FETAL ALCOHOL EFFECTS ON IMMUNE DEVELOPMENT

胎儿酒精综合征(FAS)儿童存在宿主防御缺陷和 随之而来的是威胁生命的感染发生率的增加。 一些患有自闭症的儿童在体液和体液两方面长期存在缺陷 细胞免疫，表明宫内接触乙醇 可能会干扰免疫系统发育的关键事件。 该实验室最近的研究表明，在子宫内暴露于 乙醇可抑制新生小鼠B淋巴细胞的发育。 骨髓和脾的表型分析显示，几个 B细胞发育途径中的中间产物受 胎儿接触酒精。此外，我们注意到一种未知的细胞类型， 在五周龄时，脾B细胞数量恢复到 然而，在接近正常水平的情况下，B细胞在骨髓中居中 胎儿酒精在5周龄时仍低于正常值 动物。这项提案的总体目标是确定 胎儿酒精暴露的机制子宫乙醇暴露 B细胞发育中间体。我们将确定 在含有乙醇的传统中，(A)母亲将配对的动物 饲喂对照流质饲料和(B)其母亲将饲喂家畜的动物 节食。我们将安排定时怀孕，大坝将被喂食 适合不同时期(13-19天)的流质饮食 将在这些研究中准备胎儿B细胞的临界期 取自妊娠13-19天的胎儿肝脏和骨髓 1天龄小鼠的脾和6-8周龄的小鼠的脾。 我们将在不同的阶段测定B细胞祖细胞的绝对数量 用流式细胞仪和荧光显微镜观察细胞发育情况。B细胞 中间体将通过使用淘洗来丰富细胞群 抗体包被平板和或用流式细胞术分选。殖民地 纯化的祖细胞的形成单位将通过测量来确定 B细胞祖细胞在软琼脂中的克隆性增殖 对选定的生长因素和培养条件作出反应。这个 B细胞中间体的发展潜力将被确定 通过测量细胞的频率可以产生更多的 分化的细胞。对胎儿酒精的长期影响将是 通过测试不同年龄动物对 1型和2型T非依赖性抗原和T依赖性抗原。 完成这些实验应该提供一个透彻的理解 酒精对分化的影响的细胞学基础 并提示临床免疫的潜在机制 与FAE相关的抑制。

Fetal alcohol syndrome (FAS) children have defects in host defense and concomitant increase in the incidence of life threatening infections. Some children with FAS have long lasting deficiencies in both humoral and cell mediated immunity, suggesting that intrauterine exposure to ethanol may interfere with critical events in development of the immune system. Recent studies from this laboratory have shown that in utero exposure to ethanol caused a retarded development of B lymphocytes in neonatal mice. Phenotypic analysis of bone marrow and spleen showed that several intermediates in the B cell development pathway were affected by the fetal ethanol exposure. It addition, we noted that an unknown cell type, with an five weeks of age the splenic B cell population had returned to near normal levels, however, the B cell intermediates in the bone marrow remained below normal values at >5 weeks of age in the fetal ethanol animals. The overall goal of this proposal is to determine the mechanisms by which fetal ethanol exposure uterine exposure to ethanol on B cell developmental intermediates. We will determine the effects of intradiet containing ethanol, to (a) animals whose mothers will be pair fed a control liquid diet and (b) animals whose mothers will fed a stock diet. We will set up timed pregnancies and the dams will be fed the appropriate liquid diet for various periods (13-19 days), which will span the critical period of fetal B cell in these studies will be prepared from fetal liver taken at days 13-19 of gestation, and from bone marrow and spleens of mice 1 day old and then weekly through 6-8 weeks of age. We will determine the absolute of B cell progenitors at different stages of development using flow cytometry and fluorescence microscopy. B cell intermediates will be enriched in cell population by using panning on antibody coated plates and.or sorting by flow cytometry. The colony forming units of the purified progenitors will be determine by measuring the clonal proliferation of the B cell progenitors in soft agar in response to selected growth factors and culture conditions. The developmental potential of the B cell intermediates will be determined by measuring the frequency of cells that can give rise to a more differentiated cell. The long term effects on fetal ethanol will be assessed by testing the immune response of animals of different ages to Type-1 and -2 T independent antigens and to T dependent antigens. Completion of these experiments should provide a thorough understanding of the cellular basis for the effects of alcohol on the differentiation of B cells and suggest potential mechanisms for the clinical immune suppression associated with FAE.