Immunobiological Role of Fertilization Antigen(FA-1)

受精抗原（FA-1）的免疫生物学作用

基本信息

批准号：
7226967
负责人：
RAJESH K NAZ
金额：
$ 24.92万
依托单位：
WEST VIRGINIA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-12-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7226967
关键词：
Acrosome Reaction Active Immunization Adjuvant Affinity Agglutination Amino Acid Sequence Animal Model Animals Antibodies Antibody Formation Antibody Specificity Antigens Bacteriophages Binding Biological Assay Cholera Toxin Class Clinical Clinical Trials Cloning Complementary DNA Contraceptive Agents Contraceptive Vaccines DNA Databases Diagnosis Drug Formulations Enzyme-Linked Immunosorbent Assay Exposure to Family Female Fertility Fertility Rates Fertilization Freund&apos s Adjuvant Funding Gel Glycoproteins Hand Human IgG1 Immobilization Immune response Immunologic Contraception Indirect Fluorescent Antibody Technique Individual Intramuscular Length Libraries Light Lymphocyte Mus Oocytes Pan Genus Peptides Peripheral Phage Display Phosphorylation Polyacrylamide Gel Electrophoresis Polymerase Chain Reaction Procedures Process Proteins Purpose RNA Reaction Recombinants Research Personnel Role Route Seminal Plasma Sepharose Serine Serum Signal Transduction Pathway Site Sodium Dodecyl Sulfate Specificity Sperm Agglutination Sperm Capacitation Spermatogenesis Spermatozoa antibody Staging Structure-Activity Relationship Surface Synthetic Antigens T-Lymphocyte Techniques Technology Testing Testis Therapeutic Threonine Time Tissues Tyrosine Phosphorylation Vaccinated Vaccination Vaccines Western Blotting Woman Zona Pellucida clinical application combinatorial cytotoxicity fertilization antigen human monoclonal antibodies human tissue man men novel peripheral blood programs receptor recombinant peptide sperm cell synthetic peptide vaccine development vaginal fluid vector

项目摘要

DESCRIPTION (provided by applicant): Fertilization antigen (FA-1) is a testis/sperm-specific glycoprotein that is present on sperm of various mammalian species including man and mouse and has a role in fertilization. It is involved in human immunoinfertility and the recent clinical trial indicates that it may have clinical applications in specific diagnosis and treatment of immunoinfertile men. The cDNAs encoding for murine and human FA-1 antigens have been cloned and sequenced. Active immunization of female mice with murine recombinant(r) FA-1 antigen or with another recently identified novel sperm-specific dodecamer synthetic peptide, designated as YLP12, causes an overall 70% reduction, rather than a complete block, of fertility with either vaccine, due to inter-individual variability of the immune response. The present proposal has four specific aims and will focus on obtaining FA-l-reactive human monoclonal antibodies that could be used as a passive immunocontraceptive to avoid variability of the immune response, and on testing the bivalent vaccine having FA-1 antigen and YLP12 together in a single formulation to obtain a complete block in actively immunized female mice. Specific aim 1 is to construct a combinatorial phage display library having y1 heavy (H) and kappa light (L) chains cDNA inserts of Fab antibodies, reverse-transcribed from RNA isolated from peripheral lymphocytes of immunoinfertile men. Specific aim 2 is to select the human rFA-1 antigen-reactive clones from the library using the panning procedure and sequence the Hand L-chain inserts of the selected clones. Specific aim 3 is to investigate the immunobiological effects of the Fab antibodies secreted by the human rFA-1 antigen-reactive clones by using various assays. Specific aim 4 is to investigate the immunocontraceptive effect of the bivalent vaccine, having murine rFA-1 antigen and synthetic YLP12 peptide in a single formulation, in actively immunized female mice. Besides enabling us to study the structure-function relationship, these studies will examine the utility of recombinant FA-1 antigen and synthetic YLP12 peptide, and recombinant human antibodies in immunocontraception and immunoinfertility in humans.

描述（由申请人提供）：受精抗原（FA-1）是一种睾丸/精子特异性糖蛋白，存在于包括人和小鼠在内的各种哺乳动物物种的精子上，并且在受精中起作用。它参与了人类免疫传递性，最近的临床试验表明，它可能在特定的诊断和治疗免疫助力男性中具有临床应用。编码鼠和人FA-1抗原的cDNA已被克隆并测序。用鼠重组（R）FA-1抗原或最近确定的另一种新型的新型精子特异性十二烷合成肽（指定为YLP12）对雌性小鼠进行主动免疫免疫免疫接种，这是由于免疫反应的间个体可变性，这会导致总体上的70％降低70％，而不是完全疫苗的全部疫苗，而不是疫苗。本提案具有四个具体目标，并将集中于获得FA-L反应性人类单克隆抗体，可以用作被动免疫调整，以避免免疫反应的可变性，并测试具有FA-1抗原和YLP12的二价疫苗，以在单个配方中共同获得一个完整的女性群中，以获得单一的配方中，以使其具有活力。具体目的1是构建具有Y1重（H）和Kappa Light（L）链的Fab抗体的组合噬菌体展示库，该插入型Fab抗体的cDNA插入物中，从免疫因素男性的外周淋巴细胞中分离出的RNA反向转录。具体目的2是使用Panning程序从库中选择人RFA-1抗原反应克隆，然后对所选克隆的手L链插入物进行测序。特定目的3是通过使用各种测定法研究由人RFA-1抗原反应克隆分泌的FAB抗体的免疫生物学作用。具体目的4是在主动免疫的雌性小鼠中研究单个配方中的鼠RFA-1抗原和合成YLP12肽的二价疫苗的免疫调节作用。除了使我们能够研究结构功能关系外，这些研究还将研究重组FA-1抗原和合成YLP12肽的实用性，以及在人类免疫传感和免疫接触性中重组人类抗体的实用性。