CELLULAR AND MOLECULAR INTERACTIONS IN AGING HUMANS

总体目标是了解细胞和分子基础， 老年人的免疫调节异常和唐氏综合症（DS）。 的 自体混合淋巴细胞反应（AMLR）和B细胞活化， 增殖和分化用作模型。 在AMLR中， 不同的T细胞亚群增殖，产生和应答可溶性 介质，并表达不同的免疫调节功能。 静止B 细胞对B细胞生长因子（BCGF）的作用有抗性;然而， 活化后通过增殖对BCGF作出反应，因此， B细胞对BCGF的应答表明B细胞的体内活化。 具体目的是：（a）研究增殖和免疫调节 T细胞和T细胞亚群在T-非T和T-TA中活化的功能 AMLR;（B）研究白细胞介素-1（IL-1）的产生和影响， IL-2和BCGF;（c）研究人乳腺癌细胞株的增殖和分化。 单克隆抗体定义的B细胞和B细胞亚群;（d）计数 T细胞和B细胞亚群的数量和比例， 单克隆抗体;和（e）将定量和/或 免疫调节性T细胞的定性异常和 存在或不存在自身抗体的B细胞。 约 老年（大于65岁）和年轻（18 - 35岁）受试者各80 - 90例， 将对DS患者（15 - 35岁）进行研究。 T细胞和非T细胞 以E玫瑰花分隔。 T细胞和B细胞亚群呈阳性， 使用单克隆抗体（OKT 4、OKT 8和FMC1、FMC7）进行阴性选择 和平移技术。 T细胞/T细胞亚群在T-非T AMLR中活化 4天（TA），并用作T-TA AMLR中的刺激物。 不 治疗在T-非T和T-TA AMLR中活化的T细胞/T细胞亚群 丝裂霉素C和检查辅助/抑制功能， AMLR中的细胞增殖和IG的B细胞分化 合成和分泌细胞。 IL-2的活性在细胞内表达， 由PHA或Con A激活的T细胞产生的AMLR、BCGF和产生的IL-1 在CTLL-6细胞系上研究LPS刺激的巨噬细胞， 抗Mu激活的B细胞和IL-1依赖性细胞系的增殖 分别 T淋巴细胞和B淋巴细胞亚群的研究使用电池 单克隆抗体和流式细胞仪。 这些观察将有助于 了解衰老和DS中免疫失调的机制， 可有助于设计可用于 延缓与衰老相关的现象和疾病。

The overall objective is to understand the cellular and molecular bases of abnormal immune regulation in aging humans and Down's syndrome (DS). The autologous mixed lymphocyte reaction (AMLR) and B cell activation, proliferation and differentiation are used as models. In the AMLR, distinct subsets of T cells proliferate, produce and respond to soluble mediators, and express distinct immunoregulatory functions. Resting B cells are resistant to the effect of B cell growth factor (BCGF); however, after activation respond to BCGF by proliferation, therefore, direct response of B cells to BCGF would suggest in vivo activation of B cells. The specific aims are: (a) to study the proliferative and immunoregulatory functions of T cells and T cells subsets activated in T-non T and T-TA AMLR; (b) to study the production and influence of interleukin-1 (IL-1), IL-2 and BCGF; (c) to study the proliferation and differentiation of monoclonal antibody-defined B cells and B cell subsets; (d) to enumerate the numbers and proportions of T cell- and- B cell subsets, using monoclonal antibodies; and (e) to correlate the quantitative and/or qualitative abnormalities of immunoregulatory T cells and abnormalities of B cells with the presence or absence of autoantibodies. Approximately 80-90 each aging (greater then 65 years) and young (18-35) subjects and 40 patients (15-35 years) with DS will be studied. T and non T cells are separated by E-rosetting. T cell- and B cell subsets are positively and negatively selected using monoclonal antibodies (OKT4, OKT8 and FMC1, FMC7) and panning technique. T cells/T cell subsets activated in T-non T AMLR for 4 days (TA) are irradiated and used as stimulators in T-TA AMLR. T cells/T cell subsets activated in the T-non T and T-TA AMLR are treated with mitomycin C and examined for helper/suppressor functions against T cell proliferation in the AMLR and B cell differentiation for Ig synthesizing and secreting cells. The activity of IL-2 produced in the AMLR, BCGF produced by T cells activated by PHA or Con A and IL-1 produced by LPS-stimulated macrophages are studied on CTLL-6 cell line, on proliferation of anti-Mu activated B cells, and IL-1 dependent cell line respectively. Subsets of T- and B- lymphocytes are studied using a battery of monoclonal antibodies and FACS analyzer. These observations will help in understanding the mechanisms of immunodysregulation in aging and DS that could help in designing the approach(es) that could be utilized in postponing the phenomena and diseases associated with aging.