Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Laboratorio de Química Orgânica Sintética, Instituto de Química, Universidade de Campinas, Campinas, Brazil.
Acta Trop. 2022 Aug;232:106521. doi: 10.1016/j.actatropica.2022.106521. Epub 2022 May 17.
Diagnosis of tegumentary leishmaniasis (TL) is essential to avoid permanent damage and severe functional sequelae and there is an urgent need to discover new antigens. The present study aimed to comprehensively evaluate the potential use of the Tryparedoxin Peroxidase (TryP) as an antigen for serological tests. The proposal integrates data from immunoproteomics with immunoinformatics, in addition to a precise analysis of protein levels in the evolutionary stages of the parasite by flow cytometry. To evaluate the performance in the diagnosis of TL, Enzyme-Linked Immunosorbent Assay (ELISA) assays were performed using the recombinant protein and the respective B-cell epitope, followed by an analysis of the contribution of this peptide in the recognition of the protein by patients, evaluated by serum depletion assays. We showed that the TryP has a linear B-cell epitope with high divergence compared to orthologs from Trypanosoma cruzi and Homo sapiens. The results also show high expression and positive cells for TryP (TryP) in the infective metacyclic promastigotes (MET) and intracellular (24 and 48 hours) stages. From the depletion assays, it was possible to confirm the contribution of the peptide in the specific recognition of the TryP protein by patients with TL (13.7-15.9%). ELISA using the peptide showed high performance in the diagnosis compared to the recombinant TryP (rTryP), Soluble Leishmania braziliensis Antigen (sLba) and Immunofluorescence Assay (IFA) with accuracy of 94.29, 89.29, 65.00 and 37.14%, respectively). We can conclude that the MNEPAPP peptide is a potential antigen for the diagnosis of TL.
皮肤利什曼病（TL）的诊断对于避免永久性损伤和严重的功能后遗症至关重要，因此迫切需要发现新的抗原。本研究旨在全面评估 Tryparedoxin Peroxidase（TryP）作为血清学检测抗原的潜在用途。该提案整合了免疫蛋白质组学和免疫信息学的数据，以及通过流式细胞术对寄生虫进化阶段的蛋白质水平进行精确分析。为了评估该方法在 TL 诊断中的性能，我们使用重组蛋白和相应的 B 细胞表位进行了酶联免疫吸附试验（ELISA）检测，随后通过血清耗竭试验分析了该肽段在患者识别蛋白中的作用。我们发现，与 Trypanosoma cruzi 和 Homo sapiens 的同源物相比，TryP 具有线性 B 细胞表位，高度分化。结果还显示，在感染性的循环前体鞭毛体（MET）和细胞内（24 和 48 小时）阶段，TryP（TryP）的表达水平和阳性细胞较高。从耗竭试验中可以确认该肽段在患者对 TL 的特异性识别 TryP 蛋白中的作用（13.7-15.9%）。与重组 TryP（rTryP）、可溶性巴西利什曼原虫抗原（sLba）和免疫荧光分析（IFA）相比，使用该肽段进行的 ELISA 检测在诊断中的性能更高，准确性分别为 94.29%、89.29%、65.00%和 37.14%）。我们可以得出结论，MNEPAPP 肽是诊断 TL 的潜在抗原。
