Bień Justyna, Sałamatin Rusłan, Sulima Anna, Savijoki Kirsi, Bruce Conn David, Näreaho Anu, Młocicki Daniel
Acta Parasitol. 2016 Mar;61(2):429-42. doi: 10.1515/ap-2016-0058.
Hymenolepis diminuta is an important model species in studies of therapeutics, biochemical processes, immune responses and other aspects of cestodiasis. The parasite produces numerous excretory-secretory (E-S) proteins and a glycocalyx covering its body. Our study focused on the mass spectrometry analysis of the E-S material with an objective to determine if E-S contains any new proteins, in particular those that can be identified as: antigens, vaccine candidates and drug targets. These proteins might engage directly in host-parasite interactions. Adult parasites collected from experimentally infected rats were cultured in vitro for 5 and 18h. Immunoblotting was used to verify which E-S protein bands separated in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) react with specific antibodies from sera of infected rats. We identified thirty-nine proteins by LC-MS/MS (liquid chromatography mass spectrometry). Results indicated the presence of proteins that have never been identified in cestode E-S material. Immunoblotting showed the immunogenicity of E-S products of H. diminuta, most probably associated with the presence of proteins known as antigens in other flatworm species. Among identified proteins are those engaged in immunomodulatory processes (eg. HSP), in response to oxidative stress (peroxidasin) or metabolism (eg. GAPDH). The predominant functions are associated with metabolism and catalytic activity. This is the first study identifying E-S-proteins in adult tapeworms, thus providing information for better understanding host-parasite interrelationships, and may point out potential targets for vaccines or drug discovery studies, as among the proteins observed in our study are those known to be antigens.
微小膜壳绦虫是治疗学、生化过程、免疫反应及绦虫病其他方面研究中的重要模式物种。该寄生虫会产生大量排泄-分泌(E-S)蛋白以及覆盖其身体的糖萼。我们的研究聚焦于对E-S物质进行质谱分析,目的是确定E-S中是否含有任何新蛋白质,特别是那些可被鉴定为抗原、候选疫苗和药物靶点的蛋白质。这些蛋白质可能直接参与宿主与寄生虫的相互作用。从实验感染大鼠体内收集的成虫在体外培养5小时和18小时。采用免疫印迹法来验证在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)中分离出的哪些E-S蛋白条带能与感染大鼠血清中的特异性抗体发生反应。我们通过液相色谱-质谱联用(LC-MS/MS)鉴定出了39种蛋白质。结果表明存在从未在绦虫E-S物质中鉴定出的蛋白质。免疫印迹显示微小膜壳绦虫E-S产物具有免疫原性,很可能与其他扁虫物种中被称为抗原的蛋白质的存在有关。在鉴定出的蛋白质中,有参与免疫调节过程的(如热休克蛋白)、应对氧化应激的(过氧化物酶)或参与代谢的(如甘油醛-3-磷酸脱氢酶)。主要功能与代谢和催化活性相关。这是首次对成虫绦虫的E-S蛋白进行鉴定的研究,从而为更好地理解宿主-寄生虫相互关系提供了信息,并且可能指出疫苗或药物研发研究的潜在靶点,因为在我们的研究中观察到的蛋白质中有已知的抗原。