Cobb C S, Levy M G, Noga E J
Department of Microbiology, Pathology, and Parasitology, North Carolina State University College of Veterinary Medicine, Raleigh 27606, USA.
Dis Aquat Organ. 1998 Oct 8;34(2):125-33. doi: 10.3354/dao034125.
The dinoflagellate Amyloodinium ocellatum, which causes amyloodiniosis or 'marine velvet disease', is one of the most serious ectoparasitic diseases plaguing warmwater marine fish culture worldwide. We report that tomato clownfish Amphiprion frenatus develop strong immunity to Amyloodinium ocellatum infection following repeated nonlethal challenges and that specific antibodies are associated with this response. Reaction of immune fish antisera against dinospore and trophont-derived antigens in Western blots indicated both shared and stage-specific antibody-antigen reactions. A mannan-binding-protein affinity column was used to isolate IgM-like antibody from A. frenatus serum. The reduced Ig consisted of one 70 kD heavy chain and one 32 kD light chain with an estimated molecular weight of 816 kD for the native molecule. Immunoglobulin (Ig) isolated from immune but not non-immune fish serum significantly inhibited parasite infectivity in vitro. An enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal rabbit antibody produced against affinity-purified A. frenatus Ig. Anti-Amyloodinium serum antibody was not always detectable in immune fish, although serum antibody titers in immune fish increased after repeated exposure to the parasite. These results suggest that there may be a localized antibody response in skin/gill epithelial tissue, although antibody was rarely detected in skin mucus.
可导致淀粉卵甲藻病或“海水天鹅绒病”的双鞭毛藻眼点淀粉卵甲藻，是困扰全球温水海水鱼类养殖的最严重的体外寄生虫病之一。我们报告说，番茄小丑鱼（双带双锯鱼）在反复接受非致死性攻击后，对眼点淀粉卵甲藻感染产生了强大的免疫力，并且特定抗体与这种反应相关。免疫鱼抗血清在蛋白质免疫印迹中对动孢子和营养体衍生抗原的反应表明存在共同的和阶段特异性的抗体-抗原反应。使用甘露聚糖结合蛋白亲和柱从双带双锯鱼血清中分离出类IgM抗体。还原后的Ig由一条70 kD重链和一条32 kD轻链组成，天然分子的估计分子量为816 kD。从免疫而非非免疫鱼血清中分离出的免疫球蛋白（Ig）在体外显著抑制了寄生虫的感染性。利用针对亲和纯化的双带双锯鱼Ig产生的多克隆兔抗体开发了一种酶联免疫吸附测定（ELISA）。尽管免疫鱼在反复接触寄生虫后血清抗体滴度有所增加，但在免疫鱼中并非总能检测到抗眼点淀粉卵甲藻血清抗体。这些结果表明，尽管在皮肤黏液中很少检测到抗体，但在皮肤/鳃上皮组织中可能存在局部抗体反应。
