Basic Pathology Department, Federal University of Paraná, Curitiba, PR, Brazil.
Department of Bioprocess Engineering and Biotechnology, Federal University of Paraná, Curitiba, PR, Brazil.
J Immunol Res. 2017;2017:5871043. doi: 10.1155/2017/5871043. Epub 2017 Mar 7.
This work's goal was to research new candidate antigens for cutaneous leishmaniosis (CL). In order to reach the goal, we used random peptide phage display libraries screened using antibodies from patients. After selection, three peptides (P1, P2, and P3) were synthesized using Fmoc chemistry. The peptides individually or a mixture of them (MIX) was subsequently emulsified in complete and incomplete Freund's adjuvant and injected subcutaneously in golden hamsters. Sera from the hamsters administered with P1 presented antibodies that recognized proteins between 76 and 150 kDa from . Sera from hamsters which had peptides P2 and P3, as well as the MIX, administered presented antibodies that recognized proteins between 52 and 76 kDa of . The research on the similarity of the peptides' sequences in protein databases showed that they match a 63 kDa glycoprotein. The three peptides and the MIX were recognized by the sera from CL patients by immunoassay approach (ELISA). The peptides' MIX showed the best performance (79% sensitivity) followed by the P1 (72% sensitivity), and the AS presented 91% sensitivity. These results show a new route for discovering molecules for diagnosis or for immunoprotection against leishmaniosis.
这项工作的目的是研究皮肤利什曼病(CL)的新候选抗原。为了达到这个目标,我们使用针对患者的抗体筛选了随机肽噬菌体展示文库。经过选择,使用 Fmoc 化学合成了三个肽(P1、P2 和 P3)。然后将这些肽单独或混合(MIX)在完全和不完全弗氏佐剂中乳化,并皮下注射给金黄地鼠。用 P1 处理的仓鼠血清中产生的抗体可识别 蛋白的 76-150kDa 之间的蛋白。用 P2 和 P3 肽以及 MIX 处理的仓鼠血清中产生的抗体可识别 蛋白的 52-76kDa 之间的蛋白。对蛋白数据库中肽序列相似性的研究表明,它们与一种 63kDa 糖蛋白相匹配。免疫测定法(ELISA)显示,三种肽及其 MIX 被 CL 患者的血清识别。肽的 MIX 表现出最佳的性能(79%的敏感性),其次是 P1(72%的敏感性),AS 则表现出 91%的敏感性。这些结果为发现用于诊断或用于利什曼病免疫保护的分子提供了新途径。
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