Flow Cytometric Analysis of Benign and Malignant Tumors

流式细胞免疫分型技术是分析良、恶性肿瘤的一种灵敏技术。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记物的应用。原发性渗出性淋巴瘤(PEL)是一种血液淋巴样肿瘤，表现为浆液性积液，未发现肿块。淋巴瘤细胞表达的标记物很少，这使得诊断有时很困难，治疗后疾病的量化也存在问题。我们已经开发了一种流式细胞术分析，这对诊断这种淋巴瘤和对患者标本中的肿瘤细胞进行定量是有用的。家族性CLL家族正在被研究，因为他们可能为研究CLL这一常见肿瘤的发展提供了一个独特的机会。我们研究了普通CLL患者和家族性CLL患者的免疫表型特征，发现两组患者之间存在显著差异。这些数据与存活率的关联正在进行中。我们已经为接受抗体治疗的患者开发了定量检测淋巴结、外周血或骨髓中低水平肿瘤细胞抗体结合能力的方法。这种流式细胞术分析取代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。流式细胞术分析更快速，不需要使用放射性，可以对100微升的血液进行检测，而不是25毫升。此外，由于抗体结合水平是在肿瘤细胞中专门测量的，而不受患者样本中存在的其他群体的影响，因此数据更具特异性。收集的数据也更准确，将用于比较与肿瘤细胞结合的抗体与治疗反应。最近利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，恶性过程的成功治疗可能会影响外周血中激活的循环内皮细胞(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记物，特别是那些针对肿瘤血管的策略。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方法的反应或疾病进展。

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.