IMMUNOLOGICAL STUDIES OF THE MALE REPRODUCTIVE SYSTEM
IMMUNOLOGICAL STUDIES OF THE MALE REPRODUCTIVE SYSTEM
批准号:
3323974
负责人:
STEPHEN W BYERS
金额:
$11.96万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1988
资助国家:
美国
项目状态:
已结题
起止时间:
1988-03-01 至 1991-02-28
关键词:
Sertoli cells complementary DNA epididymis epithelium fertility immunology genetic library histochemistry /cytochemistry immunochemistry immunoconjugates immunocytochemistry immunoglobulin A in situ hybridization laboratory mouse laboratory rabbit laboratory rat male reproductive system membrane permeability membrane proteins molecular cloning monoclonal antibody secretory immune system spermatogenesis inhibitor surface antigens tissue /cell culture
中文摘要
睾丸内生殖细胞的发育和成熟精子
附睾会在很大程度上受到全身免疫的保护。
系统。尽管很明显,针对精子表面的抗体
成分可以阻止体外受精,在女性体内,
目前尚不清楚男性体内循环中的抗精子免疫球蛋白
有机会接触到睾丸或附睾的精子。在这
建议我们研究两种不同的方法来解决这个问题
可访问性。我们的第一个具体目标是确认Sertoli和
附睾上皮细胞特异性质膜分子
使用单抗。因为这些细胞紧密联系在一起
参与精子发生和精子成熟
暴露在体循环中,对细胞的识别
它们表面的特定和可获得的决定因素可能会导致,
对我们对这些细胞的了解和
涉及一种选择性损害细胞功能的手段。我们有
最近开发了一种细胞培养系统,在这种系统中我们可以密切地
附睾体内环境的近似性
细胞。使用这些双隔室培养箱,我们可以
现实地评估特定抗体或抗体的效果
细胞功能上的偶联物。随后的细胞特异性抗原
将被提纯并产生针对它们的多克隆抗体。
这些将用于筛选cdna文库和分离抗原。
这些基因随后将被克隆和测序。
提案的最后部分解决了
以另一种方式实现可访问性。众所周知,二聚体IgA,
可以特别地跨多种分泌物跨细胞传播
上皮细胞。尽管这种分子存在于高密度的
精液中的浓度还不知道男性的哪个部位
生殖道参与了它的运输。这个
二聚体IgA可被特异性跨细胞转运的证据
并由生殖道上皮细胞浓缩而成
对于抗精子抗体的传递具有重要意义。
我们建议测量不同疾病患者的分泌型IgA水平
生殖道液,通过定位二聚体IgA受体
免疫细胞化学和原位杂交
通过相关实验研究该分子的细胞转运作用
体外培养的生殖道上皮细胞。
英文摘要
Developing germ cells in the testis and maturing spermatozoa in
the epididymis are largely sheltered from the systemic immune
system. Although it is clear that antibodies against sperm surface
components can prevent fertilization in vitro, and in the female,
it is not clear that circulating anti-sperm IgG in the male will
have access to testicular or epididymal spermatozoa. In this
proposal we investigate two different approaches to this problem
of accessibility. Our first specific aim is to identify Sertoli and
epididymal epithelial cell specific plasma membrane molecules
using monoclonal antibodies. As these cells are intimately
involved in spermatogenesis and sperm maturation and are
exposed to the systemic circulation, the identification of cell
specific and accessible determinants on their surface could lead,
both to significant advantages in our knowledge of these cells and
to a means of selective impairment of cell function. We have
recently developed a cell culture system in which we can closely
approximate the in vivo environment of epididymal or Sertoli
cells. Using these dual compartment culture chambers we can
realistically assess the effects of specific antibodies or antibody
conjugates on cell function. Subsequently cell specific antigens
will be purified and polyclonal antibodies generated against them.
These will be used to screen cDNA libraries and isolate antigen
genes which will then be cloned and sequenced.
The last part of the proposal addresses the problem of
accessibility in another way. It is well known that dimeric IgA,
can be specifically transcytosed across a variety of secretory
epithelia. Although this molecule is present in high
concentrations in semen it is not known which part of the male
reproductive tract are involved in its transport. The
demonstration that dimeric IgA can be specifically transcytosed
and concentrated by reproductive tract epithelial could have
great significance regarding the delivery of anti-sperm antibodies.
We propose to measure levels of secretory IgA in various
reproductive tract fluids, localize the dimeric IgA receptor by
both immunocytochemistry and in situ hybridization and
investigate the transcytosis of this molecule by relevant
reproductive tract epithelia in vitro.
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