AGE ASSOCIATED CHANGES IN B LYMPHOCYTE FUNCTION

本研究计划的长远目标是了解 与年龄相关的免疫异常的潜在基础。 的 具体目标是确定定性和定量变化 在老年动物的B淋巴细胞中。 这些研究将集中在年龄 B细胞中相关的本征缺陷和B细胞中的变化 异质性 将通过以下方法分析B淋巴细胞亚群： 免疫荧光技术结合荧光激活的 细胞分选仪和抗B细胞表面分子的单克隆抗体。 功能测定将包括分析B细胞增殖和 - 响应于各种抗受体抗体的分化， 淋巴因子 这一假说认为，增加自身抗体的生产， 老年动物是一种独特的B异常表现的结果 将检测淋巴细胞亚群。 七种VH基因探针已被证明可以识别所有的鼠源性 将使用重链可变区基因家族。 表达 这些基因将通过分析来自有丝分裂原的标记RNA来确定 年轻和年老动物的活化B细胞。 的相对表示 每个VH基因家族也将在纯化的抗原中确定 特异性B细胞。 剧目中的任何差异将由 杂交瘤的构建和Southern杂交分析VH基因的使用 印迹杂交技术。 这些研究将有助于确定年龄相关的变化 这是B细胞群体固有的，可能有助于设计策略， 有利地调节对老年个体的免疫应答。

The long-term objective of this research proposal is to understand the underlying basis for the immune abnormalities associated with age. The specific Aims are to determine qualitative as well as quantitative changes in the B lymphocytes from aged animals. The studies will focus on the age associated intrinsic defects in B cells and on the changes in the B cell heterogeneity. Subpopulations of B lymphocytes will be analysed by immunofluorescence techniques in conjunction with a fluorescence activated cell sorter and monoclonal antibodies against B cell surface molecules. Functional assays will include analysis of the B cell proliferation and differentiation in response to various anti-receptor antibodies and lymphokines. The hypothesis that the increased auto-antibody production in aged animals is the regult of abnormal representation of a unique B lymphocyte subpopulation will be tested. Seven VH gene probes that have been shown to identify all of the murine heavy chain variable region gene families will be employed. Expression of these genes will be determined by analysis of labelled RNA from mitogen activated B cells from young and old animals. Relative representation of each of the VH gene families will also be determined in purified antigen specific B cells. Any differences in the repertoire will be confirmed by construction of hybridomas and analysis of the VH gene useage by Southern blot hybridization techniques. These studies will be instrumental in defining the age associated changes that are intrinsic to B cell populations and may help devise strategies to favorably modulate the immune responses to aged individuals.