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Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?

Year 2025, Volume: 8 Issue: 4, 139 - 144, 15.07.2025
https://doi.org/10.19127/bshealthscience.1623545

Abstract

The indirect immunofluorescence (IIF) method for anti-nuclear antibody (ANA) testing is pivotal in the diagnosis of autoimmune diseases. Despite extensive research on nuclear staining patterns, cytoplasmic and mitotic patterns remain less understood. This study retrospectively analyzed 12,674 ANA test results from a tertiary medical microbiology laboratory over three years to assess the prevalence and diagnostic implications of these patterns. ANA positivity was observed in 24.2% of samples, with cytoplasmic and mitotic patterns accounting for 9.2%. Notably, these patterns were predominantly found in ANA-negative samples, with intercellular bridge (AC-27) emerging as the most frequent pattern. While a substantial proportion of cytoplasmic and mitotic patterns were detected among rheumatology patients, no significant correlation was identified between specific patterns and autoimmune diagnoses. These findings underscore the importance of consistent reporting of cytoplasmic and mitotic patterns, as recommended by the International Consensus on ANA Patterns (ICAP). The incorporation of these patterns into routine diagnostic reports has the potential to enhance diagnostic accuracy, particularly in cases ANA-negative. Further research is essential to elucidate their clinical significance and optimize laboratory practices.

Ethical Statement

The study was conducted in accordance with the principles of the Declaration of Helsinki. Clinical Research Ethics Committee of Afyonkarahisar Health Sciences University approved this study (approval date: June 02, 2023, protocol code: 258).

Project Number

-

References

  • Aringer M, Costenbader K, Daikh D, Brinks R, Mosca M, Ramsey Goldman R, Smolen JS, Wofsy D, Boumpas DT, Kamen DL, Jayne D, Cervera R, Costedoat Chalumeau N, Diamond B, Gladman DD, Hahn BH, Hiepe F, Jacobsen S, Khanna D, Lerstrøm K, Massarotti E, McCune W, Ruiz Irastorza G, Sanchez Guerrero J, Schneider M, Urowitz MB, Bertsias G, Hoyer BF, Leuchten N, Tani C, Tedeschi SK, Touma Z, Schmajuk G, Anić B, Assan F, Chan TM, Clarke AE, Crow MK, Czirják L, Doria A, Graninger W, Halda Kiss B, Hasni S, Izmirly PM, Jung M, Kumánovics G, Mariette X, Padjen I, Pego Reigosa JM, Romero Díaz J, Rúa Figueroa I, Seror R, Stummvoll GH, Tanaka Y, Tektonidou MG, Vasconcelos C, Vital EM, Wallace DJ, Yavuz S, Meroni PL, Fritzler MJ, Naden RP, Dörner T, Johnson SR. 2019. European League Against Rheumatism/American College of Rheumatology classification criteria for systemic lupus erythematosus. Arthritis Rheumatol, 71: 1400-1412.
  • Betancur JF, Londoño A, Estrada VE, Matteson EL, Rojas Villarraga A, Anaya JM. 2018. Uncommon patterns of antinuclear antibodies recognizing mitotic spindle apparatus antigens and clinical associations. Medicine, 97: e11727.
  • Bilgin M, Baklacıoğlu Ş. 2023. İndirekt immünofloresan yöntemiyle çalışılan antinükleer antikor sonuçlarının değerlendirilmesi. Turk Mikrobiyol Cemiy Derg, 53(3): 143-148.
  • Brito Fde A, Santos SM, Ferreira GA, Pedrosa W, Gradisse J, Costa LC, Neves SP. 2014. Detection of anti nuclear antibodies by indirect immunofluorescence on HEp 2 cells: setting the appropriate screening dilution for the diagnosis of autoimmune rheumatic diseases. Rev Bras Reumatol, 54(1): 13-20.
  • Chan EK, Damoiseaux J, Carballo OG, Conrad K, de Melo Cruvinel W, Francescantonio PL, Fritzler MJ, Garcia De La Torre I, Herold M, Klotz W. 2015. Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp 2 Cell Patterns 2014–2015. Front Immunol, 6: 412.
  • Chan EK, Damoiseaux J, de Melo Cruvinel W, Carballo OG, Conrad K, Francescantonio PL, Fritzler MJ, Garcia De La Torre I, Herold M, Klotz W. 2015. Report on the Second International Consensus on ANA Pattern (ICAP) Workshop in Dresden. Lupus, 25: 797-804.
  • Damoiseaux J, Andrade LEC, Carballo OG, Conrad K, Francescantonio PLC, Fritzler MJ. 2019. Clinical relevance of HEp 2 indirect immunofluorescent patterns: the International Consensus on ANA patterns (ICAP) perspective. Ann Rheum Dis, 78(7): 879-889.
  • Damoiseaux J, von Mühlen CA, Garcia De La Torre I, Carballo OG, de Melo Cruvinel W, Francescantonio PL, Fritzler MJ, Herold M, Klotz W, Andrade LE. 2016. International consensus on ANA patterns (ICAP): The bumpy road towards a consensus on reporting ANA results. Auto Immun Highlights, 7(1): 1.
  • Dinse GE, Parks CG, Weinberg CR, Co CA, Wilkerson J, Zeldin DC, Miller FW. 2020. Increasing prevalence of antinuclear antibodies in the United States. Arthritis Rheumatol, 72(6): 1026-1035.
  • Infantino M, Palterer B, Biagiotti R, Veneri D, Li Gobbi F, Benucci M. 2018. Reflex testing of speckled cytoplasmic patterns observed in routine ANA HEp 2 indirect immunofluorescence with a multiplex anti synthetase dot blot assay: a multicentric pilot study. Immunol Res, 66(1): 74-78.
  • Kasifoglu N, Yasar Bilge NS, Kasifoglu T. 2022. The anti cytoplasmic and anti mitotic autoantibodies; Are these antibodies associated with diseases? Osmangazi J Med, 44(6): 851-861.
  • Klimud. 2020. Otoantikorların Laboratuvar Tanısı Rehberi. Ankara, Klimud, Türkiye, pp: 25.
  • Lerner A, Jeremias P, Matthias T. 2015. The world incidence and prevalence of autoimmune diseases is increasing. Int J Celiac Dis, 3(4): 151-155.
  • Mahler M, Andrade LE, Casiano CA, Malyavantham K, Fritzler MJ. 2019. Implications for redefining the dense fine speckled and related indirect immunofluorescence patterns. Expert Rev Clin Immunol, 15(5): 447-448.
  • Mahler M, Pierangeli S, Meroni PL, Fritzler MJ. 2014. Autoantibodies in systemic autoimmune disorders. J Immunol Res, 2014: 263091.
  • Meroni PL, Schur PH. 2010. ANA screening: an old test with new recommendations. Ann Rheum Dis, 69(8): 1420-1422.
  • Nanda R, Gupta P, Patel S, Shah S, Mohapatra E. 2021. Uncommon antinuclear antibody patterns as diagnostic indicators. Clin Biochem, 90: 28-33.
  • Pascual V, Perez I, Sánchez Ramón S, Khamashta M, Caballero T, Rodriguez Mahou M. 2015. Antibodies to mitotic apparatus: new association with cholestatic liver disease. Am J Gastroenterol, 110: 1736-1737.
  • Staruszkiewicz M, Pituch Noworolska A, Skoczen S. 2023. Uncommon types of autoantibodies – Detection and clinical associations. Autoimmun Rev, 22(3): 103263.
  • Stinton LM, Eystathioy T, Selak S, Chan EKL, Fritzler MJ. 2004. Autoantibodies to protein transport and messenger RNA processing pathways: endosomes, lysosomes, Golgi complex, proteasomes, assemblyosomes, exosomes, and GW bodies. Clin Immunol, 110(1): 30-44.
  • Szalat R, Ghillani Dalbin P, Jallouli M, Le Thi Huong D, Andreu MR, Leroux G. 2010. Anti NuMA1 and anti NuMA2 (anti HsEg5) antibodies: clinical and Immunological features: a propos of 40 new cases and review of the literature. Autoimmun Rev, 9: 652-656.
  • Vermeersch P, Bossuyt X. 2013. Prevalence and clinical significance of rare antinuclear antibody patterns. Autoimmun Rev, 12(10): 998-1003.
  • von Mühlen CA, Garcia De La Torre I, Infantino M, Conrad K, Damoiseaux J, Fritzler MJ. 2021. How to report the antinuclear antibodies (anti-cell antibodies) test on HEp-2 cells: guidelines from the ICAP initiative. Immunol Res, 69(6): 594-608.
  • Wiik AS, Høier-Madsen M, Forslid J, Charles P, Meyrowitsch J. 2010. Antinuclear antibodies: a contemporary nomenclature using HEp-2 cells. J Autoimmun, 35(3): 276-290.

Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?

Year 2025, Volume: 8 Issue: 4, 139 - 144, 15.07.2025
https://doi.org/10.19127/bshealthscience.1623545

Abstract

The indirect immunofluorescence (IIF) method for anti-nuclear antibody (ANA) testing is pivotal in the diagnosis of autoimmune diseases. Despite extensive research on nuclear staining patterns, cytoplasmic and mitotic patterns remain less understood. This study retrospectively analyzed 12,674 ANA test results from a tertiary medical microbiology laboratory over three years to assess the prevalence and diagnostic implications of these patterns. ANA positivity was observed in 24.2% of samples, with cytoplasmic and mitotic patterns accounting for 9.2%. Notably, these patterns were predominantly found in ANA-negative samples, with intercellular bridge (AC-27) emerging as the most frequent pattern. While a substantial proportion of cytoplasmic and mitotic patterns were detected among rheumatology patients, no significant correlation was identified between specific patterns and autoimmune diagnoses. These findings underscore the importance of consistent reporting of cytoplasmic and mitotic patterns, as recommended by the International Consensus on ANA Patterns (ICAP). The incorporation of these patterns into routine diagnostic reports has the potential to enhance diagnostic accuracy, particularly in cases ANA-negative. Further research is essential to elucidate their clinical significance and optimize laboratory practices.

Ethical Statement

The study was conducted in accordance with the principles of the Declaration of Helsinki. Clinical Research Ethics Committee of Afyonkarahisar Health Sciences University approved this study (approval date: June 02, 2023, protocol code: 258).

Supporting Institution

None

Project Number

-

Thanks

None

References

  • Aringer M, Costenbader K, Daikh D, Brinks R, Mosca M, Ramsey Goldman R, Smolen JS, Wofsy D, Boumpas DT, Kamen DL, Jayne D, Cervera R, Costedoat Chalumeau N, Diamond B, Gladman DD, Hahn BH, Hiepe F, Jacobsen S, Khanna D, Lerstrøm K, Massarotti E, McCune W, Ruiz Irastorza G, Sanchez Guerrero J, Schneider M, Urowitz MB, Bertsias G, Hoyer BF, Leuchten N, Tani C, Tedeschi SK, Touma Z, Schmajuk G, Anić B, Assan F, Chan TM, Clarke AE, Crow MK, Czirják L, Doria A, Graninger W, Halda Kiss B, Hasni S, Izmirly PM, Jung M, Kumánovics G, Mariette X, Padjen I, Pego Reigosa JM, Romero Díaz J, Rúa Figueroa I, Seror R, Stummvoll GH, Tanaka Y, Tektonidou MG, Vasconcelos C, Vital EM, Wallace DJ, Yavuz S, Meroni PL, Fritzler MJ, Naden RP, Dörner T, Johnson SR. 2019. European League Against Rheumatism/American College of Rheumatology classification criteria for systemic lupus erythematosus. Arthritis Rheumatol, 71: 1400-1412.
  • Betancur JF, Londoño A, Estrada VE, Matteson EL, Rojas Villarraga A, Anaya JM. 2018. Uncommon patterns of antinuclear antibodies recognizing mitotic spindle apparatus antigens and clinical associations. Medicine, 97: e11727.
  • Bilgin M, Baklacıoğlu Ş. 2023. İndirekt immünofloresan yöntemiyle çalışılan antinükleer antikor sonuçlarının değerlendirilmesi. Turk Mikrobiyol Cemiy Derg, 53(3): 143-148.
  • Brito Fde A, Santos SM, Ferreira GA, Pedrosa W, Gradisse J, Costa LC, Neves SP. 2014. Detection of anti nuclear antibodies by indirect immunofluorescence on HEp 2 cells: setting the appropriate screening dilution for the diagnosis of autoimmune rheumatic diseases. Rev Bras Reumatol, 54(1): 13-20.
  • Chan EK, Damoiseaux J, Carballo OG, Conrad K, de Melo Cruvinel W, Francescantonio PL, Fritzler MJ, Garcia De La Torre I, Herold M, Klotz W. 2015. Report of the First International Consensus on Standardized Nomenclature of Antinuclear Antibody HEp 2 Cell Patterns 2014–2015. Front Immunol, 6: 412.
  • Chan EK, Damoiseaux J, de Melo Cruvinel W, Carballo OG, Conrad K, Francescantonio PL, Fritzler MJ, Garcia De La Torre I, Herold M, Klotz W. 2015. Report on the Second International Consensus on ANA Pattern (ICAP) Workshop in Dresden. Lupus, 25: 797-804.
  • Damoiseaux J, Andrade LEC, Carballo OG, Conrad K, Francescantonio PLC, Fritzler MJ. 2019. Clinical relevance of HEp 2 indirect immunofluorescent patterns: the International Consensus on ANA patterns (ICAP) perspective. Ann Rheum Dis, 78(7): 879-889.
  • Damoiseaux J, von Mühlen CA, Garcia De La Torre I, Carballo OG, de Melo Cruvinel W, Francescantonio PL, Fritzler MJ, Herold M, Klotz W, Andrade LE. 2016. International consensus on ANA patterns (ICAP): The bumpy road towards a consensus on reporting ANA results. Auto Immun Highlights, 7(1): 1.
  • Dinse GE, Parks CG, Weinberg CR, Co CA, Wilkerson J, Zeldin DC, Miller FW. 2020. Increasing prevalence of antinuclear antibodies in the United States. Arthritis Rheumatol, 72(6): 1026-1035.
  • Infantino M, Palterer B, Biagiotti R, Veneri D, Li Gobbi F, Benucci M. 2018. Reflex testing of speckled cytoplasmic patterns observed in routine ANA HEp 2 indirect immunofluorescence with a multiplex anti synthetase dot blot assay: a multicentric pilot study. Immunol Res, 66(1): 74-78.
  • Kasifoglu N, Yasar Bilge NS, Kasifoglu T. 2022. The anti cytoplasmic and anti mitotic autoantibodies; Are these antibodies associated with diseases? Osmangazi J Med, 44(6): 851-861.
  • Klimud. 2020. Otoantikorların Laboratuvar Tanısı Rehberi. Ankara, Klimud, Türkiye, pp: 25.
  • Lerner A, Jeremias P, Matthias T. 2015. The world incidence and prevalence of autoimmune diseases is increasing. Int J Celiac Dis, 3(4): 151-155.
  • Mahler M, Andrade LE, Casiano CA, Malyavantham K, Fritzler MJ. 2019. Implications for redefining the dense fine speckled and related indirect immunofluorescence patterns. Expert Rev Clin Immunol, 15(5): 447-448.
  • Mahler M, Pierangeli S, Meroni PL, Fritzler MJ. 2014. Autoantibodies in systemic autoimmune disorders. J Immunol Res, 2014: 263091.
  • Meroni PL, Schur PH. 2010. ANA screening: an old test with new recommendations. Ann Rheum Dis, 69(8): 1420-1422.
  • Nanda R, Gupta P, Patel S, Shah S, Mohapatra E. 2021. Uncommon antinuclear antibody patterns as diagnostic indicators. Clin Biochem, 90: 28-33.
  • Pascual V, Perez I, Sánchez Ramón S, Khamashta M, Caballero T, Rodriguez Mahou M. 2015. Antibodies to mitotic apparatus: new association with cholestatic liver disease. Am J Gastroenterol, 110: 1736-1737.
  • Staruszkiewicz M, Pituch Noworolska A, Skoczen S. 2023. Uncommon types of autoantibodies – Detection and clinical associations. Autoimmun Rev, 22(3): 103263.
  • Stinton LM, Eystathioy T, Selak S, Chan EKL, Fritzler MJ. 2004. Autoantibodies to protein transport and messenger RNA processing pathways: endosomes, lysosomes, Golgi complex, proteasomes, assemblyosomes, exosomes, and GW bodies. Clin Immunol, 110(1): 30-44.
  • Szalat R, Ghillani Dalbin P, Jallouli M, Le Thi Huong D, Andreu MR, Leroux G. 2010. Anti NuMA1 and anti NuMA2 (anti HsEg5) antibodies: clinical and Immunological features: a propos of 40 new cases and review of the literature. Autoimmun Rev, 9: 652-656.
  • Vermeersch P, Bossuyt X. 2013. Prevalence and clinical significance of rare antinuclear antibody patterns. Autoimmun Rev, 12(10): 998-1003.
  • von Mühlen CA, Garcia De La Torre I, Infantino M, Conrad K, Damoiseaux J, Fritzler MJ. 2021. How to report the antinuclear antibodies (anti-cell antibodies) test on HEp-2 cells: guidelines from the ICAP initiative. Immunol Res, 69(6): 594-608.
  • Wiik AS, Høier-Madsen M, Forslid J, Charles P, Meyrowitsch J. 2010. Antinuclear antibodies: a contemporary nomenclature using HEp-2 cells. J Autoimmun, 35(3): 276-290.
There are 24 citations in total.

Details

Primary Language English
Subjects ​Internal Diseases, Epidemiology (Other)
Journal Section Research Article
Authors

Melahat Gürbüz 0000-0001-6290-1216

Project Number -
Publication Date July 15, 2025
Submission Date January 20, 2025
Acceptance Date May 10, 2025
Published in Issue Year 2025 Volume: 8 Issue: 4

Cite

APA Gürbüz, M. (2025). Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns? Black Sea Journal of Health Science, 8(4), 139-144. https://doi.org/10.19127/bshealthscience.1623545
AMA Gürbüz M. Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns? BSJ Health Sci. July 2025;8(4):139-144. doi:10.19127/bshealthscience.1623545
Chicago Gürbüz, Melahat. “Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?”. Black Sea Journal of Health Science 8, no. 4 (July 2025): 139-44. https://doi.org/10.19127/bshealthscience.1623545.
EndNote Gürbüz M (July 1, 2025) Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns? Black Sea Journal of Health Science 8 4 139–144.
IEEE M. Gürbüz, “Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?”, BSJ Health Sci., vol. 8, no. 4, pp. 139–144, 2025, doi: 10.19127/bshealthscience.1623545.
ISNAD Gürbüz, Melahat. “Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?”. Black Sea Journal of Health Science 8/4 (July2025), 139-144. https://doi.org/10.19127/bshealthscience.1623545.
JAMA Gürbüz M. Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns? BSJ Health Sci. 2025;8:139–144.
MLA Gürbüz, Melahat. “Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns?”. Black Sea Journal of Health Science, vol. 8, no. 4, 2025, pp. 139-44, doi:10.19127/bshealthscience.1623545.
Vancouver Gürbüz M. Evaluation of Hep-2 Cell Patterns: How Often Do We Report Cytoplasmic and Mitotic Patterns? BSJ Health Sci. 2025;8(4):139-44.