Palaksha K J, Shin Gee-Wook, Kim Young-Rim, Jung Tae-Sung
Laboratory of Fish & Shellfish Diseases, College of Veterinary Medicine, Gyeongsang National University, Jinju, 660-701, South Korea.
Fish Shellfish Immunol. 2008 Apr;24(4):479-88. doi: 10.1016/j.fsi.2008.01.005. Epub 2008 Jan 11.
The innate immune system, particularly the external body surface, plays a frontier role in protecting fish under intensive aquaculture and at prolonged low temperatures from relevant infections due to inadequate adaptive immune responses. In the present study we aimed to understand the mucosal immunity of an economically important mariculture fish, olive flounder (Paralichthys olivaceus) by evaluating the immune components from its skin mucus. The activities of lysozyme (233.33+/-171.82 units mg(-1)), trypsin-like protease (42.84+/-1.249 units mg(-1)), alkaline phosphatase (0.376+/-0.005 units mg(-1)) and esterase (0.170+/-0.006 units mg(-1)) were detected in the skin mucus. Transferrin was identified by MALDI-TOF/MS analysis. ELISA and immunoblot assays using anti-flounder IgM monoclonal antibody showed the presence of a significant level (1.80+/-0.001, n=3) of monomer immunoglobulin M (IgM) with approximate molecular weight of 160 and 25 kDa under non-denaturing and denaturing states, respectively. Skin mucus showed strong antibacterial activity against tested fish pathogenic bacteria. In addition, skin mucus successfully agglutinated (HA titre 2(8)), but completely failed to haemolyse, rabbit erythrocytes. In conclusion, the major immune components of the skin mucus, identified in the present study, are possibly involved in the broad spectrum non-specific immunity of olive flounder.
在集约化水产养殖和长期低温环境下，由于适应性免疫反应不足，鱼类易受相关感染，而先天免疫系统，尤其是鱼体体表，在保护鱼类方面发挥着前沿作用。在本研究中，我们旨在通过评估经济价值重要的海水养殖鱼类牙鲆（Paralichthys olivaceus）皮肤黏液中的免疫成分，来了解其黏膜免疫。在牙鲆皮肤黏液中检测到了溶菌酶（233.33±171.82单位mg(-1)）、类胰蛋白酶（42.84±1.249单位mg(-1)）、碱性磷酸酶（0.376±0.005单位mg(-1)）和酯酶（0.170±0.006单位mg(-1)）的活性。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF/MS）分析鉴定出了转铁蛋白。使用抗牙鲆IgM单克隆抗体进行的酶联免疫吸附测定（ELISA）和免疫印迹分析表明，在非变性和变性状态下，分别存在显著水平（1.80±0.001，n = 3）的单体免疫球蛋白M（IgM），其近似分子量分别为160 kDa和25 kDa。牙鲆皮肤黏液对受试鱼类病原菌表现出较强的抗菌活性。此外，牙鲆皮肤黏液成功凝集（血凝效价2(8)），但完全不能溶解兔红细胞。总之，本研究中鉴定出的牙鲆皮肤黏液主要免疫成分可能参与了牙鲆的广谱非特异性免疫。
