Alba Annia, Hernández Hilda M, Marcet Ricardo, Gil Alejandro L, Vázquez Antonio A, Figueredo Mabel, Sánchez Jorge, Garay Hilda E, Sarracent Jorge
Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, Havana, Cuba,
Parasitol Res. 2014 Sep;113(9):3185-93. doi: 10.1007/s00436-014-3981-y. Epub 2014 Jun 13.
The control of fasciolosis, as that of other vector-borne diseases, must be related to the control of the lymnaeid snails, the intermediate hosts of the parasite. Thus, an accurate epidemiological surveillance of the transmission foci where the infected mollusks occur is essential. For this purpose, immunoassays could be a useful tool. However, information regarding specific proteins of intramolluscan larvae and previous studies concerning monoclonal antibody generation against asexual stages of trematodes are scarce. Therefore, we explored the antigenic features of intramolluscan rediae of Fasciola hepatica to evaluate three antigenic preparations in order to use the most promising one for developing specific monoclonal antibodies. Mouse antiserum was generated against each antigen for assessing the polyclonal antibody response against the crude extract of rediae and the cross-reactivity against lymnaeids. The specific C-terminal of F. hepatica cytochrome c oxidase subunit I (first antigen), selected by in silico analyses, might not be the appropriate target for immunoassay detection of infected snails, due to its low representation in the total extract of rediae. The majoritarian mixture of low-molecular-weight proteins (<30 kDa) from the rediae homogenate (second antigen) revealed a significant cross-reactivity with lymnaeids. Evidence of the existence of mimetic immunogenic epitopes in this fraction of F. hepatica rediae was achieved. High immunogenicity of the crude extract of rediae (third antigen), mainly related to parasite's specific epitopes, was regarded. Therefore, the rediae homogenate is stated as the most promising antigen from those evaluated, for monoclonal antibody development with potentialities for detecting F. hepatica-infected snails.
与其他媒介传播疾病一样，肝片吸虫病的防控必须与作为该寄生虫中间宿主的椎实螺的控制相关。因此，对受感染软体动物出现的传播疫源地进行准确的流行病学监测至关重要。为此，免疫测定可能是一种有用的工具。然而，关于软体动物内幼虫的特定蛋白质的信息以及先前有关针对吸虫无性阶段产生单克隆抗体的研究很少。因此，我们探究了肝片吸虫软体动物内雷蚴的抗原特性，以评估三种抗原制剂，以便使用最有前景的一种来开发特异性单克隆抗体。针对每种抗原制备小鼠抗血清，以评估针对雷蚴粗提物的多克隆抗体反应以及对椎实螺的交叉反应性。通过计算机分析选择的肝片吸虫细胞色素c氧化酶亚基I的特定C末端（第一种抗原），由于其在雷蚴总提取物中的含量较低，可能不是免疫测定检测受感染蜗牛的合适靶标。雷蚴匀浆中低分子量蛋白质（<30 kDa）的主要混合物（第二种抗原）显示出与椎实螺有显著的交叉反应性。在肝片吸虫雷蚴的这一部分中发现了模拟免疫原性表位的证据。雷蚴粗提物（第三种抗原）具有高免疫原性，主要与寄生虫的特异性表位有关。因此，雷蚴匀浆被认为是所评估的最有前景的抗原，可用于开发具有检测肝片吸虫感染蜗牛潜力的单克隆抗体。
