IMMUNOBIOLOGICAL ROLE OF FERTILIZATION ANTIGEN FA-1

受精抗原 FA-1 的免疫生物学作用

• 批准号: 2888966
• 负责人: RAJESH K NAZ
• 金额: $ 26.45万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2888966
• 关键词: SDS polyacrylamide gel electrophoresis; active immunization; chromosomes; enzyme linked immunosorbent assay; fertility immunology; gene expression; glutathione transferase; human genetic material tag; human tissue; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rabbit; molecular cloning; monoclonal antibody; northern blottings; nucleic acid sequence; polyadenylate; recombinant proteins; restriction mapping; sperm; sperm capacitation; surface antigens; vaccine development; zona pellucida glycoproteins

DESCRIPTION: (adapted from the applicants abstract) Fertilization antigen (FA-1) is a tissue-specific glycoprotein that is present on sperm of various mammalian species including mouse and man. It has a function in sperm capacitation/acrosome reaction, has a receptor/ligand activity for ZP3 of oocyte zona pellucida (ZP), and has a definite role in immunoinfertility in humans. Human cognate FA-1 antigen undergoes tyrosine phosphorylation and autophosphorylation during capacitation/acrosome reaction and ZP binding. cDNA encoding for murine FA-1 antigen has been cloned and sequenced, and was found to be a novel gene having testis-specific expression, and antibodies to the recombinant murine antigen completely blocked sperm-ZP binding in vitro (Proc Natl Acad Sci USA, 94:4704-4709, 1997). The present proposal has five specific aims. Specific Aim #1 is to extend the in vitro studies to examine the immunocontraceptive effects of the murine recombinant FA-1 antigen in actively-immunized female mice. Specific aim 2 is to clone and sequence the human homologue of the murine FA-1 gene. Antibodies (monospecific polyclonal/monoclonal) against the human cognate FA-1 antigen and the murine FA-1 cDNA will be used as probes to screen the human testes cDNA-lgt11 lZAPiilgt 10 libraries. The specific clones will be subjected to restriction mapping and investigated for homology with any known sequence. Specific aim 3 is to investigate the tissue-specific expression of the human FA-1 gene using Northern blot and RT-PCR procedures. Specific aim 4 is to study the chromosomal localization of the human FA-1 gene. Specific aim 5 is to investigate the immunobiological effects of the human recombinant FA-1 antigen. The human recombinant antigen will be obtained by using the glutathione S-transferase fusion system and examined for its interaction with the human ZP3 and for its involvement in immunoinfertility, and its antibodies will be investigated for their effects on sperm function. Besides enabling the PI to study the structure-function relationship, these studies will help them investigate the potential utility of the human recombinant FA-1 antigen is immunocontraception and immunoinfertility in humans.

描述：（改编自申请人摘要）施肥抗原 （FA-1）是一种组织特异性糖蛋白，存在于各种精子上 哺乳动物物种，包括小鼠和人。 它在精子中具有功能 电容/ACROSOM体反应，具有ZP3的受体/配体活性 卵母细胞Zona pellucida（ZP），在免疫诱导中具有明确的作用 人类。 人类同源FA-1抗原经历酪氨酸磷酸化和 在电容/ACROSOM反应和ZP结合期间的自磷酸化。 鼠FA-1抗原编码的cDNA已被克隆和测序，并且是 发现是具有睾丸特异性表达和抗体的新基因 重组鼠抗原完全阻止了精子-ZP结合 体外（Proc Natl Acad Sci USA，94：4704-4709，1997）。 目前的提议 具有五个具体目标。 特定目的＃1是扩展体外研究 检查鼠重组FA-1的免疫调节作用 主动免疫的雌性小鼠中的抗原。 具体目标2是克隆和 序列的鼠FA-1基因的人类同源物。 抗体 （单特异性多克隆/单克隆）针对人类同源FA-1抗原 鼠FA-1 cDNA将用作筛选人睾丸的探针 cDNA-LGT11 lzapiilgt 10库。 特定克隆将受到 限制映射并使用任何已知序列进行了研究。 特定目的3是研究人类的组织特异性表达 FA-1基因使用北印迹和RT-PCR程序。 具体目标4是 研究人FA-1基因的染色体定位。 具体目标5 是研究人类重组FA-1的免疫生物学作用 抗原。 人类重组抗原将通过使用 谷胱甘肽S-转移酶融合系统并检查其相互作用 与人类的ZP3及其参与免疫促性及其有关 将研究其对精子功能的影响。 除了使PI能够研究结构功能关系外， 研究将帮助他们研究人类的潜在效用 重组FA-1抗原是免疫感应和免疫接触性的 人类。