National Bureau of Fish Genetic Resources, Fish Health Management Division, Canal Ring Road, P.O. Dilkusha, Lucknow 226002, Uttar Pradesh, India.
Fish Shellfish Immunol. 2011 Feb;30(2):569-75. doi: 10.1016/j.fsi.2010.12.004. Epub 2010 Dec 15.
Snakehead Channa striata is an important freshwater food fish in many Southeast Asian countries. Three monoclonal antibodies (C9, C10 and D10) were developed against purified serum immunoglobulins of Channa striata (Cs-Ig) and characterized. C9 and D10 MAbs were specific to heavy chain, while C10 MAb detected only unreduced Cs-Ig in western blotting. In competitive ELISA, C9 and C10 MAbs were specific to C. striata Ig and showed no cross reactivity with serum Ig of other fish species i.e. Channa punctatus, Channa marulius, Clarias batrachus and Labeo rohita. D10 MAb showed reactivity to serum Ig of C. striata and C. marulius. In FACS analysis of gated lymphocytes, the percentage of Ig+ cells detected by C9 MAb was 18.2%, 27.7% and 10.3% in blood, spleen and kidney, respectively (n=3, body weight 500-600 g). However, only a few cells (0.5%) were found to be Ig+ in thymus (n=5). C9 MAb was also successfully employed to demonstrate Ig+ cells in blood smears and formalin fixed sections of spleen and kidney. These findings suggest that the spleen plays an important role in humoral immunity as compared to head kidney. Further, these MAbs can be useful immunological tool in monitoring health status of cultured C. striata.
条纹攀鲈是东南亚国家重要的淡水食用鱼类。本研究制备了 3 株针对纯化的条纹攀鲈血清免疫球蛋白(Cs-Ig)的单克隆抗体(C9、C10 和 D10),并对其进行了鉴定。C9 和 D10 MAb 特异性识别重链,而 C10 MAb 在 Western blot 中仅检测到未还原的 Cs-Ig。在竞争性 ELISA 中,C9 和 C10 MAb 特异性识别 C. striata Ig,与其他鱼类血清 Ig 无交叉反应,即斑鳢、乌鳢、革胡子鲶和罗非鱼。D10 MAb 对条纹攀鲈和乌鳢的血清 Ig 有反应性。在流式细胞术分析中,C9 MAb 检测到的 Ig+细胞百分比分别为血液、脾脏和肾脏中的 18.2%、27.7%和 10.3%(n=3,体重 500-600 g)。然而,在胸腺中仅发现少数细胞(0.5%)为 Ig+(n=5)。C9 MAb 还成功地用于显示血液涂片和脾脏及肾脏福尔马林固定切片中的 Ig+细胞。这些发现表明,与头肾相比,脾脏在体液免疫中发挥着重要作用。此外,这些单克隆抗体可作为监测养殖条纹攀鲈健康状况的有用免疫工具。