The Effects of Immunocytochemistry on Diagnostic Accuracy in Thyroid Fine Needle Aspiration Cytology: A Retrospective Study

Year 2025, Volume: 15 Issue: 3, 531 - 537, 30.09.2025

Ali Mızrak , Cansu Benli Işık , Esen Gül Uzuner , Saime Gül Barut

https://doi.org/10.33808/clinexphealthsci.1552921

Abstract

Objective: In thyroid cytology, ancillary studies are often utilized for intermediate-category cases to better differentiate between benign and malignant lesion. For this purpose, immunocytochemical markers may be preferred because they are cheaper than molecular studies and can be applied in many laboratories. This study retrospectively compares the diagnostic accuracy of cytology samples with and without immunocytochemistry and calculates the frequency of immunocytochemical marker use, as well as sensitivity and specificity rates.
Methods: Between 2015 and 2023, 1816 samples from 1506 patients with a histological diagnosis after cytological examination (thyroidectomy-lobectomy) were included. Cases without a histological diagnosis were excluded. The Thin Prep® method was used for all cytological sample preparations, and cell blocks were obtained. Demographic information, Bethesda system categories, immunocytochemical markers used, and histological diagnoses were recorded. Cases using at least one immunocytochemical marker were re-evaluated, and staining results were categorized as positive, focally positive, or negative. SPSS 15® software was used to assess data normality and perform statistical analyses.
Results: The most frequently used markers were HBME-1 (n=167), CK 19 (n=106), Galectin-3 (n=75), and CD 56 (n=6). Sensitivity rates for HBME-1, CK 19, Galectin-3, and CD 56 were 91%, 94%, 76%, and 75%, respectively; specificity rates were 63%, 61%, 80%, and 50%, respectively. Comparing groups with and without immunocytochemistry, the risk of malignancy was: 6.95%-6.97% for Bethesda category II; 21.7%-19.0% for Bethesda category III; 76.0%-37.1% for Bethesda category IV; 94.0%-95.0% for category V; and 100% for category VI.
Conclusion: In the follicular neoplasm group (Bethesda category IV) the risk of malignancy was higher in the immunohistochemistry applied group. No significant difference in malignancy risk was observed between groups with and without immunocytochemistry in other categories. Considering that immunocytochemical markers were predominantly applied in diagnostically challenging Bethesda categories, the similar malignancy risks across groups may suggest immunocytochemistry aids in accurate categorization. However, according to the results of this study, routine use of immunohistochemical markers in thyroid cytology is unnecessary except for Bethesda category IV.

Keywords

CK 19 , Galectin-3 , HBME-1 , Immunocytochemistry , Thyroid

Ethical Statement

This study was approved by Ethics Committee of Haseki Training and Research Hospital, Noninvasive Clinic Ethics Committee (Approval date: 23.05.2024.; Number:26-2024)

Supporting Institution

None

References

• Ali SZ, Baloch ZW, Cochand-Priollet B, Schmitt FC, Vielh P, VanderLaan PA. The 2023 Bethesda system for reporting thyroid cytopathology. Thyroid. 2023;33(9):1039–1044. https://doi.org/10.1089/thy.2023.0141
• Rossi ED, Adeniran AJ, Faquin WC. Pitfalls in thyroid cytopathology. Surg Pathol Clin. 2019;12(4):865–881. https://doi.org/10.1016/j.path.2019.08.001
• Zhu Y, Song Y, Xu G, Fan Z, Ren W. Causes of misdiagnoses by thyroid fine-needle aspiration cytology (FNAC): Our experience and a systematic review. Diagn Pathol. 2020;15(1):1. https://doi.org/10.1186/s13000-019-0924-z
• Cha H, Pyo JY, Hong SW. The usefulness of immunocytochemistry of CD56 in determining malignancy from indeterminate thyroid fine-needle aspiration cytology. J Pathol Transl Med. 2018;52(6):404–410. https://doi.org/10.4132/jptm.2018.09.20
• Xiong Y, Li X, Liang L, Li D, Yan L, Li X, Di J, Li T. Application of biomarkers in the diagnosis of uncertain samples of core needle biopsy of thyroid nodules.Virchows Arch. 2021;479(5):961–974. https://doi.org/10.1007/s00428-021-03161-y.
• Margari N, Giovannopoulos I, Pouliakis A, Mastorakis E, Gouloumi AR, Panayiotides IG, Karakitsos P. Application of immunocytochemistry on cell block sections for the investigation of thyroid lesions. Acta Cytol. 2018;62(2):137–144. https://doi.org/10.115910.1159/000485824
• Cazzaniga G, Seminati D, Smith A, Piga I, Capitoli G, Garancini M, L'Imperio V, Fusco N, Pagni F. Lights on HBME-1: The elusive biomarker in thyroid cancer pathology. J Clin Pathol. 2022;75(9):588–592. https://doi.org/10.1136/jclinpath-2022-208248
• Chuang HW, Wang JS, Tsai JW, Hsu CT, Lin KJ. Immunohistochemistry helps to distinguish noninvasive follicular thyroid neoplasm with papillary-like nuclear features/noninvasive encapsulated follicular variant of papillary thyroid carcinoma with other follicular thyroid lesions. Medicina. 2021; 57:1246. https://doi.org/10.3390/medicina57111246
• Song S, Kim H, Ahn SH. Role of immunohistochemistry in fine needle aspiration and core needle biopsy of thyroid nodules. Clin Exp Otorhinolaryngol. 2019;12(2):224–230. https://doi.org/10.21053/ceo.2018.01011
• Xin Y, Guan D, Meng K, Lv Z, Chen B. Diagnostic accuracy of CK-19, Galectin-3 and HBME-1 on papillary thyroid carcinoma: a meta-analysis. Int J Clin Exp Pathol. 2017;10(8):8130–8140. PMID: 31966665 PMCID: PMC6965469
• Li J, Vasilyeva E, Wiseman SM. Beyond immunohistochemistry and immunocytochemistry: a current perspective on galectin-3 and thyroid cancer. Expert Rev Anticancer Ther. 2019;19(12):1017–1027. https://doi.org/10.1080/14737140.2019.1693270
• Cho U, Kim Y, Jeon S, Jung CK. CD56 expression in papillary thyroid carcinoma is highly dependent on the histologic subtype: A potential diagnostic pitfall. Appl Immunohistochem Mol Morphol. 2022;30(5):389–396. https://doi.org/10.1097/PAI.0000000000001015
• Ahmed HG, El Hag ABM, Alanazi KK, Alkwai HM, Abdrhman AMA, Hassan AOA, Ginawi IAM, Elasbali AM, Sherfi H. Patterns of thyroid tumors in Northern Saudi Arabia with a specific focus on CK19, CD56, and Galectin-3 tumor markers. Eur Rev Med Pharmacol Sci. 2023;27(12):5654–5661. https://doi.org/10.26355/eurrev_202306_32805
• Dixit S, Diwaker P, Wadhwa N, Arora VK. Galectin-3 and CD117 immunocytochemistry in the diagnosis of indeterminate thyroid lesions: A pilot study. Diagn Cytopathol. 2021;49(1):1–9. https://doi.org/10.1002/dc.24835.
• Swan KZ, Madsen SH, Bonnema SJ, Nielsen VE, Jespersen ML. Preoperative BRAFV600E mutation detection in thyroid carcinoma by immunocytochemistry. Apmis. 2022;130(11):627–636. https://doi.org/10.1111/apm.13267.
• Nikas IP, Kazamias G, Vrontaki M, Rapti AS, Mastorakis E. Medullary thyroid carcinoma diagnosed with liquid-based cytology and immunocytochemistry. J Immunoassay Immunochem. 2022;43(5):502–515. https://doi.org/ 10.1080/15321819.2022.2070025
• Liu CY, Bychkov A, Agarwal S, Zhu Y, Hang JF, Lai CR, Na HY, Li W, Liu Z, Jain D, Suzuki A, Hirokawa M, Chia N, Nga ME, Jitpasutham T, Keelawat S, Park SY, Satoh S, Chen CC, Gunawardena D, Kumarasinghe P, Jung CK, Kakudo K. Cytologic diagnosis of medullary thyroid carcinoma in the Asia-Pacific region. Diagn Cytopathol. 2021;49(1):60–69. https://doi.org/10.1002/dc.24586.
• Baloch Z, Mete O, Asa SL. Immunohistochemical biomarkers in thyroid pathology. Endocr Pathol. 2018;29(2):91–112. https://doi.org/10.1007/s12022-018-9532-9
• He SR, Liu LT, Chen RM, Wang MG, Hu ST, Miao G, Chen L, Liu DG. Diagnostic values of nuclear score combined with cyclin D1 immunocytochemistry in indeterminate thyroid follicular nodules in preoperative fine needle aspiration. Zhonghua Bing Li Xue Za Zhi. 2023;52(7):696–701. https://doi.org/10.3760/cma.j.cn112151-20230116-00046
• Monteiro I, Missiaglia E, Sciarra A, Santos JV, Bouilly J, Romero P, Sempoux C, de Leval L. CD73 expression in normal, hyperplastic, and neoplastic thyroid: a systematic evaluation revealing CD73 overexpression as a feature of papillary carcinomas. Virchows Arch. 2021;479(1):209–214. https://doi.org/10.1007/s00428-021-03100-x
• Czajkowski M, Kaemmerer D, Sänger J, Sauter G, Wirtz RM, Schulz S, Lupp A. Comparative evaluation of somatostatin and CXCR4 receptor expression in different types of thyroid carcinoma using well-characterised monoclonal antibodies. Bmc Cancer. 2022;22(1):740. https://doi.org/10.1186/s12885-022-09839-z
• Wang G, Li HN, Cui XQ, Xu T, Dong ML, Li SY, Li XR. S100A1 is a potential biomarker for papillary thyroid carcinoma diagnosis and prognosis. J Cancer. 2021;12(19):5760–5771. https://doi.org/0.7150/jca.51855
• Martins MB, de Assis Batista F, Marcello MA, Bufalo NE, Peres KC, Morari EC, Soares FA, Vassallo J, Ward LS. Clinical utility of the immunohistochemical co-expression of p53 and MDM2 in thyroid follicular lesions. Ann Diagn Pathol. 2021;53:151766. https://doi.org/10.1016/j.anndiagpath.2021.151766
• Ibrahiem AT, Makhdoom AK, Alanazi KS, Alanazi AM, Mukhlef AM, Elshafey SH, Toraih EA, Fawzy MS. Analysis of anti-apoptotic PVT1 oncogene and apoptosis-related proteins (p53, Bcl2, PD-1, and PD-L1) expression in thyroid carcinoma. J Clin Lab Anal. 2022;36(5):e24390. https://doi.org/10.1002/jcla.24390
• Kusafuka K, Yamashita M, Iwasaki T, Tsuchiya C, Kubota A, Hirata K, Murakami A, Muramatsu A, Arai K, Suzuki M. Periostin expression and its supposed roles in benign and malignant thyroid nodules: an immunohistochemical study of 105 cases. Diagn Pathol. 2021;16(1):86. https://doi.org/10.1186/s13000-021-01146-8
• Crescenzi A, Baloch Z. Immunohistochemistry in the pathologic diagnosis and management of thyroid neoplasms. Front Endocrinol (Lausanne). 2023;14:1198099. https://doi.org/10.3389/fendo.2023.1198099
• Kim K, Bae JS, Kim JS, Jung SL, Jung CK. Diagnostic performance of thyroid core needle biopsy using the revised reporting system: Comparison with fine needle aspiration cytology. Endocrinol Metab (Seoul). 2022;37(1):159–169. https://doi.org/doi: 10.3803/EnM.2021.1299
• Kwon H, Lee J, Hong SW, Kwon HJ, Kwak JY, Yoon JH. Fine needle aspiration cytology vs. core needle biopsy for thyroid nodules: a prospective, experimental study using surgical specimen. Taehan Yongsang Uihakhoe Chi. 2022;83(3):645–657. https://doi.org/10.3348/jksr.2021.0125
• Tastekin E, Canberk S, Schmitt FC. Follicular growth pattern disease on thyroid fine-needle aspiration biopsy. Balkan Med J. 2022;39(4):230–238. https://doi.org/ 10.4274/balkanmedj.galenos.2022.2022-5-17
• Alzumaili BA, Krumeich LN, Collins R, Kravchenko T, Ababneh EI, Fisch AS, Faquin WC, Nosé V, Martinez-Lage M, Randolph GW, Gartland RM, Lubitz CC, Sadow PM. A comprehensive study on the diagnosis and management of noninvasive follicular thyroid neoplasm with papillary-like nuclear features. Thyroid. 2023;33(5):566–577. https://doi.org/10.1089/thy.2023.0035