez TW, Casiano CA (2015) DFS70/LEDGF p75: an enigmatic autoantigen at the interface between autoimmunity, AIDS, andcancer. Front Immunol 6:116CrossRefPubMedPubMedCentral
10.
Ochs RL, Muro Y, Si Y, Ge H, Chan EK, Tan EM (2000) Autoantibodies to DFS 70 kd/transcription coactivator p75 in atopic dermatitis and other conditions. J Allergy Clin Immunol 105:1211–1220CrossRefPubMed
11.
Infantino M, Meacci F, Grossi V, Manfredi M, Li Gobbi F, Sarzi-Puttini P, Atzeni F, Benucci M (2017) The clinical impact of anti-DFS70 antibodies in undifferentiated connective tissue disease: case reports and a review of the literature. Immunol Res 65:293–295CrossRefPubMed
12.
Seelig CA, Bauer O, Seelig HP (2016) Autoantibodies against DFS70/LEDGF exclusion markers for systemic autoimmune rheumatic diseases (SARD). Clin Lab 62:499–517CrossRefPubMed
13.
Mariz HA, Sato EI, Barbosa SH, Rodrigues SH, Dellavance A, Andrade LE (2011) Pattern on the antinuclear antibody-HEp-2 test is a critical parameter for discriminating antinuclear antibody-positive healthy individuals and patients with autoimmune rheumatic diseases. Arthritis Rheum 63:191–200CrossRefPubMed
14.
Mahler M, Parker T, Peebles CL Andrade LE, Swart A, Carbone Y et al (2012) Anti-DFS70/LEDGF antibodies are more prevalent in healthy individuals compared to patients with systemic autoimmune rheumatic diseases. J Rheumatol 39:2104–2110CrossRefPubMed
15.
Bizzaro N, Tonutti E, Tampoia M, Infantino M, Cucchiaro F, Pesente F, Morozzi G, Fabris M, Villalta D (2015) Specific chemoluminescence and immunoadsorption tests for anti-DFS70 antibodies avoid false positive results by indirect immunofluorescence. Clin Chim Acta 451:271–277CrossRefPubMed
16.
Nilsson AC, Voss A, Lillevang ST (2015) DFS70 autoantibodies are rare in healthy Danish individuals but may still serve as a diagnostic aid. Scand J Immunol 82:547–548CrossRefPubMed
17.
Conrad K, Rober N, Andrade LE, Mahler M (2017) The clinical relevance ofanti-DFS70 autoantibodies. Clin Rev Allergy Immunol 52:202–216CrossRefPubMed
18.
Fritzler MJ (2016) Choosing wisely: review and commentary on anti-nuclear antibody (ANA) testing. Autoimmun Rev 15:272–280CrossRefPubMed
19.
Gundín S, Irure-Ventura J, Asensio E, Ramos D, Mahler M, Martínez-Taboada V, López-Hoyos M (2016) Measurement of anti-DFS70 antibodies in patients with ANA-associated autoimmune rheumatic diseases suspicion is cost-effective. Auto Immun Highlights 7:10CrossRefPubMedPubMedCentral
20.
Carter JB, Carter S, Saschenbrecker S, Goeckeritz BE (2018) Recognition and relevance of anti-DFS70 autoantibodies in routine antinuclear autoantibodies testing at a community hospital. Front Med (Lausanne) 5:88CrossRef
21.
Shovman O, Gilburd B, Chayat C, Amital H, Langevitz P, Watad A, Guy A, Perez D, Azoulay D, Blank M, Segal Y, Bentow C, Mahler M, Shoenfeld Y (2018) Prevalence of anti-DFS70 antibodies in patients with and without systemic autoimmune rheumatic diseases. Clin Exp Rheumatol 36:121–126PubMed
22.
23.
Ogawa Y, Sugiura K, Watanabe A, Kunimatsu M, Mishima M, Tomita Y, Muro Y (2004) Autoantigenicity of DFS70 is restricted to the conformational epitope of C-terminal alpha-helical domain. J Autoimmun 23:221–231CrossRefPubMed
24.
Muro Y, Sugiura K, Morita Y, Tomita Y (2008) High concomitance of disease marker autoantibodies in anti-DFS70/LEDGF autoantibody-positive patients with autoimmune rheumatic disease. Lupus 17:171–176CrossRefPubMed
25.
Bentow C, Fritzler MJ, Mummert E, Mahler M et al (2016) Recognition of the dense fine speckled (DFS) pattern remains challenging: results from an international internet-based survey. Auto Immun Highlights 7:8CrossRefPubMedPubMedCentral
26.
Bentow C, Rosenblum R, Correia P, Karayev E, Karayev D, Williams D, Kulczycka J, Fritzler MJ, Mahler M (2016) Development and multi-center evaluation of a novel immunoadsorption method for anti-DFS70 antibodies. Lupus 25:897–904CrossRefPubMed
27.
Mutlu E, Eyigör M, Mutlu D, Gültekin M (2016) Confirmation of anti-DFS70 antibodies is needed in routine clinical samples with DFS staining pattern. Cent Eur J Immunol 41:6–11CrossRefPubMedPubMedCentral
28.
Rigon A, Buzzulini F, Soda P, Onofri L, Arcarese L, Iannello G, Afeltra A (2011) Novel opportunities in automated classification of antinuclear antibodies on HEp-2 cells. Autoimmun Rev 10:647–652CrossRefPubMed
29.
Bizzaro N, Tonutti E, Villalta D (2011) Recognizing the dense fine speckled/lens epithelium-derived growth factor/p75 pattern on HEP-2 cells: not an easy task! Comment on the article by Mariz et al. Arthritis Rheum 63:4036–4037 author reply 4037–8CrossRefPubMed
30.
Mahler M, Meroni PL, Andrade LE, Khamashta M, Bizzaro N, Casiano CA, Fritzler MJ (2016) Towards a better understanding of the clinical association of anti-DFS70 autoantibodies. Autoimmun Rev 15:198–201CrossRefPubMed
31.
Bizzaro N, Pesente F, Cucchiaro F, Infantino M, Tampoia M, Villalta D, Fabris M, Tonutti E (2016) Anti-DFS70 antibodies detected by immunoblot methods: a reliable tool to confirm the dense fine speckles ANA pattern. J Immunol Methods 436:50–53CrossRefPubMed
32.
Shovman O, Gilburd B, Barzilai O, Shinar E, Larida B, Zandman-Goddard G et al (2005) Evaluation of the BioPlex 2200 ANA screen: analysis of 510 healthy subjects: incidence of natural/predictive autoantibodies. Ann N Y Acad Sci 1050:380–388CrossRefPubMed
Metadata
Title
Improved accuracy in DFS pattern interpretation using a novel HEp-2 ELITE system
Authors
Maria Infantino
O. Shovman
B. Gilburd
M. Manfredi
V. Grossi
Maurizio Benucci
A. Damiani
D. Chimenti
K. Malyavantham
Y. Shoenfeld
Publication date
01-05-2019
Publisher
Springer London
Published in
Clinical Rheumatology / Issue 5/2019
Print ISSN: 0770-3198
Electronic ISSN: 1434-9949
DOI
https://doi.org/10.1007/s10067-018-04412-1

Other articles of this Issue 5/2019

Clinical Rheumatology 5/2019 Go to the issue

Brief Report

Venous vessel wall thickness in lower extremity is increased in male patients with Behcet’s disease

Editorial

Biosimilars in the Americas: the future by consensus

Original Article

Functional implications of single nucleotide polymorphisms rs662 and rs854860 on the antioxidative activity of paraoxonase1 (PON1) in patients with rheumatoid arthritis

Original Article

First trimester combined screening in patients with systemic lupus erythematosus: impact of pre-analytical variables on risk assessment

Review Article

Accuracy of neutrophil CD64 expression in diagnosing infection in patients with autoimmune diseases: a meta-analysis

Original Article

Usefulness of rheumatoid factor as an immunological and prognostic marker in PSS patients
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits