Jing Xing, Wenbin Zhan
Laboratory of Pathology and Immunology of Aquatic Animals, LMMEC, Ocean University of China, Qingdao 266003, PR China.
Fish Shellfish Immunol. 2006 Apr;20(4):528-35. doi: 10.1016/j.fsi.2005.07.001. Epub 2005 Sep 12.
In this paper, haemocyte antigenicity of seven bivalve species (scallop (Chlamys farreri), bay scallop (Argoecten irradians), oyster (Crassostrea talienwhanensis), asiatic hard clam (Meretrix meretrix), monila clam (Ruditapes philipinarum), purplish washington clam (Saxidomus purpuratus) and horny ark (Scapharca subcrenta)) were analysed using monoclonal antibodies (MAbs) 1E7, 1F12, 2C6 and 2H5 against haemocytes of C. farreri, employed methods of immuno-dotblotting (IDB), indirect immunofluorescence assay (IIFA) and western-blotting (WB). The four MAbs react with haemocytes of seven bivalve species. As the results for both IDB and IIFA, MAb 1E7 was positive with haemocytes of R. philipinarum, MAb 1F12 with haemocytes of A. irradians, M. meretrix, R. philipinarum and S. purpuratus; MAb 2C6 with haemocytes of the other five bivalve species except for S. purpuratus. MAb 2H5 was negative with haemocytes of the other six bivalve species in IDB, but was positive with haemocytes of R. philipinarum and S. purpuratus in IIFA. Further experiments by WB showed MAb 1F12 was able to recognise the protein of A. irradians haemocyte at molecular weights of 156 and 80 kDa, haemocytes of M. meretris, R. philipinarum, S. purpuratus, at 60, 30, 58 kDa, respectively. MAb 2C6 recognised haemocyte M. meretris proteins at 50 and 37 kDa, A. irradians, C. talienwhanensis, R. philipinarum, S. subcrenta at 40, 38, 38, 45 kDa, respectively. There were no protein bands reacting with MAb 1E7 and MAb 2H5. The results indicate antigenic similarities exist among haemocytes of the seven bivalve species.
本文利用抗栉孔扇贝血细胞的单克隆抗体（MAb）1E7、1F12、2C6和2H5，采用免疫斑点印迹（IDB）、间接免疫荧光分析（IIFA）和蛋白质印迹（WB）方法，分析了7种双壳贝类（栉孔扇贝（Chlamys farreri）、海湾扇贝（Argoecten irradians）、近江牡蛎（Crassostrea talienwhanensis）、文蛤（Meretrix meretrix）、菲律宾蛤仔（Ruditapes philipinarum）、紫石房蛤（Saxidomus purpuratus）和毛蚶（Scapharca subcrenta））的血细胞抗原性。这4种单克隆抗体与7种双壳贝类的血细胞发生反应。IDB和IIFA结果显示，MAb 1E7与菲律宾蛤仔的血细胞呈阳性反应，MAb 1F12与海湾扇贝、文蛤、菲律宾蛤仔和紫石房蛤的血细胞呈阳性反应；MAb 2C6与除紫石房蛤外的其他5种双壳贝类的血细胞呈阳性反应。IDB中MAb 2H5与其他6种双壳贝类的血细胞呈阴性反应，但在IIFA中与菲律宾蛤仔和紫石房蛤的血细胞呈阳性反应。WB进一步实验表明，MAb 1F12能够识别分子量为156 kDa和80 kDa的海湾扇贝血细胞蛋白，文蛤、菲律宾蛤仔、紫石房蛤的血细胞蛋白分子量分别为60 kDa、30 kDa、58 kDa。MAb 2C6识别文蛤血细胞蛋白分子量为50 kDa和37 kDa，海湾扇贝、近江牡蛎、菲律宾蛤仔、毛蚶的血细胞蛋白分子量分别为40 kDa、38 kDa、38 kDa、45 kDa。没有与MAb 1E7和MAb 2H5反应的蛋白条带。结果表明7种双壳贝类的血细胞之间存在抗原相似性。
