FLOW CYTOMETRIC ANALYSIS OF DNA IN HUMAN LYMPHOMA CELLS

人淋巴瘤细胞 DNA 的流式细胞术分析

• 批准号: 3177653
• 负责人: RAUL C. BRAYLAN
• 金额: $ 11.15万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1988-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3177653
• 关键词: B lymphocyte; Epstein Barr virus; T lymphocyte; aneuploidy; biopsy; bromodeoxyuridine; cytogenetics; flow cytometry; fluorescent dye /probe; human tissue; immune adherence reaction; immunoglobulins; leukocyte activation /transformation; lymphoma; monoclonal antibody; neoplasm /cancer genetics; neoplasm /cancer relapse /recurrence; neoplastic cell; neoplastic growth; radiotracer; tumor antigens

Flow cytometry (CFM) allows the rapid measurement of antigens and nucleic acids in individual cells. DNA analysis by FCM is particularly useful because it provides rapid information on cell cycle kinetics and tumor ploidy. B-cell lymphomas are kinetically heterogeneous tumors composed of immunoglobulin-bearing lymphocytes. Curiously, these tumor often contain large numbers of normal T-cells which hamper the precise analysis of DNA in tumor B-cells. By labelling cell surface immunoglobulins, we measured DNA in the neoplastic cells of B-cell lymphomas without interference from T-cells. The DNA synthetic fraction for the neoplastic cells correlated remarkabley well with histologic type and it should be an useful indicator of tumor proliferative activity. By comparing the DNA content of tumor B-cells with that of non-neoplastic T-cells from the same tissue, we detected minimal (2%) aneuploidy in neoplastic cells. We also observed two neoplastic populations with different DNA content in approximately 10% of the tumors suggesting ciclonality. In the present study we propose to answer the following: What is the nature of aneuploidy detected by FCM in the B-cell lymphomas? Does it only reflect genomic size or is it in part due to binding properties of the fluorescent dyes to neoplastic or cycling cells? Is the conversion of low grade lymphomas into high grade lymphomas accompanied by ploid or kinetic changes? What is the nature of the "biclonality" observed in 10% of B-cell lymphomas? Since most of the second "clones" in these cases are near-tetraploid, do they represent arrested or cycling cells? If they are cycling, is the DNA synthetic rate of the aneuploid cells higher or lower than that of the diploid cells of the same tumor? Are multiple "clones" related to previous therapy? Is there a relationship between T-cell subset distribution within the lymph node and the kinetic properties of the neoplastic B-cells? We plan to answer these questions as follows: Modal chromosome number determined on metaphases obtained in direct preparations of tumor cells or in normal and tumor cells induced by stimulation with B-cell mitogens will be compared with the DNA index obtained by FCM. Various DNA fluorescent probes will be tested to determine if ploidy changes reflect DNA binding properties of the fluorescent dyes. DNA indices and kinetics properties of recurrent tumors will be compared with those obtained in previous biopsies. Simultaneous analysis of cellular DNA content and bromodeoxyuridine incorporation will be performed to establish if aneuploid cells are cycling and their rate of DNA synthesis. Correlations will be made between therapy prior to biopsy and tumor ploidy. T-lymphocyte subsets within the tumors will be analyzed to assess if a relationship exists between T-cell distribution and kinetic properties of the neoplastic B-cells.

流式细胞术（CFM）允许快速测量抗原和核酸。 酸在单个细胞中。 FCM的DNA分析特别有用 因为它提供了关于细胞周期动力学和肿瘤的快速信息， 倍性 B细胞淋巴瘤是一种动力学异质性肿瘤， 带有免疫球蛋白的淋巴细胞。 奇怪的是，这些肿瘤通常含有 大量正常T细胞阻碍了DNA的精确分析 肿瘤B细胞 通过标记细胞表面免疫球蛋白，我们测量了DNA 在B细胞淋巴瘤的肿瘤细胞中， T细胞 肿瘤细胞的DNA合成分数相关 与组织学类型相关性良好，应作为一个有用的指标 肿瘤增殖活性。 通过比较肿瘤DNA含量 B细胞与非肿瘤性T细胞来自同一组织，我们 在肿瘤细胞中检测到最小（2%）的非整倍体。 我们还观察到两个 在大约10%的肿瘤人群中具有不同DNA含量， 肿瘤显示有环状病变 在本研究中，我们建议 回答以下问题：FCM检测到的非整倍体的性质是什么？ B细胞淋巴瘤 它仅仅反映了基因组的大小，还是 由于荧光染料对肿瘤或循环的结合特性 细胞？ 低级别淋巴瘤转化为高级别淋巴瘤 伴随着倍体或动力学变化？ 的本质是什么 在10%的B细胞淋巴瘤中观察到“双克隆性”？ 由于大部分 第二个“克隆”在这些情况下是近四倍体，他们代表 被捕还是在牢房里转 如果它们在循环， 非整倍体细胞比二倍体细胞高或低， 同样的肿瘤？ 多次“克隆”是否与既往治疗有关？ 是 淋巴内T细胞亚群的分布 节点和肿瘤B细胞的动力学特性？ 我们计划 回答这些问题如下：模态染色体数目确定 在肿瘤细胞的直接制备物中或在正常和 将比较用B细胞有丝分裂原刺激诱导的肿瘤细胞 用流式细胞仪测定DNA指数。 各种DNA荧光探针将被 测试以确定倍性变化是否反映了细胞的DNA结合特性。 荧光染料。 复发肿瘤的DNA指数和动力学特征 将与先前活检中获得的结果进行比较。 同时 将分析细胞DNA含量和溴脱氧尿苷掺入。 以确定非整倍体细胞是否在循环，以及它们的DNA 合成. 将在活检前的治疗与 肿瘤倍性 将分析肿瘤内的T淋巴细胞亚群， 评估T细胞分布和动力学之间是否存在关系 肿瘤性B细胞的特性。