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 调节造血系统的发育，正常以及 恶性的 已经认识到IL-6在免疫调节中具有主要作用。 多发性骨髓瘤的发展，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- 6刺激，使我们相信IL-6基因表达， 骨髓瘤细胞是诱导型的而不是组成型的。 识别 通过骨髓瘤细胞诱导IL-6基因表达的细胞因子， 确定产生它们的细胞是这项研究的目标之一， 项目 将采用PCR和ELISA等敏感技术 和单细胞分析的新技术。 虽然骨髓瘤 细胞产生IL-6并展示IL-6受体，它们在体内的低表达 增殖活性和细胞因子无法诱导 体外持续增殖表明IL-6的靶细胞 可以是骨髓瘤前体细胞。 为了解决这个问题，方法 用于从骨髓瘤患者的血液中纯化前骨髓瘤细胞， 开发 基于表型的高分辨率流式分选 报告的与前骨髓瘤细胞相关的特征将被 用于获得高纯度的这些细胞。 细胞因子对 这些细胞的增殖和分化成 可识别的单克隆骨髓瘤细胞将使用多种 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， Ki-67抗原的表达将用于确定增殖。 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。 审查 地塞米松和干扰素的作用，这是有效的， 骨髓瘤的治疗将揭示它们是否对骨髓瘤细胞有调节作用。 细胞因子的产生，如果对这些药物的耐药性是肿瘤 和/或辅助细胞现象。 这些数据是在帮助下生成的 将阐明细胞因子在发展中的作用， 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.