FETAL ALCOHOL EFFECTS ON IMMUNE DEVELOPMENT

胎儿酒精综合征（FAS）儿童在宿主防御方面存在缺陷， 危及生命的感染发生率随之增加。 一些患有FAS的儿童在体液和体液免疫方面都有长期的缺陷。 细胞介导的免疫，这表明子宫内暴露于乙醇 可能会干扰免疫系统发育中的关键事件。 该实验室最近的研究表明，在子宫内暴露于 乙醇引起新生小鼠B淋巴细胞发育迟缓。 骨髓和脾脏的表型分析显示， B细胞发育途径中的中间体受到 胎儿酒精暴露 另外，我们注意到一种未知的细胞类型， 在5周龄时，脾B细胞群恢复到 然而，在接近正常水平时，骨髓中的B细胞中间体 在> 5周龄的胎儿乙醇中， 动物 本提案的总体目标是确定 胎儿暴露于乙醇的机制 对B细胞发育中间体的影响。 我们将确定 含乙醇的日粮，（a）其母亲将配对的动物 饲喂对照流质饮食和（B）其母亲将饲喂原种的动物 饮食. 我们将建立定时怀孕，母鼠将被喂养， 适当的液体饮食的各个时期（13 - 19天），这将跨越 准备这些研究中胎儿B细胞的关键期 从妊娠第13 - 19天采集的胎肝和骨髓中 和1日龄小鼠的脾脏，然后每周一次直至6 - 8周龄。 我们将测定不同时期B祖细胞的绝对值 使用流式细胞术和荧光显微镜进行的研究。 B细胞 中间体将通过使用淘选富集在细胞群中， 抗体包被的板和/或通过流式细胞术分选。 The Colony 纯化的祖细胞的形成单位将通过测量 B祖细胞在软琼脂中的克隆性增殖 对选择的生长因子和培养条件的反应。 的 将确定B细胞中间体的发育潜力 通过测量细胞的频率， 分化细胞。 对胎儿乙醇的长期影响将是 通过测试不同年龄的动物的免疫反应来评估， 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彻底的理解 酒精对分化的影响的细胞基础 的B细胞，并提出了潜在的机制，为临床免疫 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.