Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, China.
Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China.
Fish Shellfish Immunol. 2019 Jun;89:393-402. doi: 10.1016/j.fsi.2019.04.022. Epub 2019 Apr 11.
In order to investigate the dynamic distribution of antigen in different tissues post vaccination, an absolute real-time quantitative PCR was employed to detect the amount of antigen in flounder (Paralichthys olivaceus) post intraperitoneal (i.p.) injection with three concentrations (10, 10, 10 CFU ml) of formalin-inactivated Edwardsiella tarda bacterin. The results showed that the amount of uptaken antigen quickly increased and then decreased in different tissues. The peak occurred first in the spleen and head kidney at 6-9 h after injection, and in the liver and blood at 9-15 h, then in the gill, intestine and skin at 15-24 h, finally in the muscle at 24-36 h. The amount of antigen was highest in the spleen and head kidney, followed by the blood, liver and gill, and lowest in the intestine, skin and muscle. Among the three concentration groups, the amount of antigen increased with the increasing concentration of the vaccine in the blood, liver, gill, intestine, skin and muscle, except for the spleen and head kidney, in which more antigens were found in the 10 CFU ml group than that in 10 CFU ml group. Moreover, IIFA and western blotting was performed to examine the tissue distribution of antigen at 9 h after vaccination with 10 CFU ml formalin-inactivated E. tarda. The bacteria were mainly observed in the spleen and head kidney, then the liver, gill and blood, and least in the intestine, skin and muscle, which was roughly in accordance with the results of absolute qPCR. Furthermore, the expressions of CD4-1, MHC IIα, CD8α and MHC Iα in different tissues were detected by RT-qPCR, and the expression levels of these genes were highest in the spleen and head kidney, then in the blood, gill, liver, and lowest in the intestine, skin and muscle. All these results provided useful information for dynamic transportation of antigen uptake post vaccination, and also deepened the understanding of immune response to the injection vaccination.
为了研究疫苗接种后抗原在不同组织中的动态分布，本研究采用绝对实时定量 PCR 检测了福尔马林灭活迟钝爱德华氏菌菌苗经腹腔注射（i.p.）后牙鲆（Paralichthys olivaceus）不同组织中抗原的含量。结果表明，不同组织中摄取的抗原量迅速增加，然后减少。在注射后 6-9 小时，抗原量首先在脾脏和头肾中达到峰值，然后在肝脏和血液中在 9-15 小时达到峰值，然后在鳃、肠和皮肤中在 15-24 小时达到峰值，最后在肌肉中在 24-36 小时达到峰值。在脾脏和头肾中，抗原量最高，其次是血液、肝脏和鳃，在肠、皮肤和肌肉中最低。在三个浓度组中，除了脾脏和头肾，随着疫苗浓度的增加，血液、肝脏、鳃、肠、皮肤和肌肉中抗原的含量均增加，而在 10 个 CFU/ml 组中发现更多的抗原。此外，在接种 10 个 CFU/ml 福尔马林灭活 E. tarda 后 9 小时，采用 IIFA 和 Western blot 检测了抗原在组织中的分布。细菌主要在脾脏和头肾中观察到，然后在肝脏、鳃和血液中观察到，在肠、皮肤和肌肉中观察到的细菌最少，这与绝对 qPCR 的结果大致相符。此外，通过 RT-qPCR 检测了不同组织中 CD4-1、MHC IIα、CD8α 和 MHC Iα 的表达，这些基因的表达水平在脾脏和头肾中最高，其次是血液、鳃、肝脏，在肠、皮肤和肌肉中最低。这些结果为疫苗接种后抗原摄取的动态运输提供了有用的信息，也加深了对注射疫苗免疫反应的理解。
