CYTOKINE NETWORK IN MYELOMATOSIS--CLUES TO DISEASE MANIFESTATION AND THERAPY

细胞因子、细胞因子受体和基质相互作用控制和 调节正常和正常的造血系统的发育 恶性的。白介素6已被认为在 多发性骨髓瘤的发生，尽管其作用方式尚不清楚。 初步研究表明自分泌和旁分泌IL-2并存。 6刺激并使我们相信IL-6基因的表达通过 骨髓瘤细胞是可诱导的，而不是结构性的。识别 诱导骨髓瘤细胞IL-6基因表达的细胞因子和 确定产生它们的细胞是这项研究的目标之一 项目。将采用聚合酶链式反应和酶联免疫吸附试验等敏感技术。 以及为单细胞分析开发的新技术。虽然骨髓瘤 细胞产生IL-6并显示IL-6受体，其在体内的低水平 增殖活性与细胞因子不能诱导 体外持续增殖提示IL-6的靶细胞 可能是骨髓瘤前体细胞。为了解决这一假设，方法 为了从骨髓瘤患者的血液中提纯骨髓瘤前细胞 发展起来的。基于表型的高分辨率流分选 据报道，与前骨髓瘤细胞相关的特征将是 用来获得高纯度的这些细胞。细胞因子对血管内皮细胞生长的影响 这些细胞的增殖和分化为 可识别的单克隆性骨髓瘤细胞将使用各种 培养条件，包括与细胞因子分泌细胞共培养。 先进的细胞周期分析技术，如BUDR/IUdR标记和 Ki-67抗原的表达将被用来确定增殖情况。 借助图像进行形态、遗传和表型分析 分析将有助于确定差异化。考试的结果 地塞米松和干扰素对慢性粒细胞白血病的治疗作用 骨髓瘤的治疗将揭示它们是否对 细胞因子的产生以及对这些药物的耐药性是否为肿瘤 和/或副细胞现象。这些数据是通过帮助生成的 从核心，将阐明细胞因子在发育中的作用 多发性骨髓瘤。

Cytokines, cytokine receptors and matrix interactions control and regulate the development of the hematopoietic system, normal as well as malignant. IL-6 has been recognized to have a major role in the development of multiple myeloma, although its mode of action is unclear. Preliminary studies indicate co-existence of autocrine and paracrine IL- 6 stimulation and lead us to believe that IL-6 gene expression by myeloma cells is inducible rather than constitutive. Identifying the cytokines which induce IL-6 gene expression by myeloma cells and determining the cells which produce them is one of the goals of this project. Sensitive techniques such as PCR and ELISA will be employed and new techniques developed for single cell analysis. Although myeloma cells produce IL-6 and display IL-6 receptors, their low in vivo proliferative activity and the inability of the cytokine to induce sustained proliferation in vitro suggest that the target cell for IL-6 may be a myeloma precursor cell. To address this hypothesis, methods for purifying the pre-myeloma cells from myeloma patient's blood will be developed. High resolution flow sorting on the basis of phenotypic characteristics reported to be associated with pre-myeloma cells will be used to obtain these cells in high purity. The effects of cytokines on the proliferation and differentiation of these cells into the recognizable monoclonal myeloma cells will be studied using a variety of culture conditions, including co-cultures with cytokine secreting cells. Sophisticated cell cycle analysis techniques like BUdR/IUdR labeling and expression of the Ki-67 antigen will be used to determine proliferation. Morphologic, genetic and phenotypic analysis with the aid of image analysis will help determine differentiation. Examination of the effects of dexamethasone and interferon which are effective in the treatment of myeloma will reveal if they have regulatory effects on the production of cytokines and if resistance to these drugs is a tumor and/or an accessory cell phenomenon. These data, generated with help from the Cores, will elucidate the role of cytokines in the development of multiple myeloma.