注射与浸泡免疫诱导牙鲆免疫应答的比较研究

Injection and immersion are the two routes that commonly taken in vaccination, and the exploration of differences in the immune response of fish induced by these two routes is essential for us to establish a scientific and efficient vaccination strategy. In the present research, based on the previously produced specific antibodies against the T/B lymphocytes, serum and mucosal Ig and polymeric Immunoglobulin Receptor (pIgR), and the previously developed quantitative method for immunogen and the data of the immune response of flounder obtained, The comparative study on the immune response of flounder injected and immersed with different types of antigens was performed concerning the proliferation dynamic of T/B lymphocytes, the response of specific antibodies in serum and mucus, the expression profile of immune-related genes and the antigen uptaking course and Ig transportation by pIgR. Furthermore, the differences in the immune response of flounder induced by injection and immersion were comprehensively analyzed, and the correlation between immune response level and immune effects would be determined. All the research resultant would reveal the differences of immune response characteristics of flounder induced by injection and immersion vaccination at cell, antibody and gene levels, which would provide important data for developing scientific and efficient vaccination strategy.

注射与浸泡是疫苗接种通常采用的两种免疫途径，深入探究两种免疫途径诱导鱼体免疫应答的差异特性，对建立科学、高效的免疫策略具有重要意义。基于前期研制的抗牙鲆T/B淋巴细胞、血清与黏液免疫球蛋白（Ig）及多聚免疫球蛋白受体（pIgR）特异性抗体，以及构建的免疫原摄入定量方法与积累的牙鲆免疫应答资料，比较研究牙鲆在注射与浸泡免疫接种不同类型抗原后T/B淋巴细胞的响应与增殖特性，血清及黏液中特异性抗体应答动态，关键免疫相关基因的差异表达，抗原摄入途径与进程以及pIgR转运动态特征，进而通过综合比较注射与浸泡免疫诱导的牙鲆免疫应答水平差异，分析其与免疫效果之间的关联性；以期从细胞、抗体及基因水平上较为全面而深入地阐明注射与浸泡免疫诱导的牙鲆免疫应答差异特征及其细胞与分子基础，为开展科学高效地免疫接种提供重要理论数据。

注射与浸泡是疫苗接种通常采用的两种免疫途径，深入探究两种免疫途径诱导鱼体免疫应答的差异特性，对建立科学、高效的免疫策略具有重要意义。本项目深入阐明了肌肉注射、腹腔注射与浸泡免疫灭活迟缓爱德华氏菌后牙鲆对菌体抗原的摄入动态过程，并掌握各免疫组织中抗原递呈相关基因的应答表达差异特征，发现菌苗剂量、免疫途径和浸泡时间对各组织的抗原摄入量以及摄入动态具有显著影响；利用研制的我国海水养殖鱼类重要病毒各类型候选疫苗免疫牙鲆，阐明了注射免疫、浸泡免疫以及口服免疫疫苗后牙鲆T/B淋巴细胞亚群响应动态、特异性抗体分泌特点以及免疫相关基因的表达特征，发现注射免疫相比浸泡和口服可引起更高的免疫应答水平和免疫保护效果，而浸泡和口服免疫则会引起皮肤、鳃和肠道等黏膜免疫相关组织更迅速免疫响应。明晰了包括IL-2、IL-6和G-CSF在内的多种细胞因子的佐剂效应特征，发现它们与亚单位疫苗共免疫后可显著增强T/B淋巴细胞的增殖、抗体分泌及免疫相关基因表达，提高免疫应答水平，最终提升疫苗保护效果。检测了鲆鲽鱼类IgT与pIgR/pIgRL在浸泡和注射免疫后响应特征，发现它们高表达与黏膜相关组织，在浸泡免疫后可显著诱导牙鲆IgT和pIgR/pIgRL应答表达。研究结果从细胞、蛋白和分子水平揭示了浸泡免疫和注射免疫在免疫应答特性以及免疫应答效果间的差异机制，为海水养殖鱼类高效免疫策略提供了重要的理论基础。