Abnormalities by Multicolor Flow Cytometry for Detection of Minimal Residual Disease in Recipients of Allo-HSCT Originating from Donors: A Cohort StudyHui Wang1, Aixian Wang1, Man Chen2, Meiwei Gong1, Xueying Wu1, Junyi Zhen1, Yue Lu31Hebei Yanda Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Langfang, China
2Beijing Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Beijing, China
3Hebei Yanda Lu Daopei Hospital, Department of Stem Cell Transplantation, Langfang, China
Objective: In minimal residual disease (MRD) analysis after allogeneic hematopoietic stem cell transplantation (allo-HSCT), abnormal immunophenotyping is commonly considered as evidence of a secondary recurrence or complications, leading to overtreatment. We aimed to confirm whether such phenotypic abnormality might originate from donors using multicolor flow cytometry (MFC).
Materials and Methods: The MRD of bone marrow specimens of 3395 patients who had received allo-HSCT were analyzed using the conventional two-tube, eight-color MFC panel. The frequencies of abnormal immunophenotypes were also evaluated in three groups of patients without malignancies.
Results: The frequency of new abnormal polymorphisms was 0.088% (3/3395) among patients who received allo-HSCT. The abnormal cells seen in three patients in complete remission were Fcγ receptor IIIB (FcγRIIIB) gene deletion (CD16- neutrophils), CD2-CD159a-CD159c+ natural killer (NK) cells, and monoclonal B lymphocytosis (MBL), respectively. In addition, abnormal T-cells (CD4+CD8+) were detected in one donor before allo-HSCT. Identical abnormalities were found in the peripheral blood of the corresponding donors of the three patients via MFC. Among the individuals without malignancies, the incidence of FcγRIIIB deletion was 0.2% (11/5256), that of NK cells with the absence of CD2 and single-positive CD159c was 0.05% (1/2000), that of monoclonal CD4/CD8 double-positive T-cells was 0.05% (1/2000), and that of MBL was 1.3% (14/1100). The frequency of NK cells with the absence of CD2 was 1.3% (1/79) and with CD8dim was 14% (11/79) in NK cell lymphoma. The following abnormalities could be identified by the two-tube, eight-color MFC panel: cκ/cλ/CD19/CD5/CD20/ CD38/CD45/CD56 (adding CD10 and CD34 as the ninth and tenth colors) and CD16+CD56/CD5/CD3/CD7/CD4/CD8/CD2/CD45 (adding CD117 as the ninth color).
Conclusion: Abnormalities in recipients of allo-HSCT detected by MRD analysis may originate from their donors. Screening of donor specimens with a suitable two-tube, eight- to ten-color MFC panel may be a promising method for minimizing misdiagnoses.
Keywords: Multicolor flow cytometry, Allogeneic hematopoietic stem cell transplantation, Fcγ, RIIIB deletion, Monoclonal B lymphocytosis, Monoclonal CD4+CD8+ T-cells, CD2-CD159c+ natural killer cells
Allo-KHN Alıcılarında Minimal Kalıntı Hastalık Tespitindeki çok Renkli Akım Sitometri ile Saptanan Verici Kaynaklı Anormallikler: Bir Kohort ÇalışmasıHui Wang1, Aixian Wang1, Man Chen2, Meiwei Gong1, Xueying Wu1, Junyi Zhen1, Yue Lu31Hebei Yanda Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Langfang, China
2Beijing Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Beijing, China
3Hebei Yanda Lu Daopei Hospital, Department of Stem Cell Transplantation, Langfang, China
Amaç: Allojeneik hematopoetik kök hücre nakli (allo-KHN) sonrası minimal kalıntı hastalık (MKH) analizinde anormal immünofenotiplendirme sıklıkla sekonder rekürrensin ya da komplikasyonların kanıtıdır, neticede fazla tedaviye yol açar. Bu tarz fenotipik anormalliklerin çok renkli akım sitometri (ASM) kullanarak verici kökenli olabileceğini göstermek ve bunu doğrulamayı hedefledik.
Gereç ve Yöntem: Allo-KHN olmuş 3395 hastanın kemik iliği örneklerinde konvansiyonel iki tüp sekiz renkli ASM paneli ile MKH bakıldı. Anormal immünfenotiplenmenin sıklıkları da malignitesi olmayan üç grup hastada değerlendirildi.
Bulgular: Allo-KHN olan hastalarda yeni anormal polimorfizmlerin sıklığı %0,088 (3/3395) idi. Tam remisyondaki üç hastada mevcut anormal hücreler sırasıyla Fcγ reseptörü IIIB (FcγRIIIB) gen delesyonu (CD16 nötrofil), CD2CD159aCD159c doğal öldürücü (NK) hücreleri ve monoclonal B lenfositoz (MBL) idi. Ilaveten, bir vericide allo-KHN öncesinde anormal T-hücreleri (CD4CD8) tespit edilmişti. Benzer anormallikler üç hastanın vericilerinin periferik kanlarında ASM ile bulunmuştur. Malignitesi olmayan bireylerde FcγRIIIB delesyonu insidansı %0,2 (11/5256), CD2 si olmayan NK hücreleri ve tek CD159c pozitif olanlar %0,05 (1/2000), monoclonal CD4/CD8 çift pozitif T-hücreleri %0,05 (1/2000), MBL %1,3 (14/1100) bulundu. NK hücreli lenfomada CD2 negatif NK hücreleri sıklığı %1,3 (1/79) ve CD8 dim olanlar %14 (11/79) idi. Sıradaki anormallikler çift tüp sekiz renkli ASM paneli ile tanımlanmıştır: cκ/cλ/CD19/CD5/CD20/CD38/CD45/ CD56 (CD10 ve CD34 dokuzuncu ve onuncu renk olarak eklenmiştir) ve CD16+CD56/CD5/CD3/CD7/CD4/CD8/CD2/CD45(CD117 dokuzuncu renk olarak eklenmiştir).
Sonuç: Allo-KHN alıcılarında MKH ile tespit edilen anormallikler vericilerinden kaynaklanabilir. Verici örneklerinin çift tüp sekiz-on renkli ASM paneli ile taranması yanlış tanıları en aza indirmek için ümit veren bir yöntemdir.
Anahtar Kelimeler: Çok renkli akım sitometri, Allojeneik hematopoetik kök hücre nakli, Fcγ, RIIIB delesyonu, Monoklonal B lenfositoz, Monoklonal CD4+CD8+ T-hücreleri, CD2-CD159c+ Doğal öldürücü hücreler
Hui Wang, Aixian Wang, Man Chen, Meiwei Gong, Xueying Wu, Junyi Zhen, Yue Lu. Abnormalities by Multicolor Flow Cytometry for Detection of Minimal Residual Disease in Recipients of Allo-HSCT Originating from Donors: A Cohort Study. Turk J Hematol. 2023; 40(1): 18-27
Corresponding Author: Hui Wang, China |
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