IMMUNODOMINANT OVARIAN ANTIGENS INVOLVED IN PREMATURE OVARIAN FAILURE

描述(申请人提供)：这项研究计划将表征卵巢早衰(POF)患者血清抗体识别的优势卵巢抗原，目的是更好地了解POF的病理生理机制，并识别用于诊断自身免疫性卵巢疾病的生物标志物。POF患者的血清将被筛选出抗卵巢抗体，然后用于二维凝胶免疫印迹，以识别存在于人、鼠卵巢和卵子提取液中的免疫活性蛋白。将对免疫反应活跃的卵巢和卵子蛋白进行微测序，以确定编码卵巢和卵子抗原的候选基因。通过质谱学获得的多肽微序列将与基因和蛋白质数据库进行比较，以识别自身抗原。将克隆卵巢/卵子自身抗原的全长cDNA，并在细菌和酵母表达系统中表达重组抗原。一个或多个已确定的重组卵靶抗原抗体的发生率将在POF患者和抗卵巢抗体患者中确定。这些研究有望加深对人类卵巢自身免疫的了解，并改进检测抗卵巢抗体的方法，包括检测早期疾病的可能性。对卵巢疾病进行更好的诊断也可能有助于适当的临床随访和选择可能受益于免疫治疗或辅助生殖技术(体外受精/胚胎移植[IVF/ET])以恢复生育能力的患者。

DESCRIPTION (provided by applicant): This research proposal will characterize dominant ovarian antigens recognized by serum antibodies from patients with premature ovarian failure (POF) with the goals of better understanding the pathophysiology of POF and identifying biomarkers for diagnosis of autoimmune ovarian disease. Sera from patients with POF will be screened for antiovarian antibodies and then employed on 2-D gel immunoblots to identify immunoreactive proteins present in human and mouse ovarian and egg extracts. Immunoreactive ovarian and egg proteins will be microsequenced to identify candidate genes encoding ovarian and egg antigens. Peptide microsequences obtained by mass spectrometry will be compared to gene and protein databases to identify autoantigens. Full length cDNAs corresponding to ovarian/egg autoantigens will be cloned and recombinant antigens expressed in bacterial and yeast expression systems. The incidence of antibodies to one or more defined recombinant egg target antigens will then be determined in patients with POF and antiovarian antibodies. The studies are anticipated to lead to a deeper understanding of ovarian autoimmunity in humans and to improved methods of detecting antiovarian antibodies including the possibility of detecting early stage disease. Availability of better diagnostics for ovarian disease may also help in proper clinical follow up and selection of patients who may benefit from immunotherapy or assisted reproductive technologies (in vitro fertilization/embryo transfer [IVF/ET]) to recover their fertility.