Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, PR China.
Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, No. 1 Wenhai Road, Aoshanwei Town, Qingdao, PR China.
Vaccine. 2018 Feb 28;36(10):1316-1322. doi: 10.1016/j.vaccine.2017.11.074. Epub 2018 Feb 1.
In our previous study, ten candidate proteins have been identified with immunogenicity and protection against Vibrio anguillarum in flounder (Paralichthys olivaceus). Among them, FlaC is the important outer protein in the flagellum with immunogenicity; VAA, OmpK and OmpR are protection proteins against V. anguillarum. In this paper, FlaC supplemented with VAA, OmpK or OmpR as bivalent subunit vaccine candidates, and their immune response of flounder and protective effects were evaluated, respectively. Recombinant(r) proteins of FlaC were mixed with rVAA, rOmpK and rOmpR, respectively, rVAA + rFlaC (AF), rOmpK + rFlaC (KF) and rOmpR + rFlaC (RF); formalin-killed cells (FKC) or PBS were injected to flounder, respectively. After immunization, the percentages of CD3 T lymphocytes and surface membrane immunoglobulin-positive (sIg) B lymphocytes in peripheral blood lymphocytes (PBLs), total antibodies (TA), specific antibodies against V. anguillarum (VA), specific antibodies against bivalent recombinant proteins (PA), the expression of immune-related genes and relative percent survivals (RPS) were measured, respectively. The results showed that three bivalent vaccines candidates and FKC could induce the proliferation of sIg B lymphocytes and CD3 T lymphocytes in PBLs. The TA, VA and PA induced in bivalent vaccines candidates and FKC groups were significantly higher than that of the control group. CD3, IgM, CD4-1, CD4-2, CD8α and CD8β genes were up-regulated. After challenge with V. anguillarum, RPS in AF, KF, RF and FKC groups exhibited 62.6 ± 2.33%, 78.95 ± 3.01%, 75.45 ± 0.97%, and 56.71 ± 2.15% respectively. The results revealed that three bivalent vaccines candidates and FKC could induce the immune response in flounder, and have good protection against V. anguillarum, and KF can be an efficient bivalent subunit vaccine candidate.
在我们之前的研究中，已经鉴定出十种对鳗弧菌具有免疫原性和保护作用的候选蛋白。其中，FlaC 是鞭毛中的重要外膜蛋白，具有免疫原性；VAA、OmpK 和 OmpR 是抗鳗弧菌的保护蛋白。本文以 FlaC 为载体，分别与 VAA、OmpK 或 OmpR 组成二价亚单位疫苗候选物，评价其对牙鲆的免疫应答和保护效果。将 FlaC 的重组蛋白分别与 rVAA、rOmpK 和 rOmpR 混合，制成 rVAA+rFlaC（AF）、rOmpK+rFlaC（KF）和 rOmpR+rFlaC（RF）；分别将福尔马林灭活细胞（FKC）或 PBS 注射到牙鲆体内。免疫后，检测外周血淋巴细胞（PBL）中 CD3 T 淋巴细胞和表面膜免疫球蛋白阳性（sIg）B 淋巴细胞的百分比、总抗体（TA）、抗鳗弧菌的特异性抗体（VA）、抗二价重组蛋白的特异性抗体（PA）、免疫相关基因的表达和相对存活率（RPS）。结果表明，三种二价疫苗候选物和 FKC 均可诱导 PBL 中 sIg B 淋巴细胞和 CD3 T 淋巴细胞的增殖。二价疫苗候选物和 FKC 组的 TA、VA 和 PA 均显著高于对照组。CD3、IgM、CD4-1、CD4-2、CD8α和 CD8β 基因上调。鳗弧菌攻毒后，AF、KF、RF 和 FKC 组的 RPS 分别为 62.6±2.33%、78.95±3.01%、75.45±0.97%和 56.71±2.15%。结果表明，三种二价疫苗候选物和 FKC 均可诱导牙鲆产生免疫应答，对鳗弧菌具有良好的保护作用，KF 可作为一种有效的二价亚单位疫苗候选物。
