Flow Cytometric Analysis of Benign and Malignant Tumors

流式细胞术免疫分型是一种灵敏的良恶性肿瘤分析技术。我们正在研究该技术的改进及其在不同系统中诊断和测量预后标记物的应用。原发性积液性淋巴瘤（PEL）是一种血液淋巴样肿瘤，表现为浆液性积液，无法检测到肿瘤肿块。淋巴瘤细胞表达的标志物很少，有时诊断困难，治疗后疾病的定量也有问题。我们已经开发了一种流式细胞测定法，可用于诊断这种淋巴瘤和定量患者标本中的肿瘤细胞。家族性CLL家族正在被研究，因为它们可能为研究CLL（一种常见的肿瘤）的发展提供了独特的机会。我们研究了普通CLL和家族性CLL患者的免疫表型，发现两组之间存在显著差异。这些数据与生存率的相关性正在进行中。我们已经开发了方法来定量抗体结合能力的肿瘤细胞存在于低水平的淋巴结，外周血或骨髓的患者接受抗体为基础的治疗。这种流式细胞分析代替了以前使用放射性标记抗体来测量患者肿瘤细胞结合能力的研究。流式细胞分析更快速，不需要使用放射性，并且在100微升血液中进行，而不是25mL。此外，该数据更具特异性，因为抗体结合水平是在肿瘤细胞中特异性测量的，而不受患者标本中存在的其他群体的影响。收集到的数据也更加精确，并将用于比较抗体与肿瘤细胞的结合和治疗反应。最近使用流式细胞术的研究表明，在癌症患者的循环中可以检测到内皮细胞前体和成熟内皮细胞，并且这些细胞在恶性肿瘤患者中的水平明显高于正常对照。此外，对恶性过程的成功治疗可能会影响外周血中活化的循环内皮细胞（CEC）的数量。这些数据表明，癌症患者外周血中CEC和循环内皮细胞前体（CEP）的测量可能被用作抗癌策略有效性的替代标志物，特别是那些针对肿瘤血管系统的策略。我们已经开发了一种方法来测量癌症患者或接受手术干预的患者外周血中CECs和cep的水平，并确定这些测量是否可以用于跟踪各种组织学和治疗的反应或疾病进展。

Flow Cytometric immunophenotyping is a sensitive technique for analysis of benign and malignant tumors. We are studying the refinement of this technique and its application to diagnosis and measurement of prognostic markers in different systems. Primary effusion lymphoma (PEL) is a hematolymphoid neoplasm that presents in serous effusions without detectable tumor masses. The lymphoma cells express few markers making diagnosis difficult at times and quantitation of disease following therapy problematic. We have developed a flow cytometric assay that is useful in diagnosis of this lymphoma and in quantitating tumor cells in patient specimens. Familial CLL families are being studied because they potentially provide a unique opportunity to study the development of CLL, a common neoplasm. We have studied the immunophenotypic profiles of patients with common CLL and with familial CLL and found significant differences between the two groups. Correlation of this data with survival is ongoing. We have developed methods to quantitate antibody binding capacity in tumor cells present in low levels in lymph node, peripheral blood or bone marrow for patients under going antibody based therapy. This flow cytometric assay is performed in place of previous studies using radioactively labeled antibodies to measure tumor cell binding capacity in patients. The flow cytometric assay is more rapid, makes the use of radioactivity unnecessary and is performed on 100 micro liters of blood instead of 25mL. Furthermore, the data is more specific as levels of antibody binding are measured specifically in the tumor cells without the influence of other populations present in patient specimens. The data collected is also more precise and will be used to compare antibody binding to tumor cells to treatment response. Recent studies, using flow cytometry have demonstrated that endothelial cell precursors as well as mature endothelial cells can be detected in the circulation of patients with cancer and that the levels of these cells are significantly higher in patients with malignancy than normal controls. Furthermore, successful therapy for the malignant process may affect the number of activated circulating endothelial cells (CEC's) in the peripheral blood. These data suggest that measurements of CEC's and circulating endothelial cell precursors (CEP's) in the peripheral blood of cancer patients may be a used as a surrogate marker for the efficacy of anti-cancer strategies, particularly those that target tumor vasculature. We have developed methodology to measure levels of CECs and CEPs in the peripheral blood of patients with cancer or of patients undergoing surgical interventions, and to determine if these measurements can be used to follow response or disease progression across a variety of histologies and treatments.