Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, China.
Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266235, China.
Fish Shellfish Immunol. 2018 Jun;77:374-384. doi: 10.1016/j.fsi.2018.04.009. Epub 2018 Apr 5.
Cytokines play vital roles in mounting immune responses and activating host defense network. In this study, the expression plasmid pcDNA3.1 (pcN3) encoding four flounder (Paralichthys olivaceus) cytokines including IL-1β, TNF-α, IL-8 or G-CSF (pcIL-1β, pcTNF-α, pcIL-8 and pcG-CSF) were successfully constructed, and their adjuvant potential on an Edwardsiella tarda (E. tarda) subunit vaccine OmpV (rOmpV) were comparatively analyzed in vaccinated flounder model. Results revealed that flounder vaccinated with rOmpV plus pcIL-1β, pcIL-8 or pcG-CSF produced the relative percent survivals (RPS) of 71%, 65% and 49% respectively, which were higher than that in flounder vaccinated with rOmpV plus pcTNF-α (39%) or pcN3 (36%, the control group). Immunological analysis showed that: (1) except pcTNF-α, higher levels of anti-E. tarda serum antibodies and sIg + lymphocytes in spleen, head kidney and peripheral blood were significantly enhanced by pcIL-1β, pcIL-8 or pcG-CSF, however, pcIL-8 and pcIL-1β enhanced higher levels of sIg + lymphocytes and anti-E. tarda antibodies than pcG-CSF; (2) pcTNF-α could promote the up-regulation of genes participated in cellular immunity (MHCIα, IFN-γ, CD8α and CD8β), pcIL-1β could enhance the expression of genes related to humoral immunity (CD4-1, CD4-2, MHCIIα and IgM), and all the detected genes were augmented by pcIL-8 and pcG-CSF; Among the four cytokines, pcIL-8 and pcIL-1β could strengthen the highest levels of genes participated in cellular immunity and humoral immunity, respectively. These results demonstrated that pcIL-8 and pcIL-1β could enhance stronger cellular and/or humoral immunity induced by rOmpV than pcG-CSF and pcTNF-α, and evoked higher RPS against E. tarda challenge in flounder, which indicated that pcIL-8 and pcIL-1β are promising adjuvants of vaccines in controlling E. tarda infection.
细胞因子在启动免疫反应和激活宿主防御网络方面发挥着重要作用。在这项研究中，成功构建了编码四种牙鲆（Paralichthys olivaceus）细胞因子（IL-1β、TNF-α、IL-8 或 G-CSF）的表达质粒 pcDNA3.1（pcN3）（pcIL-1β、pcTNF-α、pcIL-8 和 pcG-CSF），并在牙鲆接种模型中比较分析了它们在爱德华氏菌（Edwardsiella tarda）亚单位疫苗 OmpV（rOmpV）中的佐剂潜力。结果表明，与接种 rOmpV 加 pcTNF-α（39%）或 pcN3（对照组，36%）的牙鲆相比，接种 rOmpV 加 pcIL-1β、pcIL-8 或 pcG-CSF 的牙鲆的相对存活率（RPS）分别为 71%、65%和 49%。免疫分析表明：（1）除 pcTNF-α 外，pcIL-1β、pcIL-8 或 pcG-CSF 可显著增强抗爱德华氏菌血清抗体和脾脏、头肾和外周血中 sIg+淋巴细胞的水平，但 pcIL-8 和 pcIL-1β 增强的 sIg+淋巴细胞和抗爱德华氏菌抗体水平高于 pcG-CSF；（2）pcTNF-α 可促进参与细胞免疫的基因（MHCIα、IFN-γ、CD8α 和 CD8β）的上调，pcIL-1β 可增强与体液免疫相关的基因（CD4-1、CD4-2、MHCIIα 和 IgM）的表达，所有检测到的基因均由 pcIL-8 和 pcG-CSF 增强；在这四种细胞因子中，pcIL-8 和 pcIL-1β 分别能增强细胞免疫和体液免疫相关基因的表达水平。这些结果表明，pcIL-8 和 pcIL-1β 可增强 rOmpV 诱导的细胞和/或体液免疫，比 pcG-CSF 和 pcTNF-α 更强，并在牙鲆中引发更高的针对爱德华氏菌挑战的 RPS，表明 pcIL-8 和 pcIL-1β 是控制爱德华氏菌感染的疫苗有前途的佐剂。
