Flow Cytometry Unit, Pathology Division, Citta' della Salute e della Scienza Hospital, Turin, Italy.
Pathology Division, Citta' della Salute e della Scienza Hospital, Turin, Italy.
Int J Lab Hematol. 2019 Apr;41(2):208-217. doi: 10.1111/ijlh.12946. Epub 2018 Nov 27.
Nonhemopoietic neoplasms (NHNs) may be encountered during routine flow cytometry (FC) immunophenotyping. The clue of their presence mainly relies on detection of CD45-negative (CD45-) cells with altered scatter parameters.
In this study, we evaluated a monoclonal antibody combination conceived to characterize the CD45- population by FC, suspected of belonging to NHNs, when present. The panel included CD45 for leucocytes identification, CD326 (clones BerEP4 and HEA-125) to mark epithelial cells, CD33 to identify myeloid cells, CD138 to trace plasma cells and CD56 useful in the identification of neuroendocrine tumours. 7AAD vital dye was used to gate out dead cells. Results were correlated with cytomorphology and confirmed by histological data, if available.
Among 9422 specimens submitted for routine FC investigation, 47 samples that included fine-needle aspirates, bone marrow aspirates, tissue biopsies and body fluids had a detectable CD45- population and a sufficient cell amount to be further investigated. FC revealed the presence of CD326-positive epithelial cells in 38 specimens; altered scatter parameters and variable reactivity to the other antigens tested allowed to suspect NHNs in the remaining nine samples. The presence of NHNs was confirmed in all cases by morphology.
The current results show that when CD45- cells with altered scatter parameters were detected, cytometrists involved in leukaemia/lymphoma diagnosis may require further FC investigations to rapidly identify NHNs in different specimens, thus reducing the time of the immunohistochemical diagnostic workup to reach a final diagnosis.
非造血性肿瘤（NHN）在常规流式细胞术（FC）免疫表型分析中可能会遇到。其存在的线索主要依赖于检测 CD45 阴性（CD45-）细胞的散射参数改变。
在这项研究中，我们评估了一组设计用于通过 FC 来对 CD45- 群体进行特征分析的单克隆抗体组合，当存在疑似 NHN 时，这些抗体可以被检测到。该组合包括用于白细胞识别的 CD45、标记上皮细胞的 CD326（BerEP4 和 HEA-125 克隆）、用于识别髓系细胞的 CD33、用于追踪浆细胞的 CD138 和有助于神经内分泌肿瘤识别的 CD56。7AAD 活染料用于排除死亡细胞。结果与细胞形态学相关联，并在有组织学数据的情况下进行确认。
在 9422 份提交进行常规 FC 检查的标本中，47 份包括细针抽吸物、骨髓抽吸物、组织活检和体液的标本中存在可检测的 CD45- 群体，且细胞数量足以进一步进行研究。FC 在 38 份标本中发现了 CD326 阳性上皮细胞；改变的散射参数和对其他测试抗原的可变反应性使得在其余 9 份标本中怀疑存在 NHN。在所有情况下，形态学都证实了 NHN 的存在。
目前的结果表明，当检测到 CD45- 细胞散射参数改变时，参与白血病/淋巴瘤诊断的细胞仪技术人员可能需要进一步的 FC 研究，以便在不同标本中快速识别 NHN，从而减少免疫组织化学诊断工作的时间，以达到最终诊断。
