ZONA PELLUCIDA: IMMUNOPATHOLOGIC STUDY

透明带：免疫病理学研究

• 批准号: 2200467
• 负责人: KENNETH S.K. TUNG
• 金额: $ 9.78万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2200467
• 关键词: B lymphocyte; T lymphocyte; autoimmune disorder; clone cells; contraceptives; cytokine; disease /disorder model; egg /ovum; enzyme linked immunosorbent assay; epitope mapping; female; fertility immunology; genetic library; immunization; immunopathology; laboratory mouse; molecular cloning; monoclonal antibody; ovary disorder; western blottings; B lymphocyte; T lymphocyte; autoimmune disorder; clone cells; contraceptives; cytokine; disease /disorder model; egg /ovum; enzyme linked immunosorbent assay; epitope mapping; female; fertility immunology; genetic library; immunization; immunopathology; laboratory mouse; molecular cloning; monoclonal antibody; ovary disorder; western blottings

In the proposal, we will elucidate the mechanisms, and subsequently the therapeutic approach to human ovarian autoimmune disease, a known cause of premature ovarian failure. To do this, we will use a new model of murine autoimmune oophoritis which we have developed, by immunizing mice with a synthetic 13-me peptide. ZP3330-342 from ZP3 of the murine zona pellucida. With T cell clones that can adoptively transfer autoimmune oophoritis, we will map the pathogenic epitopes in the ZP3 peptide and determine the critical cytokines responsible for the ovarian disease. The model also will be exploited to determine the feasibility of a contraceptive vaccine based on ZP3. The unique ZP3330-342 peptide contains both B cell (ZP3336-342) and T cell epitopes that readily induce antibody and T cell responses to ZP3. Antibody to the ZP3. Antibody to the ZP3 peptide results in reversible infertility, whereas the ZP3- specific T cells cause autoimmune oophoritis and ovarian destruction. Although an immunogen that induces an antibody response without eliciting a concomitant T cell response to ZP3 will likely be a safe ZP3 vaccine, we need first to show that such an antibody per se does not cause autoimmune oophoritis. Our strategy in investigating the critical role of antibody in autoimmune oophoritis will be to synthesize immunogens that contain the B cell epitope of ZP3, coupled to either a foreign antigen or a peptide capable of stimulating a strong T cell response. When the immunogen has been shown not to stimulate a T cell response to ZP3, we will study it for induction of: 10 infertility and reversal, 2) abnormal ovarian response to gonadotrophin and 3) oophoritis in female (C57BL/6xA/J)F1(B6AF1)mice. Our second approach will involve the transfer of saturating quantities of antibodies to ZP3 into normal female B6AF1 mice and study their fertility, ovarian function and ovarian pathology. We will investigate two kinds of antibodies that react with the native determinants of ZP3: 1) monoclonal antibodies to the B cell epitope(s) of ZP3330-342, 2) monoclonal antibodies to determinants of ZP3 outside the ZP3 outside the ZP3 330-342 peptide. The second group of antibodies will be produced by normal female B6AF1 mice immunized with the ZP3 330-339 peptide which lacks an intact B cell epitope. Although the ZP3 330-339 peptide does not elicit an antibody response to ZP3330- 342, it induces a T cell response to ZP3330-339 and autoimmune oophoritis. Remarkable, we have accrued evidence that the endogenous ovarian antigens from the injured ovaries in these mice stimulate an antibody response to the ZP3 determinants outside the peptide ZP3 330- 342, and such antibodies react with the zone pellucida in vivo. Because of the important implications that the new phenomenon of "antibody amplification' has on autoimmune oophoritis and autoimmunity in general, we will investigate the structure of the ZP3 determinants recognized by the amplified antibodies, and the biological significance of the "antibody amplification" phenomenon.

在提案中，我们将阐明机制，然后 人卵巢自身免疫性疾病的治疗方法，这是已知原因 早产卵巢衰竭。 为此，我们将使用一个新的模型 通过免疫小鼠，我们已经开发出的鼠自身免疫性卵泡 与合成13-ME肽。 ZP3的ZP3330-342 Pellucida。 用T细胞克隆可以采用自身免疫性 卵形炎，我们将绘制ZP3肽中的病原体表位和 确定导致卵巢疾病的关键细胞因子。 该模型还将被利用以确定 基于ZP3的避孕疫苗。 独特的ZP3330-342肽 同时包含B细胞（ZP3336-342）和T细胞表位，很容易诱导 抗体和T细胞对ZP3的反应。 ZP3的抗体。 抗体 ZP3肽会导致可逆的不育，而ZP3- 特定的T细胞会引起自身免疫性卵巢炎和卵巢破坏。 虽然一种免疫原质，可引起抗体反应而不引起抗体反应 伴随T细胞对ZP3的响应可能是一种安全的ZP3疫苗， 我们首先需要证明这种抗体本身不会引起 自身免疫性卵巢炎。 我们调查关键作用的策略 自身免疫性卵巢炎的抗体将是合成免疫原的 包含ZP3的B细胞表位，耦合到任何一个外国 抗原或能够刺激强T细胞反应的肽。 当显示免疫原不刺激T细胞反应时 ZP3，我们将研究它以诱导：10种不育症和逆转，2） 卵巢异常对促性腺营养蛋白的反应和3）雌性卵形炎 （C57BL/6XA/J）F1（B6AF1）小鼠。 我们的第二种方法将涉及 将饱和量的抗体转移到ZP3中 B6AF1小鼠并研究其生育能力，卵巢功能和卵巢 病理。 我们将研究两种与 ZP3的天然决定因素：1）对B细胞的单克隆抗体 ZP3330-342的表位，2）ZP3决定因素的单克隆抗体 在ZP3外面的ZP3 330-342肽之外。 第二组 抗体将由普通雌性B6AF1小鼠产生 ZP3 330-339肽缺乏完整的B细胞表位。 虽然 ZP3 330-339肽不会引起对ZP3330-的抗体反应 342，它诱导了T细胞对ZP3330-339和自动免疫的响应 卵形炎。 引人注目的是，我们有证据表明内源性 这些小鼠的受伤卵巢的卵巢抗原刺激了 对肽ZP3 330-之外ZP3决定因素的抗体反应 342，此类抗体与体内的质体区反应。 因为 “抗体的新现象”的重要含义 放大对自身免疫性卵形炎和自身免疫的具有 我们将研究由ZP3决定因素的结构 放大的抗体及其生物学意义 “抗体扩增”现象。