Skip to main content

Advertisement

SpringerLink
Log in

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

  • Original Article
  • Published:
Clinical Rheumatology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. American College of Rheumatology Position Statement (2011) Methodology of testing for antinuclear antibodies. www.rheumatology.org/practice/ana_position_stmt.pdf. Approved by Board of Directors: Aug 2011

  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 6:412

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 73:17–23

    Article  CAS  PubMed  Google Scholar 

  4. Meroni PL, Schur PH (2010) ANA screening: an old test with new recommendations. Ann Rheum Dis 69:1420–1422

    Article  CAS  PubMed  Google Scholar 

  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 50:892–900

    Article  CAS  PubMed  Google Scholar 

  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 8:3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Fritzler MJ (2011) The antinuclear antibody test: last or lasting gasp? Arthritis Rheum 63:19–22

    Article  PubMed  Google Scholar 

  9. Basu A, Sanchez TW, Casiano CA (2015) DFS70/LEDGF p75: an enigmatic autoantigen at the interface between autoimmunity, AIDS, andcancer. Front Immunol 6:116

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 105:1211–1220

    Article  CAS  PubMed  Google Scholar 

  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 65:293–295

    Article  CAS  PubMed  Google Scholar 

  12. Seelig CA, Bauer O, Seelig HP (2016) Autoantibodies against DFS70/LEDGF exclusion markers for systemic autoimmune rheumatic diseases (SARD). Clin Lab 62:499–517

    Article  CAS  PubMed  Google Scholar 

  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 63:191–200

    Article  CAS  PubMed  Google Scholar 

  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 39:2104–2110

    Article  PubMed  Google Scholar 

  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 451:271–277

    Article  CAS  PubMed  Google Scholar 

  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 82:547–548

    Article  CAS  PubMed  Google Scholar 

  17. Conrad K, Rober N, Andrade LE, Mahler M (2017) The clinical relevance ofanti-DFS70 autoantibodies. Clin Rev Allergy Immunol 52:202–216

    Article  CAS  PubMed  Google Scholar 

  18. Fritzler MJ (2016) Choosing wisely: review and commentary on anti-nuclear antibody (ANA) testing. Autoimmun Rev 15:272–280

    Article  PubMed  Google Scholar 

  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 7:10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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) 5:88

    Article  Google Scholar 

  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 36:121–126

    PubMed  Google Scholar 

  22. Infantino M, Shovman O, Pérez D, Manfredi M, Grossi V, Benucci M et al (2018) Anti-DFS70 autoantibodies in undifferentiated connective tissue diseases subjects: what’s on the horizon? Rheumatology (Oxford). https://doi.org/10.1093/rheumatology/key012

  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 23:221–231

    Article  CAS  PubMed  Google Scholar 

  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 17:171–176

    Article  CAS  PubMed  Google Scholar 

  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 7:8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 25:897–904

    Article  CAS  PubMed  Google Scholar 

  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 41:6–11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 10:647–652

    Article  CAS  PubMed  Google Scholar 

  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 4037–8

    Article  PubMed  Google Scholar 

  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 15:198–201

    Article  CAS  PubMed  Google Scholar 

  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 436:50–53

    Article  CAS  PubMed  Google Scholar 

  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 1050:380–388

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Immunology and Allergology Laboratory Unit, S. Giovanni di Dio Hospital, Azienda Usl Toscana Centro, Via Torregalli, 3, 50143, Florence, Italy

    Maria Infantino, M. Manfredi & V. Grossi

  2. Zabludowitz Center for Autoimmune Diseases, Sheba Medical Center, Ramat Gan, Israel

    O. Shovman, B. Gilburd & Y. Shoenfeld

  3. Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

    O. Shovman & Y. Shoenfeld

  4. Rheumatology Unit, S. Giovanni di Dio Hospital, Azienda Usl Toscana Centro, Florence, Italy

    Maurizio Benucci & A. Damiani

  5. A.Menarini Diagnostics, Florence, Italy

    D. Chimenti

  6. Trinity Biotech, Buffalo, NY, USA

    K. Malyavantham

  7. Incumbent of the Laura Schwarz-Kipp Chair for Research of Autoimmune Diseases, Tel Aviv University, Tel Aviv, Israel

    Y. Shoenfeld

Authors
  1. Maria Infantino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. O. Shovman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B. Gilburd
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Manfredi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. V. Grossi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maurizio Benucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. Damiani
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. D. Chimenti
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K. Malyavantham
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Y. Shoenfeld
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maria Infantino.

Ethics declarations

Disclosures

None.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Infantino, M., Shovman, O., Gilburd, B. et al. Improved accuracy in DFS pattern interpretation using a novel HEp-2 ELITE system. Clin Rheumatol 38, 1293–1299 (2019). https://doi.org/10.1007/s10067-018-04412-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10067-018-04412-1

Keywords

Search

Navigation