Flow Cytometric Analysis of Benign and Malignant Tumors

流式细胞术免疫表型分析是一种敏感的技术，用于分析良性和恶性肿瘤。我们正在研究这项技术的改进及其在不同系统中诊断和测量预后标志物的应用。我们已经开发了对正在进行基于抗体的治疗的患者的淋巴结、外周血或骨髓中以低水平存在的肿瘤细胞中的抗体结合能力进行定量的方法。该流式细胞术测定代替先前使用放射性标记抗体测量患者中肿瘤细胞结合能力的研究。流式细胞术分析更快速，无需使用放射性，并且对100 mL血液而不是25 mL血液进行。此外，数据更具特异性，因为在肿瘤细胞中特异性测量抗体结合水平，而不受患者标本中存在的其他群体的影响。收集的数据也更精确，并将用于比较抗体与肿瘤细胞的结合与治疗反应。最近的研究，使用流式细胞术已经证明，内皮细胞前体细胞以及成熟的内皮细胞可以检测到癌症患者的循环中，这些细胞的水平显着高于恶性肿瘤患者比正常对照。此外，恶性过程的成功治疗可能会影响外周血中活化循环内皮细胞（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）的测量值可用作抗癌策略（特别是靶向肿瘤血管系统的那些）功效的替代标志物。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CEC和CEP的水平，并确定这些测量是否可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. 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 mL 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.