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Anti-Ki/anti-PA28γ autoantibodies contribute to the HEp-2 indirect immunofluorescence nuclear speckled pattern

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Veröffentlicht/Copyright: 30. November 2022
Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 61 Heft 3

Abstract

Objectives

Antinuclear antibodies (ANAs) are associated with several autoimmune diseases. Indirect immunofluorescence (IIF) on human epithelial type 2 (HEp-2) cells is the golden standard for ANA detection in the clinic. In case of a positive HEp-2 IIF test result, follow-up tests are done to determine autoantibody specificity. For a fraction of the HEp-2 IIF-positive samples, the nature of the autoantigens remains uncharacterized. Our objective was to characterize autoantigens in such samples.

Methods

To characterize autoantigens in an unbiased way, we combined protein immunoprecipitation with liquid chromatography (LC) tandem mass spectrometry (MS/MS) sequencing.

Results

Using such approach we detected the Ki antigen, also referred to as PA28γ, in the immunoprecipitate of serum samples of three individuals with an autoimmune disease. The HEp-2 nuclear speckled IIF fluorescent signal of all three serum samples was abolished after pre-absorption of the serum with recombinant Ki antigen, confirming that autoantibodies against Ki underlie the HEp-2 IIF signal.

Conclusions

Our data suggest that anti-Ki autoantibodies can underlie a nuclear speckled HEp-2 IIF pattern.

Keywords: autoantibodies; HEp-2 IIF; Ki antigen; proteasome activator 28γ (PA28γ)
Corresponding author: Xavier Bossuyt, Clinical and Diagnostic Immunology, Department of Microbiology, Immunology and Transplantation, KU Leuven, Leuven, Belgium; and Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium, E-mail:

Funding source: KU Leuven

Award Identifier / Grant number: C3 (C3/20/042)

Acknowledgments

We thank SyBioMa, the Proteomics Core of the Biomedical Science Group, KU Leuven. We thank Kusay Arat for his technical assistance and Professor Sebastien Carpentier for the constructive discussions.

  1. Research funding: This study was funded by the Research Fund KU Leuven C3 (C3/20/042).

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

  3. Competing interests: Authors declare no competing interests.

  4. Informed consent: This was a retrospective study using left-over material of samples submitted to the clinical laboratory (secondary use) for which an informed consent waiver was authorized by the ethics committee University Hospital Leuven. Controls gave informed consent (study S63708 Ethics committee UZ Leuven).

  5. Ethical approval: The study was approved by the Ethical Committees of the University Hospitals Leuven (S60347/B32220071204) (Leuven). An MTA/DTA between University Hospitals Leuven and Hôpital Henri Mondor was in place.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2022-0858).

Received: 2022-09-01
Accepted: 2022-11-16
Published Online: 2022-11-30
Published in Print: 2023-02-23

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2022-0858
Schlagwörter für diesen Artikel
autoantibodies; HEp-2 IIF; Ki antigen; proteasome activator 28γ (PA28γ)

Artikel in diesem Heft

  1. Frontmatter
  2. Editorial
  3. Cell population data: much more to explore
  4. Reviews
  5. Differences between high-sensitivity cardiac troponin T and I in stable populations: underlying causes and clinical implications
  6. Choosing which in-hospital laboratory tests to target for intervention: a scoping review
  7. Opinion Papers
  8. Definitions and major prerequisites of direct and indirect approaches for estimating reference limits
  9. An algorithm for PCT-guided antimicrobial therapy: a consensus statement by Japanese experts
  10. General Clinical Chemistry and Laboratory Medicine
  11. The usefulness of implementing minimum retest intervals in reducing inappropriate laboratory test requests in a Dutch hospital
  12. Using three external quality assurance schemes to achieve equivalent international normalized ratio results in primary and secondary healthcare
  13. Optimizing the screening of alpha-1 antitrypsin deficiency using serum protein electrophoresis
  14. Anti-Ki/anti-PA28γ autoantibodies contribute to the HEp-2 indirect immunofluorescence nuclear speckled pattern
  15. Simultaneous quantification of tryptophan metabolites by liquid chromatography tandem mass spectrometry during early human pregnancy
  16. Reference Values and Biological Variations
  17. Verification of sex- and age-specific reference intervals for 13 serum steroids determined by mass spectrometry: evaluation of an indirect statistical approach
  18. Cancer Diagnostics
  19. Mucin 13 (MUC13) as a candidate biomarker for ovarian cancer detection: potential to complement CA125 in detecting non-serous subtypes
  20. Increased levels of N6-methyladenosine in peripheral blood RNA: a perspective diagnostic biomarker and therapeutic target for non-small cell lung cancer
  21. Cardiovascular Diseases
  22. Diagnostic utility of total NT-proBNP testing by immunoassay based on antibodies targeting glycosylation-free regions of NT-proBNP
  23. Infectious Diseases
  24. Serial measurement of circulating calprotectin as a prognostic biomarker in COVID-19 patients in intensive care setting
  25. Evaluation of ichroma™ COVID-19 interferon gamma release assay for detection of vaccine-induced immunity in healthcare workers
  26. Application of ultrasensitive assay for SARS-CoV-2 antigen in nasopharynx in the management of COVID-19 patients with comorbidities during the peak of 2022 Shanghai epidemics in a tertiary hospital
  27. Developing a machine learning prediction algorithm for early differentiation of urosepsis from urinary tract infection
  28. Letters to the Editor
  29. A panhaemocytometric approach to COVID-19: the importance of cell population data on Sysmex XN-series analysers in severe disease
  30. Critical appraisal of “choosing which in-hospital laboratory tests to target for intervention: a scoping review”
  31. What is the best external quality control sample for your laboratory?
  32. Pre-analytical variability of the Lumipulse immunoassay for plasma biomarkers of Alzheimer’s disease
  33. Biological variation of serum iron from the European biological variation study (EuBIVAS)
  34. Tube shaking and pneumatic transportation: impact on presepsin concentrations measured by both fully automated and POCT analyzers
  35. Endogenous isobaric interference on serum 17 hydroxyprogesterone by liquid chromatography-tandem mass spectrometry methods
  36. A simulation model for organization and management skills assessment that meets ISO 15189
  37. Congress Abstracts
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