Machado Luiz Fernando Almeida, Filho Luiz Ricardo Goulart, Santos Fabiana Almeida Araújo, Siravenha Leonardo Quintão, Silva Andrea Nazaré Monteiro Rangel, Queiroz Maria Alice Freitas, Vallinoto Antonio Carlos Rosário, Ishak Marluísa Oliveira Guimarães, Ishak Ricardo
Biology of Infectious and Parasitic Agents Post-Graduate Program, Federal University of Pará, Belém, Brazil.
Virology Laboratory, Institute of Biological Sciences, Federal University of Pará, Belem, Brazil.
Front Med (Lausanne). 2022 Jun 9;9:884738. doi: 10.3389/fmed.2022.884738. eCollection 2022.
Human T-lymphotropic virus 1 (HTLV-1) is endemic worldwide and the infection results in severe diseases, including Adult T-cell Leukemia (ATL) and HTLV-1 associated myelopathy (HAM). There are some limitations of employing the present commercial serological assays for both diagnostic and epidemiological purposes in different geographical areas of the Brazil, such as the Amazon Region. Currently, methods for diagnosis are usually expensive to adapt for routine use. The aim of this work was to identify and characterize specific ligands to IgG that mimic HTLV-1 epitopes through the Phage Display technique, which could be used for diagnosis and as future vaccine candidates. Initially, IgG from 10 patients with HTLV-1 and 20 negative controls were covalently coupled to protein G-magnetic beads. After biopanning, genetic sequencing, bioinformatics analysis and Phage-ELISA were performed. The technique allowed the identification of 4 clones with HTLV-1 mimetic peptides, three aligned with gp46, A6 (SPYW), B6 (SQLP) and D7 (PLIL), and one with the protease and Tax, A8 (SPPR). Clones A6 and B6 showed higher values of accessibility, antigenicity and hydrophilicity. The reactivity of the clones evaluated by the Receiver Operating Characteristic (ROC) curve showed that the B6 clone had the highest Area Under Curve (0.83) and sensitivity and specificity values (both were 77.27 %; < 0.001). The study showed that the Phage Display technique is effective for the identification of HTLV-1-related peptides. Clone B6 indicated to be a good marker for bioprospecting diagnostic test for HTLV-1 infection and could be used as a possible vaccine candidate for future studies.
人类嗜T淋巴细胞病毒1型（HTLV-1）在全球范围内流行，该感染会导致严重疾病，包括成人T细胞白血病（ATL）和HTLV-1相关脊髓病（HAM）。在巴西的不同地理区域，如亚马逊地区，使用现有的商业血清学检测方法进行诊断和流行病学研究存在一些局限性。目前，诊断方法通常成本高昂，难以适用于常规使用。这项工作的目的是通过噬菌体展示技术鉴定和表征模拟HTLV-1表位的IgG特异性配体，这些配体可用于诊断并作为未来的疫苗候选物。最初，将来自10名HTLV-1患者和20名阴性对照的IgG共价偶联到蛋白G磁珠上。经过生物淘选后，进行了基因测序、生物信息学分析和噬菌体酶联免疫吸附测定。该技术鉴定出4个带有HTLV-1模拟肽的克隆，其中三个与gp46序列一致，分别是A6（SPYW）、B6（SQLP）和D7（PLIL），另一个与蛋白酶和Tax序列一致，即A8（SPPR）。克隆A6和B6显示出较高的可及性、抗原性和亲水性值。通过受试者工作特征（ROC）曲线评估克隆的反应性表明，B6克隆的曲线下面积最高（0.83），敏感性和特异性值也最高（均为77.27%；<0.001）。研究表明，噬菌体展示技术对于鉴定HTLV-1相关肽是有效的。克隆B6表明是用于HTLV-1感染生物勘探诊断测试的良好标志物，并且可作为未来研究的潜在疫苗候选物。
