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Changes in the result of antinuclear antibody immunofluorescence assay on HEp-2 cells reflect disease activity status in systemic lupus erythematosus

  • Mônica Simon Prado , Alessandra Dellavance , Silvia Helena Rodrigues , Valdecir Marvulle and Luis Eduardo Coelho Andrade ORCID logo EMAIL logo
Published/Copyright: March 12, 2020
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 8

Abstract

Background

The objective of the study was to determine whether the staining pattern and titer of indirect immunofluorescence assay on HEp-2 cells (HEp-2 IFA) are associated with systemic lupus erythematosus (SLE) disease activity.

Methods

A total of 269 consecutive patients meeting the ACR and SLICC criteria for SLE were classified into three groups according to the SLE Disease Activity Index 2000 (SLEDAI2K): Remission (SLEDAI2K = 0; n = 47); Intermediate (SLEDAI2K = 1-5; n = 111); Active (SLEDAI2K ≥ 6; n = 111). All subjects were assessed for HEp-2 IFA titer and staining pattern and nine traditional parameters of SLE disease activity. After a 6 to 12-month interval, 101 of the 269 patients were reassessed.

Results

HEp-2 IFA homogeneous nuclear pattern (AC-1) occurred more frequently in the Active Group compared to the Remission Group (p < 0.001). Fine speckled nuclear pattern (AC-4) tended to occur more frequently in the Remission Group compared to the Active Group (p = 0.054). Subjects with AC-1 pattern had higher SLEDAI (8.8 ± 7.6) than those with AC-4 (4.8 ± 5.2) (p < 0.001). HEp-2 IFA titer and anti-nuclear antibody by enzyme-linked immunosorbent assay (ANA-ELISA) values were lower in the Remission Group compared to the other two groups (p < 0.001). Multivariate analyses identified only ELISA anti-dsDNA as an independent variable associated with disease activity. In follow-up analysis, HEp-2 IFA titer decreased significantly in the 33 subjects with decreased disease activity (p = 0.002). Receiver operator characteristic (ROC) curve analysis for determination of disease activity showed equivalent areas under the curve (AUC) for HEp-2 IFA titer and traditional disease activity parameters.

Conclusions

HEp-2 IFA pattern and titer can reflect SLE disease activity and may be considered in conjunction with other laboratory and clinical parameters in the assessment of SLE disease activity.

Keywords: antinuclear antibody; autoantibodies; disease activity; HEp-2 cells; SLEDAI2K; systemic lupus erythematosus
Corresponding author: Luis Eduardo Coelho Andrade, MD, PhD, Rheumatology Division, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Botucatu 740, 3o andar, São Paulo, SP 04023-062, Brazil; and Immunology Division, Fleury Medicine and Health Laboratories, Sao Paulo, Brazil, Phone/Fax: +55 11 5576-4239

Funding source: CNPq

Award Identifier / Grant number: 232711/2014-3

Funding source: FAPESP

Award Identifier / Grant number: 2013/00913-6

Funding statement: This work was supported by the Brazilian government research foundation CNPq (National Council for Research and Technology) [grant 232711/2014-3, Funder Id: http://dx.doi.org/10.13039/501100003593] and by the Sao Paulo Government agency FAPESP (Sao Paulo State Research Foundation) [grant 2013/00913-6, Funder Id: http://dx.doi.org/10.13039/501100001807].

Acknowledgments

The authors thank Mary Chaiken, PhD, for her expert review of the English.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Employment or leadership: None declared.

  3. Honorarium: None declared.

  4. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. La Paglia GM, Leone MC, Lepri G, Vagelli R, Valentini E, Alunno A, et al. One year in review 2017: systemic lupus erythematosus. Clin Exp Rheumatol 2017;35:551–61.Search in Google Scholar

2. Dvorkina O, Ginzler EM. Clinical features of systemic lupus erythematosus. In: Hochberg MC, Silman AJ, Smolen JS, Weinblat ME, Weisman MH, editors. Rheumatology. Philadelphia, PA: Mosby Elsevier, 2015:1032–44.10.1016/B978-0-323-09138-1.00126-1Search in Google Scholar

3. Griffiths B, Mosca M, Gordon C. Assessment of patients with systemic lupus erythematosus and the use of lupus disease activity indices. Best Pract Res Clin Rheumatol 2005;19:685–708.10.1016/j.berh.2005.03.010Search in Google Scholar PubMed

4. Ruperto N, Hanrahan LM, Alarcon GS, Belmont HM, Brey RL, Brunetta P, et al. International consensus for a definition of disease flare in lupus. Lupus 2011;20:453–62.10.1177/0961203310388445Search in Google Scholar PubMed

5. Nuttall A, Isenberg DA. Assessment of disease activity, damage and quality of life in systemic lupus erythematosus: new aspects. Best Pract Res Clin Rheumatol 2013;27:309–18.10.1016/j.berh.2013.10.003Search in Google Scholar PubMed

6. Jung JY, Bae CB, Suh CH. Promising biomarkers for systemic lupus erythematosus. Expert Opin Med Diagn 2013;7:601–13.10.1517/17530059.2013.846323Search in Google Scholar PubMed

7. Meroni PL, Chan EK, Damoiseaux J, Andrade LEC, Bossuyt X, Conrad K, et al. Unending story of the indirect immunofluorescence assay on HEp-2 cells: old problems and new solutions? Ann Rheum Dis 2018. DOI: 10.1136/annrheumdis-2018-213440.Search in Google Scholar PubMed

8. Agmon-Levin N, Damoiseaux J, Kallenberg C, Sack U, Witte T, Herold M, et al. International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies. Ann Rheum Dis 2014;73:17–23.10.1136/annrheumdis-2013-203863Search in Google Scholar PubMed

9. Meroni PL, Schur PH. ANA screening: an old test with new recommendations. Ann Rheum Dis 2010;69:1420–2.10.1136/ard.2009.127100Search in Google Scholar PubMed

10. Dellavance A, Cruvinel WM, Francescantonio PL, Andrade LE. Antinuclear antibodies. In: Detrick B, Schmitz JL, Hamilton RG, editors. Manual of Molecular and Clinical Laboratory Immunology. Washington: ASM Press, 2016: 843–58.10.1128/9781555818722.ch87Search in Google Scholar

11. Chan EK, Damoiseaux J, Carballo OG, Conrad K, Cruvinel WM, Francescantonio PL, et al. Report of the first international consensus on standardized nomenclature of antinuclear antibody HEp-2 cell patterns 2014-2015. Front Immunol 2015;6:412.10.3389/fimmu.2015.00412Search in Google Scholar PubMed PubMed Central

12. Andrade LE, Klotz W, Herold M, Conrad K, Rönnelid J, Fritzler MJ, et al. International consensus on antinuclear antibody patterns: definition of the AC-29 pattern associated with antibodies to DNA topoisomerase I. Clin Chem Lab Med 2018;56:1783–88.10.1515/cclm-2018-0188Search in Google Scholar PubMed

13. Leuchten N, Hoyer A, Brinks R, Schoels M, Schneider M, Smolen J, et al. Performance of antinuclear antibodies for classifying systemic lupus erythematosus: a systematic literature review and meta-regression of diagnostic data. Arthritis Care Res (Hoboken) 2018;70:428–38.10.1002/acr.23292Search in Google Scholar PubMed

14. Gladman DD, Ibanez D, Urowitz MB. Systemic lupus erythematosus disease activity index 2000. J Rheumatol 2002;29:288–91.Search in Google Scholar

15. Uribe AG, Vilá LM, McGwin G Jr, Sanchez ML, Reveille JD, Allarcón GS. The systemic lupus activity measure-revised, the Mexican Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), and a modified SLEDAI-2K are adequate instruments to measure disease activity in systemic lupus erythematosus. JRheumatol 2004;31:1934–40.Search in Google Scholar

16. Araújo MN, Leser PG, Gabriel Jr A, Assad RL, Atra E. A simple radial immunohemolysis assay for the measurement of functional complement C2 activity. Braz J Med Biol Res 1991;24:49–57.Search in Google Scholar

17. Mariz HA, Sato EI, Barbosa SH, Rodrigues SH, Dellavance A, Andrade LE. 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 2011;63:191–200.10.1002/art.30084Search in Google Scholar PubMed

18. Chabre H, Amoura Z, Piette JC, Godeau P, Bach JF, Koutouzov S. Presence of nucleosome-restricted antibodies in patients with systemic lupus erythematosus. Arthritis Rheum 1995;38:1485–91.10.1002/art.1780381015Search in Google Scholar PubMed

19. Ng KP, Manson JJ, Rahman A, Isenberg DA. Association of antinucleosome antibodies with disease flare in serologically active clinically quiescent patients with systemic lupus erythematosus. Arthritis Rheum 2006;55:900–4.10.1002/art.22356Search in Google Scholar PubMed

20. Li T, Prokopec SD, Morrison S, Lou W, Reich H, Gladman D, et al. Anti-nucleosome antibodies outperform traditional biomarkers as longitudinal indicators of disease activity in systemic lupus erythematosus. Rheumatology (Oxford) 2015;54:449–57.10.1093/rheumatology/keu326Search in Google Scholar PubMed PubMed Central

21. Zhang CQ, Ren L, Gao F, Mu FY, You YQ, Liu YH. Anti-C1q antibodies are associated with systemic lupus erythematosus disease activity and lupus nephritis in northeast of China. Clin Rheumatol 2011;30:967–73.10.1007/s10067-011-1698-1Search in Google Scholar PubMed

22. Skiljevic D, Jeremic I, Nikolic M, Andrejevic S, Sefik-Bukilica M, Stojimirovic B, et al. Serum DNase I activity in systemic lupus erythematosus: correlation with immunoserological markers, the disease activity and organ involvement. Clin Chem Lab Med 2013;51:1083–91.10.1515/cclm-2012-0521Search in Google Scholar PubMed

23. Viallard JF, Bloch-Michel C, Neau-Cransac M, Taupin JL, Garrigue S, Miossec V, et al. HLA-DR expression on lymphocyte subsets as a marker of disease activity in patients with systemic lupus erythematosus. Clin Exp Immunol 2001;125:485–91.10.1046/j.1365-2249.2001.01623.xSearch in Google Scholar PubMed PubMed Central

24. He C, Mao T, Feng Y, Song T, Qin C, Yan R, et al. Anti-CII antibody as a novel indicator to assess disease activity in systemic lupus erythematosus. Lupus 2015;24:1370–6.10.1177/0961203315588970Search in Google Scholar PubMed

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0638).

Received: 2019-06-25
Accepted: 2020-02-09
Published Online: 2020-03-12
Published in Print: 2020-07-28

©2020 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2019-0638
Keywords for this article
antinuclear antibody; autoantibodies; disease activity; HEp-2 cells; SLEDAI2K; systemic lupus erythematosus

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Quality controls for serology: an unfinished agenda
  4. A modern and pragmatic definition of Laboratory Medicine
  5. Reviews
  6. Blood biochemical characteristics of patients with coronavirus disease 2019 (COVID-19): a systemic review and meta-analysis
  7. ISO/TS 20914:2019 – a critical commentary
  8. Mini Review
  9. Reporting of D-dimer data in COVID-19: some confusion and potential for misinformation
  10. Opinion Paper
  11. Implementation of metrological traceability in laboratory medicine: where we are and what is missing
  12. IFCC Recommendation
  13. Recommendation for performance verification of patient-based real-time quality control
  14. Genetics and Molecular Diagnostics
  15. Comparison of BCR-ABL1 quantification in peripheral blood and bone marrow using an International Scale-standardized assay for assessment of deep molecular response in chronic myeloid leukemia
  16. General Clinical Chemistry and Laboratory Medicine
  17. Risk assessment of the total testing process based on quality indicators with the Sigma metrics
  18. Determination of hemolysis cut-offs for biochemical and immunochemical analytes according to their value
  19. A computer model for professional competence assessment according to ISO 15189
  20. Traceability validation of six enzyme measurements on the Abbott Alinity c analytical system
  21. Evaluating the need for free glycerol blanking for serum triglyceride measurements at Charlotte Maxeke Johannesburg Academic Hospital
  22. Challenges of LC-MS/MS ethyl glucuronide analysis in abstinence monitoring of liver transplant candidates
  23. Changes in the result of antinuclear antibody immunofluorescence assay on HEp-2 cells reflect disease activity status in systemic lupus erythematosus
  24. Reference Values and Biological Variations
  25. Long-term biological variation estimates of 13 hematological parameters in healthy Chinese subjects
  26. Age-specific reference values improve the diagnostic performance of AMH in polycystic ovary syndrome
  27. Establishment of reference intervals for immunoassay analytes of adult population in Saudi Arabia
  28. Hematology and Coagulation
  29. Total haemoglobin – a reference measuring system for improvement of standardisation
  30. Laboratory testing for activated protein C resistance: rivaroxaban induced interference and a comparative evaluation of andexanet alfa and DOAC Stop to neutralise interference
  31. Cancer Diagnostics
  32. Identification of a four-gene methylation biomarker panel in high-grade serous ovarian carcinoma
  33. Performance comparison of two next-generation sequencing panels to detect actionable mutations in cell-free DNA in cancer patients
  34. Diabetes
  35. Availability and analytical quality of hemoglobin A1c point-of-care testing in general practitioners’ offices are associated with better glycemic control in type 2 diabetes
  36. Infectious Diseases
  37. Validation of a chemiluminescent assay for specific SARS-CoV-2 antibody
  38. Dynamic profile and clinical implications of hematological parameters in hospitalized patients with coronavirus disease 2019
  39. Does a change in quality control results influence the sensitivity of an anti-HCV test?
  40. Letters to the Editor
  41. Variability between testing methods for SARS-CoV-2 nucleic acid detection 16 days post-discharge: a case report
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  44. Counting and reporting band count is unreliable practice due to the high inter-observer variability
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  46. How reliable is the detection of anti-mitochondrial antibodies on murine triple-tissue?
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