Laboratório de Desenvolvimento de Vacinas, Instituto Butantan, São Paulo, Brazil.
York Biomedical Research Institute, University of York, York, United Kingdom.
Front Immunol. 2021 Mar 3;11:624613. doi: 10.3389/fimmu.2020.624613. eCollection 2020.
The radiation-attenuated cercarial vaccine remains the gold standard for the induction of protective immunity against . Furthermore, the protection can be passively transferred to naïve recipient mice from multiply vaccinated donors, especially IFNgR KO mice. We have used such sera versus day 28 infection serum, to screen peptide arrays and identify likely epitopes that mediate the protection. The arrays encompassed 55 secreted or exposed proteins from the alimentary tract and tegument, the principal interfaces with the host bloodstream. The proteins were printed onto glass slides as overlapping 15mer peptides, reacted with primary and secondary antibodies, and reactive regions detected using an Agilent array scanner. Pep Slide Analyzer software provided a numerical value above background for each peptide from which an aggregate score could be derived for a putative epitope. The reactive regions of 26 proteins were mapped onto crystal structures using the CCP4 molecular graphics, to aid selection of peptides with the greatest accessibility and reactivity, prioritizing vaccine over infection serum. A further eight MEG proteins were mapped to regions conserved between family members. The result is a list of priority peptides from 44 proteins for further investigation in multiepitope vaccine constructs and as targets of monoclonal antibodies.
辐射减毒尾蚴疫苗仍然是诱导针对 的保护性免疫的金标准。此外，这种保护可以从多次接种的供体被动转移到幼稚的受体小鼠，特别是 IFNgR KO 小鼠。我们使用这种血清与第 28 天感染的血清相比，筛选肽阵列并确定可能介导保护的表位。该阵列包括来自消化道和表皮的 55 种分泌或暴露蛋白，是与宿主血液接触的主要界面。这些蛋白被打印到玻璃载玻片上作为重叠的 15 聚体肽，与一抗和二抗反应，并使用安捷伦阵列扫描仪检测反应区域。Pep Slide Analyzer 软件为每个肽提供了一个高于背景的数值，从中可以得出一个假定表位的总得分。使用 CCP4 分子图形将 26 种蛋白质的反应区域映射到晶体结构上，以帮助选择具有最大可及性和反应性的肽，优先选择疫苗而不是感染血清。另外 8 种 MEG 蛋白被映射到家族成员之间保守的区域。结果是一个来自 44 种蛋白质的优先肽列表，用于进一步研究多表位疫苗构建体，并作为单克隆抗体的靶标。
