KRONİK LENFOSİTİK LÖSEMİ/KÜÇÜK LENFOSİTİK LENFOMADA KEMİK İLİĞİ AKIM SITOMETRİSİ BULGULARI İLE KEMİK İLİĞİ BIYOPSİSİ İMMÜNOHİSTOKİMYASAL BULGULARININ KARŞILAŞTIRILMASI: METODOLOJİK BİR ÇALIŞMA

Yıl 2023, Cilt: 4 Sayı: 1, 34 - 40, 29.03.2023

Çiğdem Özdemir , Filiz Yavaşoğlu , Yiğit Şenol , Merve Şahin , Tülay Köken

https://doi.org/10.48176/esmj.2023.101

Öz

Giriş: Atipik formları da olan kronik lenfositik lösemi/küçük lenfositik lenfomanın (KLL/SLL) teşhisi çok zor olabilir. Bu çalışmanın amacı erişkinlerde en sık görülen matür B hücreli lösemi/lenfoma olan KLL/SLL'de kemik iliği biyopsisinde immünohistokimyasal (İHK) ve akım sitometrik (AS) immünfenotipleme yöntemlerini karşılaştırmaktır. Bu iki yöntemde KLL/SLL'yi diğer matür B hücreli lösemi/lenfomalardan ayırt edebilecek minimum panelleri belirlemektir.
Yöntem: Çalışmaya matür B hücreli lösemi/lenfoma tanılı 68 erişkin hasta dahil edildi. Kesin tanı periferik yayma, kemik iliği aspirasyonu, kemik iliği biyopsisi, İHK analizi ve AS analizi, klinik ve genetik çalışmalar sonucunda konuldu. Hücre sayıları, kesin tanı ve immünofenotipleme her iki yöntemle karşılaştırıldı. Sitogenik bulgular değerlendirildi. KLL/SLL için İHK ve AS sonuçlarının duyarlılığı, özgüllüğü, pozitif öngörü ve negatif öngörü değerleri hesaplandı.
Bulgular: Olguların %64,7'sinde KLL/SLL, %35,29'unda KLL/SLL dışı lenfomalar saptandı. 68 vakanın ortanca yaşı 66,5 olup, %69,18'i erkek ve %30,82'si kadındı. AS ve İHK’da tanı uyumluluğu %100 ile en yüksek tüylü hücreli lösemi/lenfomada oldu. KLL/SLL'de %90 olarak bulundu. Diğer lenfomalarda kesin tanı ancak biyopsi ile konulabildi. Kemik iliği biyopsisinde CD23+CD43+CD5+LEF-1 ile yapılan immünofenotipleme, KLL/SLL için %95,45 duyarlılık (aralık, %84,53-99,44) ve %100,00 özgüllük (aralık, %85,75-100,00) gösterdi. FC'de CD23+CD43+CD5+CD200+ ile yapılan immünofenotipleme, KLL/SLL için %100,00 duyarlılık (aralık, %91,96-100,00) ve %82,61 özgüllük (aralık, %61,22 -95,05) gösterdi.
Sonuç: Çalışmamız matür b hücreli lösemi/lenfomanın kemik iliği biyopsisinde İHK ve AS'de immünofenotiplemeyi karşılaştıran en kapsamlı çalışMADIR. İHK'da CD23(+), CD43(+), CD5(+) paneline LEF-1'in, AS'de aynı panele CD200'ün eklenmesi klasik tip ve atipik KLL/SLL tanısında ve diğer matür B hücreli lösemi/lenfomalardan ayırıcı tanısında oldukça faydalıdır.

Anahtar Kelimeler

Lenfoma, İmmünohistokimya, Kemik İliği, Akım Sitometri

A COMPARISON OF BONE MARROW FLOW CYTOMETRY FINDINGS AND BONE MARROW BIOPSY İMMUNOHISTOCHEMICAL FINDINGS IN CHRONIC LYMPHOCYTIC LEUKEMIA/SMALL LYMPHOCYTIC LYMPHOMA: A METHODOLOGICAL STUDY

Öz

Introduction: The diagnosis of chronic lymphocytic leukemia/ small lymphocytic lymphoma (CLL/SLL), which also includes atypical forms, can be very difficult. The aim of this study is to
compare immunohistochemical (IHC) and flow cytometric (FC) immunophenotyping methods in the CLL/SLL, the most common mature B-cell leukemia/lymphoma of adults, in bone marrow biopsy. In these two methods, it is to determine the minimum panels that can differentiate CLL/SLL from other mature B-cell leukemia/lymphomas.
Material and Method: The study included 68 adult patients diagnosed with mature B cell leukemia/lymphoma. Final diagnoses were made on the basis of peripheral smear, bone marrow aspirate, bone marrow biopsy, IHC and FC analysis, clinical and genetic studies. Cell counts, final diagnosis and immunophenotyping were compared with both methods. Cytogenic findings were evaluated. The sensitivity, specificity, positive and negative predictive values of IHC and FC results for CLL/SLL were calculated.
Results: CLL/SLL was diagnosed in 64.7% of the cases and non- CLL/SLL in 35.29% of the cases. The median age was 66.50 years, 69.18% male and 30.82% female. The diagnosis compatibility in FC
and IHC was found to be highest in hairy cell leukemia/lymphoma with 100%, and in CLL/SLL with 90%. In other lymphomas, a definitive diagnosis could only be made with biopsy. Immunophenotyping made with CD23+CD43+CD5+LEF-1 in the bone marrow biopsy showed a sensitivity of 95.45% (range, 84.53-99.44%) and a specificity of 100.00% (range, 85.75-100.00%) for CLL/SLL. Immunophenotyping made with CD23+CD43+CD5+CD200+ in FC showed a sensitivity of 100.00% (range, 91.96-100.00%) and a specificity of 82.61% (range, 61.22-95.05%) for CLL/SLL.
Conclusion: This was the most extensive study to have compared immunophenotyping in IHC and FC in bone marrow biopsies of mature B cell leukemia/lymphoma. The addition of LEF-1 to the CD23(+), CD43(+), CD5(+) panel in IHC and the addition of CD200 to the same panel in FC will be extremely helpful in the differential diagnosis of typical/atypical CLL/SLL from mature B cell leukemia/lymphoma.

Anahtar Kelimeler

Lymphoma, Immunohistochemistry, Bone Marrow, Flow Cytometry

Kaynakça

• 1. Swerdlow SH, Campo E, Harris NL, et al. WHO classification of tumours of haematopoietic and lymphoid tissues. 4rd ed. Lyon, International Agency for Research on Cancer; 2017.
• 2. Heel K, Tabone T, Röhrig KJ, et al. Developments in the immunophenotypic analysis of haematological malignancies. Blood Rev 2013;27:193-207.
• 3. Wolach O, Fraser A, Luchiansky M, et al. Can flow cytometry of bone marrow aspirate predict outcome of patients with diffuse large B cell lymphoma? A retrospective single centre study. Hematol Oncol 2015;33:42-7.
• 4. Sovani V, Harvey C, Haynes AP, McMillan AK, Clark DM, O’Connor SR. Bone marrow trephine biopsy involvement by lymphoma: review of histopathological features in 511 specimens and correlation with diagnostic biopsy, aspirate and peripheral blood findings. J Clin Pathol 2014;67:389-95.
• 5. Abdel-Ghafar AA-A, El Din El Telbany MAS, Mahmoud HM, El-Sakhawy YN. Immunophenotyping of chronic B-cell neoplasms: flow cytometry versus immunohistochemistry. Hematology Reports 2012;4:e3.
• 6. Soliman DS, Al‐Kuwari E, Siveen KS, et al. Downregulation of lymphoid enhancer‐binding factor 1 (LEF‐1) expression (by immunohistochemistry and/flow cytometry) in chronic lymphocytic leukemia with atypical immunophenotypic and cytologic features. International Journal of Laboratory Hematology 2021;43:515-25.
• 7. Dewan K, Mann N, Chatterjee T. Comparing flow cytometry immunophenotypic and immunohistochemical analyses in diagnosis and prognosis of chronic lymphoproliferative disorders: Experience from a Tertiary Care Center. Clinical Cancer Investigation Journal 2015;4:707-12.
• 8. Marti GE, Rawstron AC, Ghia P, et al. Diagnostic criteria for monoclonal B‐cell lymphocytosis. Br J Haematol 2005;130:325- 32.
• 9. Habboushe J. Diagnostic test evaluation calculator. Available at: https://www.medcalc.org/calc/diagnostic_test.php (Version 20.106). Accessed April 26, 2022.
• 10. Mhawech P, Buffone GJ, Khan SP, Gresik MV. Cytochemical staining and flow cytometry methods applied to the diagnosis of acute leukemia in the pediatric population: an assessment of relative usefulness. J Pediatr Hematol Oncol 2001;23:89-92.
• 11. Landry M, Bienz M-N, Sawan B, et al. Bone marrow immunohistochemistry and flow cytometry in the diagnosis of malignant hematologic diseases with emphasis on lymphomas: A comparative retrospective study. Applied Immunohistochemistry & Molecular Morphology 2020;28:508-12.
• 12. Merli M, Arcaini L, Boveri E, et al. Assessment of bone marrow involvement in non‐Hodgkin’s lymphomas: comparison between histology and flow cytometry. Eur J Haematol. 2010;85:405-15.
• 13. Brestoff JR, Frater JL. Contemporary challenges in clinical flow cytometry: small samples, big data, little time. The Journal of Applied Laboratory Medicine 2022;7:931-44.
• 14. Menter T, Dirnhofer S, Tzankov A. LEF1: a highly specific marker for the diagnosis of chronic lymphocytic B cell leukaemia/ small lymphocytic B cell lymphoma. J Clin Pathol 2015;68:473-8.
• 15. Köhnke T, Wittmann VK, Bücklein VL, et al. Diagnosis of CLL revisited: increased specificity by a modified five‐marker scoring system including CD 200. Br J Haematol 2017;179:480-7.
• 16. Campo E, Jaffe E. S, Cook J. R, et al . The International Consensus Classification of Mature Lymphoid Neoplasms: a report from the Clinical Advisory Committee. Blood 2022;140:1229-53.
• 17. Alapat D, Coviello-Malle J, Owens R, et al. Diagnostic usefulness and prognostic impact of CD200 expression in lymphoid malignancies and plasma cell myeloma. Am J Clin Pathol 2012;137:93- 100.
• 18. Sandes AF, de Lourdes Chauffaille M, Oliveira CRM, et al. CD200 has an important role in the differential diagnosis of mature B‐cell neoplasms by multiparameter flow cytometry. Cytometry Part B: Clinical Cytometry 2014;86:98-105.
• 19. Matutes E, Owusu-Ankomah K, Morilla R, Houlihan A, Que T, Catovsky D. The immunological profile of B-cell disorders and proposal of a scoring system for the diagnosis of CLL. Leukemia 1994;8:1640-5.
• 20. Rawstron AC, Kreuzer KA, Soosapilla A, et al. Reproducible diagnosis of chronic lymphocytic leukemia by flow cytometry: An European Research Initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation project. Cytometry Part B: Clinical Cytometry 2018;94:121-8.
• 21. Li Y, Tong X, Huang L, et al. A new score including CD43 and CD180: Increased diagnostic value for atypical chronic lymphocytic leukemia. Cancer Medicine 2021;10:4387-96.
• 22. Falay M, Serdar MA, Dalgali H, Uçar MA, Dagdaş S, Özet G. Which markers should the used for diagnostic chronic lymphocytic leukemia immunophenotyping scoring system by flow cytometry. Clinical Laboratory 2019;65:2049-55.
• 23. Ibrahim F, Al Sabbagh A, Amer A, Soliman DS, Al Sabah H. Composite chronic lymphocytic leukemia/small lymphocytic lymphoma and mantle cell lymphoma; small cell variant: a real diagnostic challenge. Case presentation and review of literature. American Journal of Case Reports 2020;21:e921131-1.
• 24. Moreau EJ, Matutes E, A’Hern RP, et al. Improvement of the chronic lymphocytic leukemia scoring system with the monoclonal antibody SN8 (CD79b). Am J Clin Pathol 1997;108:378-82.