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.
Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, PR China.
Mol Immunol. 2018 Dec;104:79-89. doi: 10.1016/j.molimm.2018.11.001. Epub 2018 Nov 14.
CD40 is a crucial signal mediating factor in T-dependent B cell responses and involved in many aspects of cellular and humoral immunity. In this paper, recombinant protein of CD40 in flounder (Paralichthys olivaceus) and its antibodies (Abs) were produced, native CD40 molecules in flounder tissues were identified, then the CD40 leukocytes in T/B lymphocytes were characterized, and the variations of CD40 leukocytes in flounder after Hirame novirhabdovirus (HIRRV) infection and immunization were investigated, respectively. The results showed that the Abs could specifically recognize native flounder CD40 molecule at 32 kDa. The proportions of CD40 leukocytes were varied by flounder tissues. CD40/IgM B lymphocytes, CD40/CD4-1 T lymphocytes, CD40/CD4-2 T lymphocytes and CD40/CD8 T lymphocytes in peripheral blood leukocytes (PBLs) were 1.18 ± 0.27%, 0.69 ± 0.17%, 0.75 ± 0.14% and 0.25 ± 0.14%; were 2.80 ± 0.32%, 0.71 ± 0.19%, 0.88 ± 0.23% and 0.33 ± 0.17% in spleen; 4.11 ± 0.47%, 0.92 ± 0.18%, 1.09 ± 0.17% and 0.9 ± 0.17% in head kidney; 1.92 ± 0.39%, 1.02 ± 0.23%, 1.33 ± 0.38% and 0.67 ± 0.24% in intestine; 1.24 ± 0.36%, 1.21 ± 0.24%, 1.70 ± 0.3% and 0.97 ± 0.21% in gill, respectively. The percentages of CD40 leukocytes in PBLs were significantly increased in both HIRRV infection and immunization groups, and reached their peak levels at 3rd day with 5.70 ± 0.16% and 6.40 ± 0.13%, respectively. Concluded with our previous study, these data first reported that CD40 molecules were expressed on both B and T lymphocytes in teleost, and had a coordination with T and B lymphocytes in immune responses.
CD40 是 T 细胞依赖的 B 细胞反应中重要的信号介导因子，参与细胞和体液免疫的多个方面。本文制备了牙鲆（Paralichthys olivaceus）CD40 重组蛋白及其抗体（Abs），鉴定了牙鲆组织中的天然 CD40 分子，然后对 T/B 淋巴细胞中的 CD40 白细胞进行了特征分析，并分别研究了牙鲆感染虹彩病毒（HIRRV）和免疫后的 CD40 白细胞变化。结果表明，Abs 可特异性识别 32kDa 的天然牙鲆 CD40 分子。CD40 白细胞在牙鲆组织中的比例不同。外周血白细胞（PBLs）中 CD40/IgM B 淋巴细胞、CD40/CD4-1 T 淋巴细胞、CD40/CD4-2 T 淋巴细胞和 CD40/CD8 T 淋巴细胞分别为 1.18±0.27%、0.69±0.17%、0.75±0.14%和 0.25±0.14%；在脾脏中分别为 2.80±0.32%、0.71±0.19%、0.88±0.23%和 0.33±0.17%；在头肾中分别为 4.11±0.47%、0.92±0.18%、1.09±0.17%和 0.90±0.17%；在肠中分别为 1.92±0.39%、1.02±0.23%、1.33±0.38%和 0.67±0.24%；在鳃中分别为 1.24±0.36%、1.21±0.24%、1.70±0.30%和 0.97±0.21%。HIRRV 感染和免疫组的 PBLs 中 CD40 白细胞的百分比均显著增加，在第 3 天分别达到 5.70±0.16%和 6.40±0.13%的峰值水平。结合我们之前的研究，这些数据首次报道了 CD40 分子在硬骨鱼的 B 和 T 淋巴细胞上表达，并在免疫反应中与 T 和 B 淋巴细胞协同作用。
