IN VITRO MODEL OF POLARIZED TROPHOBLAST

极化滋养细胞的体外模型

基本信息

批准号：
2067164
负责人：
JOSEPH DANCIS
金额：
$ 24.28万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-09-30 至 1994-12-31
项目状态：
已结题

项目摘要

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.

将开发一种滋养细胞的体外模型，这将允许研究胎盘功能的许多方面孕产妇艾滋病毒感染。滋养细胞将保持在组织中在双色室中的文化，可独立进入顶端滋养细胞的（母体）和基底（胎儿）表面。标准为了成功准备，将包括屏障功能的测试（电阻，肌蛋白和其他的限制转移分子，连续单层的微观证据）和测试极性（分化为EM上的顶端和基底外侧结构域，定向挤出病毒体并优先吸收转铁蛋白顶表）。该模型将使用三种单元格类型：从学期和从一个孕期胎盘。这些模型将经过测试旨在研究其两个主要角色：养分的转移胎儿以及激素的合成和分泌维持怀孕。抗HIV药物的潜在毒性将通过将滋养细胞暴露于药物中，然后重新测试其功能。艾滋病毒药物的转移和代谢也将是研究利用模型的极性。同时模型在研究免疫学问题的有用性将探讨有关艾滋病和其他产前感染的信息。这 FC受体的表达和IgG的旋转转移将会可以测量。通过将FC受体引入滋养细胞如果重组技术将进行扩增，以扩大此功能必要的。该模型将应用于与之相关的问题胎儿中疾病的表现，例如特定的转移抗体，病毒抗原和感染剂以及复合物的抗体抗原和抗体。