CELLULAR AND MOLECULAR INTERACTIONS IN AGING HUMANS

总的目标是了解细胞和分子基础。 老年人类免疫调节异常与唐氏综合征(DS)这个 自体混合淋巴细胞反应(AMLR)和B细胞活化 以增殖和分化为模型。在AMLR中， 不同的T细胞亚群增殖、产生和应答可溶性T细胞 调节因子，表达不同的免疫调节功能。休息B 细胞对B细胞生长因子(BCGF)的作用具有抵抗力；然而， 激活后通过增殖反应BCGF，因此，直接 B细胞对BCGF的反应提示B细胞在体内被激活。 具体目的是：(A)研究增殖和免疫调节 T-非T和T-TA激活的T细胞及其亚群的功能 (B)研究白细胞介素1(IL-1)的产生和影响； IL-2和BCGF；(C)研究细胞的增殖和分化 单抗定义的B细胞及其亚群；(D)计数 T细胞和B细胞亚群的数量和比例，使用 单抗；和(E)将定量和/或 免疫调节性T细胞的质异常和T细胞的异常 有或无自身抗体的B细胞。大致 老年人(65岁以上)和年轻人(18-35岁)各80-90人，40人 将对15-35岁的DS患者进行研究。T细胞和非T细胞 由E-ROSETING分隔。T细胞和B细胞亚群均为阳性 使用单抗(OKT4、OKT8和FMC1、FMC7)进行阴性选择 和平底锅技术。T-Non T AMLR中激活的T细胞/T细胞亚群 连续4天(TA)照射，作为T-TA AMLR的刺激物。T 治疗T-Non T和T-TA AMLR中激活的细胞/T细胞亚群 用丝裂霉素C检测T细胞的辅助/抑制功能 AMLR细胞增殖与免疫球蛋白B细胞分化的关系 合成和分泌细胞。人外周血中产生的IL-2活性 PHA或ConA激活的T细胞产生AMLR、BCGF和IL-1 用脂多糖刺激的巨噬细胞在CTLL-6细胞系上进行了研究 抗Mu激活的B细胞和IL-1依赖细胞系的增殖 分别进行了分析。用电池研究T和B淋巴细胞亚群 用单抗和流式细胞仪检测。这些观察结果将有所帮助 在了解衰老和DS的免疫失调机制方面 有助于设计可在以下方面使用的方法 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.