The recombinant gut-associated M17 leucine aminopeptidase in combination with different adjuvants confers a high level of protection against Fasciola hepatica infection in sheep
The recombinant gut-associated M17 leucine aminopeptidase in combination with different adjuvants confers a high level of protection against Fasciola hepatica infection in sheep
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DOI：
10.1016/j.vaccine.2011.09.020
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发表时间：
2011-11-08
期刊：
影响因子：
5.5
通讯作者：
Carmona, Carlos
中科院分区：
文献类型：
--
作者：
Maggioli, Gabriela;Acosta, Daniel;Carmona, Carlos
肝片形吸虫 M17 亮氨酸氨肽酶 (FhLAP) 被认为在分解代谢由肠道相关内肽酶对宿主多肽的协同活性产生的肽中发挥作用，从而释放用于蛋白质合成代谢的氨基酸。在这项研究中，在 Corriedale 绵羊身上进行的疫苗接种试验中，将大肠杆菌中产生的这种同源六聚金属肽酶的重组功能形式与不同类型的佐剂结合使用，以对抗肝囊蚴菌的实验攻击。实验测定由 6 组，每组 10 只动物组成；其中5个组(1-5)在第0周和第4周皮下接种与弗氏完全加不完全佐剂(组1)、明矾(组2)混合的100μg rFhLAP。 Adyuvac 50（第 3 组）、DEAE-D（第 4 组）和 Ribi（第 5 组）；佐剂对照组（第6组）接受弗氏佐剂。加强免疫后两周，对绵羊进行 200 个囊蚴的口服攻击。 rFhLAP 免疫导致所有接种组的吸虫负担显着降低：与佐剂对照组相比，弗氏组为 83.8%，Alum 组为 86.7%，Adyuvac 50 组为 74.4%，Ribi 组为 49.8%，DEAE-D 组为 49.5%。对回收的肝吸虫的形态分析显示，不同疫苗接种组的大小没有显着变化。所有疫苗制剂均引发特异性 IgG、IgG1 和 IgG2 反应。这项研究表明，基于 rFhLAP 的肝吸虫疫苗与不同佐剂相结合，可显着降低反刍动物中的蠕虫负担，而在接受酶以及商业批准的佐剂 Alum 和 Adyuvac 50 的动物中，蠕虫负担较高。(C) 2011 Elsevier Ltd. 保留所有权利。
Fasciola hepatica M17 leucine aminopeptidase (FhLAP) is thought to play a role in catabolizing peptides generated by the concerted activity of gut-associated endopeptidases on host polypeptides, thus releasing amino acids to be used in protein anabolism. In this study, a recombinant functional form of this homo hexameric metallopeptidase produced in Escherichia coli was used in combination with adjuvants of different types in a vaccination trial in Corriedale sheep against experimental challenge with F. hepatica metacercariae. The experimental assay consisted of 6 groups of 10 animals; 5 of the groups (1-5) were subcutaneously inoculated at weeks 0 and 4 with100 mu g of rFhLAP mixed with Freund's complete plus incomplete adjuvant (group 1), Alum (group 2). Adyuvac 50 (group 3), DEAE-D (group 4) and Ribi (group 5); the adjuvant control group (group 6) received Freund's adjuvant. Two weeks after the booster, the sheep were orally challenged with 200 metacercariae. Immunization with rFhLAP induced significant reduction in fluke burdens in all vaccinated groups: 83.8% in the Freund's group, 86.7% in the Alum group, 74.4% in the Adyuvac 50 group, 49.8% in the Ribi group and 49.5% in the DEAE-D group compared to the adjuvant control group. Morphometric analysis of recovered liver flukes showed no significant size modifications in the different vaccination groups. All vaccine preparations elicited specific IgG, IgG1 and IgG2 responses. This study shows that a liver fluke vaccine based on rFhLAP combined with different adjuvants significantly reduced worm burden in a ruminant species that was high in animals that received the enzyme along with the commercially approved adjuvants Alum and Adyuvac 50. (C) 2011 Elsevier Ltd. All rights reserved.