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含有来自莫氏钝缘蜱保护性表位的新型唾液抗止血剂:其单独及联合疫苗效力评估

New salivary anti-haemostatics containing protective epitopes from Ornithodoros moubata ticks: Assessment of their individual and combined vaccine efficacy.

作者信息

Díaz-Martín Verónica, Manzano-Román Raúl, Oleaga Ana, Pérez-Sánchez Ricardo

机构信息

Parasitología Animal, Instituto de Recursos Naturales y Agrobiología de Salamanca (IRNASA, CSIC), Cordel de Merinas, 40-52, 37008 Salamanca, Spain.

出版信息

Vet Parasitol. 2015 Sep 15;212(3-4):336-49. doi: 10.1016/j.vetpar.2015.08.005. Epub 2015 Aug 7.

Abstract

Ornithodoros moubata is the main vector of the pathogens causing African swine fever and human relapsing fever in Africa. The development of an efficient vaccine against this tick would facilitate its control and the prevention of the diseases it transmits to a considerable extent. Previous efforts to identify vaccine target candidates led us to the discovery of novel salivary proteins that probably act as anti-haemostatics at the host-tick interface, including a secreted phospholipase A2 (PLA2), a 7DB-like protein (7DB-like), a riboprotein 60S L10 (RP-60S), an apyrase (APY), and a new platelet aggregation inhibitor peptide, designated mougrin (MOU). In this work, the corresponding recombinant proteins were expressed in Escherichia coli and their individual vaccine efficacy was tested in rabbit vaccination trials. All of them, except the less immunogenic RP-60S, induced strong humoral responses that reduced tick feeding and survival, providing vaccine efficacies of 44.2%, 43.2% and 27.2%, 19.9% and 17.3% for PLA2, APY, MOU, RP-60S and 7DB-like, respectively. In the case of the more protective recombinant antigens (PLA2, APY and MOU), the immunodominant protective linear B-cell epitopes were identified and their combined vaccine efficacy was tested in a second vaccine trial using different adjuvants. In comparison with the best efficacy of individual antigens, the multicomponent vaccine increased vaccine efficacy by 13.6%, indicating additive protective effects rather than a synergistic effect. Tick saliva inoculated during natural tick-host contacts had a boosting effect on vaccinated animals, increasing specific antibody levels and protection.

摘要

钝缘蜱是非洲引起非洲猪瘟和人类回归热的病原体的主要传播媒介。研发一种针对这种蜱虫的高效疫苗将在很大程度上有助于对其进行控制并预防它所传播的疾病。此前鉴定疫苗候选靶点的工作使我们发现了一些新的唾液蛋白,这些蛋白可能在宿主与蜱虫的界面处起到抗止血作用,包括一种分泌型磷脂酶A2(PLA2)、一种类7DB蛋白(7DB-like)、一种核糖体蛋白60S L10(RP - 60S)、一种腺苷三磷酸双磷酸酶(APY)以及一种新的血小板聚集抑制肽,命名为mougrin(MOU)。在这项研究中,相应的重组蛋白在大肠杆菌中表达,并在兔疫苗接种试验中测试了它们各自的疫苗效力。除免疫原性较弱的RP - 60S外,所有其他蛋白均诱导了强烈的体液免疫反应,减少了蜱虫的取食和存活,PLA2、APY、MOU、RP - 60S和7DB-like的疫苗效力分别为44.2%、43.2%、27.2%、19.9%和17.3%。对于保护性更强的重组抗原(PLA2、APY和MOU),鉴定了免疫显性的保护性线性B细胞表位,并在第二次疫苗试验中使用不同佐剂测试了它们联合的疫苗效力。与单个抗原的最佳效力相比,多组分疫苗使疫苗效力提高了13.6%,表明是相加的保护作用而非协同作用。在蜱虫与宿主自然接触期间接种的蜱虫唾液对已接种疫苗的动物有增强作用,提高了特异性抗体水平和保护效果。

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