Sansri Veerawat, Meemon Krai, Changklungmoa Narin, Kueakhai Pornanan, Chantree Pathanin, Chaichanasak Pannigan, Lorsuwannarat Natcha, Itagaki Tadashi, Sobhon Prasert
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand; Faculty of Physical Therapy, Mahidol University, Phuttamonthon 4 Road, Salaya 73170, Nakhon Pathom, Thailand.
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
Vaccine. 2015 Mar 24;33(13):1596-601. doi: 10.1016/j.vaccine.2015.02.010. Epub 2015 Feb 17.
Fasciola gigantica cathepsin L1H (FgCatL1H) is one of the major cathepsin L released by juveniles of F. gigantica to aid in the invasion of host's tissues. Due to its high sequence similarity with other cathepsin L (CatL) isoforms of late stage F. gigantica, it was considered to be a good vaccine candidate that can block all CatL-mediated protease activities and affect juveniles as well as adult parasites. In this study, recombinant proFgCatL1H protein expressed in yeast, Pichia pastoris, system was mixed with Freund's adjuvants and used to subcutaneously immunize mice that were later challenged with metacercariae of F. gigantica. The percentage of worm protection in the rproFgCatL1H-vaccinated mice compared to the non-immunized and adjuvant control mice were approximately 62.7% and 66.1%, respectively. Anti-rproFgCatL1H antisera collected from vaccinated mice reacted specifically with rproFgCatL1H and other cathepsin L isoforms of F. gigantica, but the antibodies did not cross react with antigens from other trematode and nematode parasites, including Eurytrema pancreaticum, Opisthorchis viverrini, Fischoederius cobboldi, Cotylophoron cotylophorum, Gigantocotyle explanatum, Paramphistomum cervi, and Setaria labiato-papillosa. The levels of IgG1 and IgG2a in mouse sera increased significantly at two weeks after immunization and were highest during the sixth to eighth weeks after immunization. The IgG1 level was higher than IgG2a at all periods of immunization, implicating the dominance of the Th2 response. The levels of IgG1 and IgG2a in the immune sera were shown to be strongly correlated with the numbers of worm recovery, and the correlation coefficient was higher for IgG1. The levels of serum aspartate aminotransferase and alanine transaminase were significantly lower in the sera of rproFgCatL1H-vaccinated mice than in the infected control mice indicating a lower degree of liver damage. This study demonstrated a high potential of FgCatL1H vaccine, and its efficacy is currently being studied in the larger economic animals.
巨片形吸虫组织蛋白酶L1H(FgCatL1H)是巨片形吸虫幼虫释放的主要组织蛋白酶L之一,有助于其侵入宿主组织。由于它与巨片形吸虫后期的其他组织蛋白酶L(CatL)亚型具有高度的序列相似性,它被认为是一种很好的疫苗候选物,能够阻断所有CatL介导的蛋白酶活性,并对幼虫以及成虫期寄生虫产生影响。在本研究中,将在毕赤酵母系统中表达的重组前体FgCatL1H蛋白与弗氏佐剂混合,用于皮下免疫小鼠,随后用巨片形吸虫囊蚴对小鼠进行攻击。与未免疫和佐剂对照小鼠相比,接种rproFgCatL1H疫苗的小鼠的驱虫保护率分别约为62.7%和66.1%。从接种疫苗的小鼠收集的抗rproFgCatL1H抗血清与rproFgCatL1H以及巨片形吸虫的其他组织蛋白酶L亚型发生特异性反应,但这些抗体与其他吸虫和线虫寄生虫的抗原不发生交叉反应,包括胰阔盘吸虫、 viverrini 后睾吸虫、Fischoederius cobboldi、Cotylophoron cotylophorum、Gigantocotyle explanatum、鹿同盘吸虫和唇乳突丝虫。免疫后两周小鼠血清中的IgG1和IgG2a水平显著升高,在免疫后第六至八周达到最高。在整个免疫期间,IgG1水平高于IgG2a,这表明Th2反应占主导。免疫血清中IgG1和IgG2a的水平与虫体回收率密切相关,且IgG1的相关系数更高。接种rproFgCatL1H疫苗的小鼠血清中的天冬氨酸转氨酶和丙氨酸转氨酶水平显著低于感染对照小鼠,表明肝损伤程度较低。本研究证明了FgCatL1H疫苗具有很高的潜力,目前正在对更大的经济动物研究其疗效。