巨型艾美耳球虫激发CD8+T细胞应答的免疫优势抗原分析

Our previous studies showed that dominant CD8+ T cells existed in Eimeria maxima immunized chickens, which supports our current hypothesis that immunodominant antigens of CD8+ T cells exist in E. maxima. In this study, E. maxima cDNA expression library is used to transfect transformed chicken dendritic cells, which in turn induce the release of IFN-γ from specific CD8+ T cells derived the same bird immunized and infected with E. maxima. The immunodominant antigens will be explored from the cDNA library pools which inducted the highest level of IFN-γ. After several rounds of repeat tests, we will identify immunodominant antigens for E. maxima. We will also be able to varify the antigens characteristics as well as the potential of them as vaccine candidates. The single-bird based methodology is novel to provide a non-inbred chicken model for the current research work, and to an extension, for the research of avian immunology. After finishing this project, two to four graduate students will be trained, at least 3 international articles will be publlished and 1 patent will be applied.

我们在前期研究中已证实巨型艾美耳球虫感染的鸡CD8+T细胞中出现优势性群体，为我们解析巨型艾美耳球虫CD8+T细胞免疫优势抗原（immunodominant antigens）提供了有力证据。基于此，本研究将利用巨型艾美耳球虫cDNA表达文库转染的树突状细胞系刺激同一实验鸡（经巨型艾美耳球虫免疫并攻毒感染）来源的外周血CD8+T细胞，通过IFN-γ释放实验测定IFN-γ，对IFN-γ释放量最高者对应的文库部分继续进行第二轮转染树突状细胞，通过3-5轮筛选而获得巨型艾美耳球虫激发CD8+T细胞免疫优势抗原。本研究还将对其特征及其作为疫苗候选进行研究。利用同一实验鸡进行巨型艾美耳球虫免疫优势抗原的研究这一技术上的创新，将为禽免疫学研究提供非近交系动物模型。本研究将培养研究生2-4名，发表SCI文章3篇以上，申请专利1项。

巨型艾美耳球虫是严重危害养鸡业的7 种艾美耳球虫中免疫原性最强的一种。巨型艾美耳球虫本身是否存在具有保护性的免疫优势抗原及其在疫苗开发中的应用是本研究的核心内容。我们通过构建巨型艾美耳球虫特异性的cDNA均一化表达文库，制备针对巨型艾美耳球虫AMA1抗原的单抗，验证巨型艾美耳球虫的潜在优势抗原AMA1和IMP1异源表达后对宿主提供的针对巨型艾美耳球虫的免疫保护，测定特异性CD8+T细胞的CDR3序列并进行分析，初步阐述了巨型艾美耳球虫免疫原性强的免疫学机制，为进一步的深入研究打下了良好基础。发表研究相关的SCI论文6篇（其中第一标注3篇，通讯作者4篇）；国际会议摘要2篇，国内会议论文摘要3篇，培养毕业博士研究生1名，硕士研究生3名，申请并获国家专利授权1项。