NUTRITIONAL COPPER DEFICIENCY AND THE IMMUNE RESPONSE

这项研究的长期目标是阐明生物化学 铜在免疫功能中的作用及其在发育过程中的决定作用 铜对免疫系统最重要。精选生化和 将使用免疫学技术。为了实现这些目标并实现 检验某些假设时，将采用动物模型。我们会引诱 C57BL和C58围产期铜缺乏的研究 从怀孕到成年早期的几个不同时期的小鼠。 将进行再利用研究。 在耗尽期间和之后的再耗尽期间的铜状态将通过 测定红细胞压积和血浆铜蓝蛋白活性。器官 (脾和胸腺)和血清总铜和铁的水平 用原子吸收光谱仪(AAS)测定。功能上的变化 将通过分析铜锌的活动性来评价铜矿藏。 超氧化物歧化酶或细胞色素c氧化酶。 免疫学评估将包括调查体液免疫 检测脾组织中抗体产生细胞的数量 绵羊红细胞挑战赛。细胞免疫的研究将通过 定量测定培养的胸腺细胞和脾细胞的丝裂原反应性 混合淋巴细胞反应。来自脾的淋巴细胞亚群 通过使用单抗的免疫荧光程序来确定 到表面抗原。植物凝集素(PNA和SBA)将用于评估 胸腺、脾和骨中淋巴细胞的成熟发育 骨髓。用酶联免疫吸附试验检测血清中的IgM和Ig G。 将进行额外的生化测量，以阐明 检验特定假说时免疫力受损的机制。 脾和胸腺的淋巴细胞将被分离出来，血浆 膜的特征是脂肪和蛋白质组成的变化。 分别采用气相色谱和凝胶电泳法。 淋巴器官的抗氧化状态将通过量化来评估 特定的代谢物。将对铜矿资源进行回收研究 或葡萄糖酸亚铁和纯化的小鼠铜蓝蛋白或转铁蛋白 确定免疫力改变的程度，如果有的话，是铁依赖的。 血浆、脑和脾的去甲肾上腺素水平将通过 高效液相色谱-电化学检测。铜的代谢将在#年进行研究。 ~(67)Cu和原子吸收光谱显示淋巴器官的发育。

The long-range objective of this research is to elucidate the biochemical roles of copper in immune function and to determine when during development copper is most critical to the immune system. Selected biochemical and immunological techniques will be used. To accomplish these goals and to test certain hypotheses, animal models will be employed. We will induce dietary copper deficiency during perinatal development in C57BL and C58 mice during several discrete periods from gestation to early adulthood. Repletion studies will be conducted. Copper status during depletion and following repletion will be assessed by determining blood hematocrit and plasma ceruloplasmin activity. Organ (spleen and thymus) and serum levels of total copper and iron will be measured by atomic absorption spectroscopy (AAS). Changes in functional copper pools will be evaluated by assaying the activity of copper-zinc superoxide dismutase or cytochrome c oxidase. Immunological evaluation will include investigating humoral immunity by determining the number of antibody-producing cells in spleen in response to sheep erythrocyte challenge. Cell-mediated immunity will be studied by quantifying mitogen reactivity of cultured thymocytes and splenocytes and by mixed lymphocyte reaction. Lymphocyte subpopulations from spleen will be determined by immuno-fluorescent procedures using monoclonal antibodies to surface antigens. Plant lectins (PNA and SBA) will be used to evaluate maturational development of lymphocytes from thymus, spleen and bone marrow. Serum IgM and IgG will be measured by ELISA techniques. Additional biochemical measurements will be conducted to elucidate the mechanisms of impaired immunity while testing specific hypotheses. Lymphocytes from spleen and thymus will be isolated, and the plasma membrane will be characterized for changes in lipid and protein composition using gas chromatography and gel electrophoresis, respectively. Antioxidant status will be evaluated in lymphoid organs by quantifying specific metabolites. Repletion studies will be conducted comparing cupric or ferrous gluconate and purified mouse ceruloplasmin or transferrin to determine the extent, if any, that altered immunity is iron-dependent. Plasma, brain and spleen levels of norepinephrine will be quantified by HPLC with electrochemical detection. Copper metabolism will be studied in developing lymphoid organs using 67Cu and AAS.