Multiparameter Flow Cytometry to Detect Hematogones and to Assess B-lymphocyte clonality in Bone Marrow Samples from Patients with Non-Hodgkin Lymphomas

多参数流式细胞术检测非霍奇金淋巴瘤患者骨髓样本中的造血细胞并评估 B 淋巴细胞克隆性

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作者:Giovanni Carulli, Virginia Ottaviano, Valentina Guerri, Stefano Giuntini, Paola Sammuri, Eugenio M Ciancia, Antonio Azzarà

Abstract

Hematogones are precursors of B-lymphocytes detected in small numbers in the bone marrow. Flow cytometry is the most useful tool to identify hematogones and, so far, 4-color methods have been published. In addition, flow cytometry is used in the diagnosis and follow-up of lymphomas. We developed a flow cytometric 7-color method to enumerate hematogones and to assess B-lymphocyte clonality for routine purposes. We evaluated 171 cases of B-cell non-Hodgkin lymphomas, either at diagnosis or in the course of follow-up. By our diagnostic method, which was carried out by the combination K/λ/CD20/CD19/CD10/CD45/CD5, we were able to detect hematogones in 97.6% of samples and to distinguish normal B-lymphocytes, neoplastic lymphocytes and hematogones in a single step. The percentage of hematogones showed a significant inverse correlation with the degree of neoplastic infiltration and, when bone marrow samples not involved by disease were taken into consideration, resulted higher in patients during follow-up than in patients evaluated at diagnosis.

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