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
睾丸中生殖细胞的发育和精子的成熟
附睾在很大程度上不受全身免疫系统的影响,
系统 虽然很明显,精子表面的抗体
成分可以防止体外受精,在女性中,
目前尚不清楚男性体内循环的抗精子IgG是否会
能够接触到睾丸或附睾精子 在这
建议我们调查两种不同的方法来解决这个问题
无障碍环境。 我们的第一个具体目标是确定塞尔托利和
附睾上皮细胞特异性质膜分子
使用单克隆抗体。 由于这些细胞与
参与精子发生和精子成熟,
暴露于体循环,细胞的鉴定
其表面上特定和可获得的决定因素可能导致,
我们对这些细胞的了解都有很大的优势,
涉及一种选择性损害细胞功能的方法。 我们有
最近开发了一种细胞培养系统,
接近附睾或Sertoli的体内环境
细胞 使用这些双室培养室,我们可以
真实地评估特定抗体或抗体的影响
对细胞功能的影响。 随后细胞特异性抗原
将被纯化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
这些基因将被克隆和测序。
建议的最后一部分涉及以下问题:
另一种方式的可访问性。 众所周知,二聚伊加,
可以特异性地通过多种分泌型
上皮细胞 虽然这种分子存在于高
精液中的浓度,目前还不知道男性的哪一部分,
生殖道参与其运输。 的
证明二聚体伊加可以特异性转胞吞,
并被生殖道上皮细胞聚集
对于抗精子抗体的递送具有重要意义。
我们建议测量各种组织中分泌伊加的水平,
生殖道液体,本地化二聚体伊加受体,
免疫细胞化学和原位杂交,
研究该分子的转胞吞作用,
体外生殖道上皮细胞。
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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