Flow cytometric detection of malignant cells in body fluids

原发性积液性淋巴瘤是一种由HIV和HHV8病毒引起的侵袭性淋巴瘤，主要涉及体液。多中心Castlemans病是一种在相同临床环境下发生的非克隆增生过程。这些罕见病的形态学诊断比较困难。我们已经开发了临床流式细胞术分析，可以帮助诊断原发性积液性淋巴瘤和多中心Castlemans病，为NCI方案的患者提供关键的临床支持。造血系统恶性肿瘤累及脑脊液（CSF）也很难单凭形态学来证实。在一项比较形态学与流式细胞术形态学的研究中，我们证明了流式细胞术免疫表型分型在建立一系列已知淋巴瘤或白血病患者的肿瘤诊断以及使用形态学标准对“非典型”或“可疑”脑脊液的初步诊断方面是有用的。流式细胞术小组评估了流式细胞术分析在高级别B细胞淋巴瘤患者的分期和治疗中的作用。流式细胞术鉴定的肿瘤克隆仅占脑脊液淋巴细胞总数的0.02%。流式细胞术检测到细胞学、化学和细胞计数失败的受累部位。流式细胞术检测疾病是一个负面的预后因素，现在提示治疗干预。根据这项研究，建议有侵袭性B细胞淋巴瘤累及中枢神经系统风险的患者通过流式细胞术进行脑脊液分期评估。流式细胞术组参加了一次国际共识会议，以确定脑脊液流式细胞术分析的最佳方法。

Primary effusion lymphoma is an aggressive lymphoma arising in the setting of HIV and HHV8 that primarily involves body fluids. Multicentric Castlemans disease is a non-clonal proliferative process that arises in the same clinical setting. Morphological diagnosis is difficult in these rare diseases. We have developed clinical flow cytometric assays that can aid in the diagnosis of primary effusion lymphoma and multicentric Castlemans disease, providing key clinical support to patients on NCI protocols. Involvement of the cerebrospinal fluid (CSF) by hematopoietic malignancies may also be difficult to document by morphology alone. In a study comparing morphology alone to morphology with flow cytometry, we demonstrated that flow cytometric immunophneotyping was useful in establishing a diagnosis of neoplasia in a series of patients with known lymphoma or leukemia and an initial diagnosis of "atypical" or "suspicious" CSF using morphologic criteria. The Flow Cytometry Unit evaluated the role of flow cytometric analysis in staging and management of patients with high grade B cell lymphomas. Flow cytometry identified neoplastic clones that constituted as little as 0.02% of total CSF lymphocytes. Flow cytometry detected involvement where cytology, chemistry and cell counts failed. Flow cytometric detection of disease was a negative prognostic factor and now prompts therapeutic intervention. As a result of the study it is recommended that patients at risk for CNS involvement by aggressive B cell lymphoma undergo staging CSF evaluation by flow cytometry. The Flow Cytometry Unit participated in an international consensus conference to determine optimal methodology for flow cytometric analysis of CSF.