Drennan John D, Lapatra Scott E, Swan Christine M, Ireland Sue, Cain Kenneth D
Department of Fish and Wildlife Resources and the Aquaculture Research Institute, University of Idaho, 6th and Line Street, CRN Building, Moscow, ID 83844-1136, USA.
Fish Shellfish Immunol. 2007 Sep;23(3):657-69. doi: 10.1016/j.fsi.2007.01.016. Epub 2007 Feb 1.
Serum and cutaneous mucus antibodies were monitored in white sturgeon for 15 weeks following intraperitoneal immunization. Ten fish were immunized (50 microg) with white sturgeon iridovirus (WSIV) or white sturgeon gonad (WSGO) tissue culture cells emulsified with or without FCA. An additional group was immunized with FITC:KLH+FCA. Fish were booster immunized at 6 weeks. Fish immunized with FITC:KLH+FCA produced significant serum antibodies to FITC by 6 weeks and this response peaked at 12 weeks (average titer 31,000). Mucosal antibodies to FITC were first detected at 12 weeks and significantly elevated by 15 weeks (average titer 18). Anti-WSIV antibody titers were detected in the serum by 9 weeks in fish immunized with WSIV and WSIV+FCA, but only a small number responded to immunization. At 15 weeks, four fish immunized with WSIV produced serum antibodies (average titer 838) and one fish immunized with WSIV+FCA had a serum titer of 1600. Mucosal anti-WSIV antibody titers of 8 and 16 were observed in two fish from the WSIV group at 12 weeks while four different fish from this group responded at 15 weeks (average titer 4). Western Blot using a monoclonal antibody confirmed immunoglobulin in mucus, and specificity to WSIV was further demonstrated by immunocytochemistry using serum from fish immunized with WSIV. Specific antibody was not detected in mucus of fish immunized with WSIV+FCA, WSGO, or WSGO+FCA. Collectively, these experiments demonstrate that white sturgeon can generate a specific antibody response following immunization, and is the first report showing mucosal immunoglobulin is present in this species.
在腹腔免疫后,对白鲟的血清和皮肤黏液抗体进行了15周的监测。用白鲟虹彩病毒(WSIV)或白鲟性腺(WSGO)组织培养细胞乳化后(添加或不添加弗氏完全佐剂(FCA))对10条鱼进行免疫(50微克)。另一组用异硫氰酸荧光素:钥孔戚血蓝蛋白+弗氏完全佐剂(FITC:KLH+FCA)进行免疫。在第6周对鱼进行加强免疫。用FITC:KLH+FCA免疫的鱼在6周时产生了针对FITC的显著血清抗体,这种反应在12周时达到峰值(平均滴度31,000)。针对FITC的黏膜抗体在12周时首次检测到,并在15周时显著升高(平均滴度18)。在用WSIV和WSIV+FCA免疫的鱼中,在9周时血清中检测到抗WSIV抗体滴度,但只有少数鱼对免疫有反应。在15周时,4条用WSIV免疫的鱼产生了血清抗体(平均滴度838),1条用WSIV+FCA免疫的鱼血清滴度为1600。在12周时,WSIV组的两条鱼的黏膜抗WSIV抗体滴度分别为8和16,而该组的另外4条不同的鱼在15周时有反应(平均滴度4)。使用单克隆抗体的蛋白质免疫印迹法证实了黏液中的免疫球蛋白,并用来自WSIV免疫鱼的血清进行免疫细胞化学进一步证明了对WSIV的特异性。在用WSIV+FCA、WSGO或WSGO+FCA免疫的鱼的黏液中未检测到特异性抗体。总体而言,这些实验表明白鲟在免疫后可以产生特异性抗体反应,这是首次报道该物种中存在黏膜免疫球蛋白。