Palenzuela Oswaldo, Sitjà-Bobadilla Ariadna, Riaza Ana, Silva Raquel, Arán Javier, Alvarez-Pellitero Pilar
Instituto de Acuicultura Torre de la Sal (CSIC), 12595 Ribera de Cabanes, Castell6n, Spain.
Dis Aquat Organ. 2009 Sep 23;86(2):123-34. doi: 10.3354/dao02102.
The kinetics of the antibody production and the protection at challenge were studied in turbot inoculated with various scuticociliate antigen preparations: live ciliates putatively attenuated through long-term in vitro culture (Trial 1) and formalin-killed ciliates without or with GERBU adjuvant in Trials 2, 3, and 4. Antigen used in killed preparations was a mixture of 3 different ciliate isolates (V3) in the case of Trials 2 and 3, whereas in Trial 4, monovalent (V1), trivalent (V3) or pentavalent (V5) antigens were used. A booster injection was administered 28 to 29 d post-priming in all trials. Fish were challenged with virulent live ciliates after the immunization protocol, testing 2 challenge times in Trial 2 (t1 and t2). No protection was obtained in Trial 1 with live ciliates, which in turn were not completely attenuated. Using killed-ciliate formulations, protection was high only in Trial 3 when a low dose (50 000 ciliates fish(-1)) was used for challenge. In Trial 1, heat-inactivated sera of antigen-inoculated fish agglutinated the homologous ciliate, although no specific antibodies were detectable by ELISA. In contrast, high specific antibody levels were detected in antigen-inoculated fish in Trials 2 and 4, and their amount increased progressively, usually peaking after challenge. No advantage was obtained from the use of V5 antigens compared to V1 or V3. No good correlation was observed in most cases between serum antibody levels and protection. Although the use of GERBU adjuvant generally increased the specific immune response, some undesired side effects indicate a need to adjust dosage and/or improve the formulation.
在试验1中使用经长期体外培养假定已减毒的活纤毛虫；在试验2、3和4中使用福尔马林灭活的纤毛虫，分别添加或不添加GERBU佐剂。在试验2和3中，灭活制剂中使用的抗原是3种不同纤毛虫分离株的混合物（V3），而在试验4中，使用的是单价（V1）、三价（V3）或五价（V5）抗原。在所有试验中，初次免疫后28至29天进行加强注射。免疫程序结束后，用强毒活纤毛虫对鱼进行攻毒，在试验2中测试了2个攻毒时间点（t1和t2）。试验1中使用活纤毛虫未获得保护，这些活纤毛虫并未完全减毒。使用灭活纤毛虫制剂时，仅在试验3中当使用低剂量（50 000个纤毛虫/鱼）进行攻毒时才有较高的保护率。在试验1中，接种抗原的鱼的热灭活血清能凝集同源纤毛虫，尽管通过ELISA检测不到特异性抗体。相比之下，在试验2和4中接种抗原的鱼中检测到高特异性抗体水平，并且其数量逐渐增加，通常在攻毒后达到峰值。与V1或V3相比，使用V5抗原没有优势。在大多数情况下，血清抗体水平与保护作用之间未观察到良好的相关性。尽管使用GERBU佐剂通常会增强特异性免疫反应，但一些不良副作用表明需要调整剂量和/或改进制剂。
