Yang Yong, Lacour Sandrine A, Lainé-Prade Véronique, Versillé Nicolas, Grasset-Chevillot Aurélie, Feng Shuang, Liu Ming Yuan, Boireau Pascal, Vallée Isabelle
Animal Health Laboratory,Université Paris-Est,Anses,ENVA,JRU BIPAR,Maisons-Alfort,France.
Key Laboratory of Zoonosis Research,Ministry of Education,Institute of Zoonosis,Jilin University,Changchun,People's Republic of China.
Parasitology. 2015 May;142(6):783-90. doi: 10.1017/S0031182014001851. Epub 2015 Jan 19.
Trichinella spiralis is an intracellular parasitic nematode of mammalian skeletal muscle, causing a serious zoonotic disease in humans and showing a high economic impact mainly in pig breeding. Serine proteinases of T. spiralis play important roles in the host-parasite interactions mediating host invasion. In this study, we have focused on newborn larvae (NBL-1), the first identified serine proteinase from the NBL stage of T. spiralis. Five monoclonal antibodies (mAbs) directed against the C-terminal part of NBL1, were produced. These mAbs were IgG1κ isotype and specifically recognized as a common motif of 10 amino acids (PSSGSRPTYP). Selected mAbs were further characterized using antigens from various developmental stages of T. spiralis. Western blot revealed that selected mAbs reacted with the native NBL1 at Mr 50 kDa in the adult and NBL mixed antigens and NBL stage alone. Indirect immunofluorescence analysis revealed that selected mAbs intensely stained only the embryos within the gravid females and the NBL. Thus, the produced mAbs are useful tools for the characterization of NBL1 as a major antigen of Trichinella involved in the invasion of the host but also for the development of new serological tests with an early detection of T. spiralis infection.
旋毛虫是一种寄生于哺乳动物骨骼肌细胞内的线虫，可导致人类严重的人畜共患病，主要对养猪业造成巨大经济影响。旋毛虫的丝氨酸蛋白酶在介导宿主入侵的宿主 - 寄生虫相互作用中发挥重要作用。在本研究中，我们聚焦于新生幼虫（NBL - 1），它是旋毛虫NBL阶段首次鉴定出的丝氨酸蛋白酶。我们制备了五种针对NBL1 C末端部分的单克隆抗体（mAb）。这些mAb为IgG1κ亚型，特异性识别一个由10个氨基酸组成的共有基序（PSSGSRPTYP）。使用旋毛虫不同发育阶段的抗原对筛选出的mAb进行进一步表征。蛋白质免疫印迹显示，筛选出的mAb与成虫和NBL混合抗原以及单独的NBL阶段中50 kDa的天然NBL1发生反应。间接免疫荧光分析显示，筛选出的mAb仅强烈染色孕虫体内的胚胎和NBL。因此，所产生的mAb是用于将NBL1鉴定为旋毛虫参与宿主入侵的主要抗原的有用工具，也是用于开发能够早期检测旋毛虫感染的新型血清学检测方法的有用工具。
