IN VITRO MODEL OF POLARIZED TROPHOBLAST

将开发一种滋养层细胞的体外模型，这将使 胎盘功能的多方面研究 母体感染艾滋病毒。滋养层细胞将在组织中维持 在两室室中培养，提供独立的根尖通路 滋养细胞的(母体)和基底部(胎儿)表面。标准 成功的准备将包括屏障功能的测试 (电阻、菊粉限制转移及其他 分子，连续单层的微观证据)和测试 极性(在EM上分化为顶区和基底侧区， 病毒粒子的定向排泄和转铁蛋白的优先摄取 顶面)。模型将使用三种单元格类型： 足月分离转化滋养层细胞和原代滋养层细胞 取自妊娠早期的胎盘。这些模型将接受测试 旨在研究它的两个主要作用：营养物质的转移 对胎儿和激素的合成和分泌的影响 维持妊娠。抗艾滋病毒药物的潜在毒性将 通过将滋养层细胞暴露在药物中，然后重新测试其 功能。艾滋病毒药物的转移和代谢也将是 利用模型的极性进行了研究。同时， 该模型在免疫学问题研究中的应用 关于艾滋病和其他产期感染，将进行探讨。这个 Fc受体的表达与免疫球蛋白的跨上皮转移 被测量。Fc受体导入滋养层细胞的研究 如果出现以下情况，将采用重组技术来放大这一功能 这是必要的。该模型将应用于与 表现为疾病在胎儿体内的转移等特异性 病毒抗原和感染性病原体的抗体以及病毒的复合体 抗原和抗体。

An in vitro model of trophoblast will be developed which will permit research into many aspects of placental function as it relates to maternal HIV infection. The trophoblast will be maintained in tissue culture in a bicameral chamber providing independent access to the apical (maternal) and basal (fetal) surfaces of the trophoblast. The criteria for a successful preparation will include tests of barrier function (electrical resistance, restricted transfer of inulin and other molecules, microscopic evidence of a continuous monolayer) and tests of polarity (differentiation into apical and basolateral domains on EM, directed extrusion of virions and preferential uptake of transferrin from the apical surface). Three cell types will be used for the model: transformed trophoblast and primary trophoblast isolated from term and from first trimester placenta. The models will be subjected to tests designed to investigate two of its major roles: the transfer of nutrients to the fetus and the synthesis and secretion of hormones for the maintenance of pregnancy. The potential toxicity of anti-HIV drugs will be explored by exposing trophoblast to the drug and then re-testing its function. The transfer and metabolism of the HIV drugs will also be studied taking advantage of the polarity of the model. Concurrently the usefulness of the model for the study of immunological questions concerning AIDS and other intrapartum infections will be explored. The expression of Fc receptors and the transepithelial transfer of IgG will be measured. Introduction of Fc receptors into trophoblast by recombinant techniques will be undertaken to amplify this function if necessary. The model will be applied to questions pertinent to the manifestation of disease in the fetus such as the transfer of specific antibodies, of viral antigens and infectious agents and of complexes of antigens and antibodies.