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新型基因构建体用于生产重组 HTLV-1/2 抗原,并评估其对 HTLV-1 感染患者血浆样本的反应性。

Novel Genetic Constructs for Production of Recombinant HTLV-1/2 Antigens and Evaluation of Their Reactivity to Plasma Samples from HTLV-1-Infected Patients.

机构信息

Laboratory of Structural Biology and Protein Engineering, Carlos Chagas Institute, Oswaldo Cruz Foundation, Curitiba, Paraná, Brazil.

Cellular and Molecular Biology Postgraduate Program, Federal University of Paraná, Curitiba, Paraná, Brazil.

出版信息

J Clin Microbiol. 2021 Mar 19;59(4). doi: 10.1128/JCM.02701-20.

Abstract

Human T-cell leukemia virus type 1 (HTLV-1) can cause life-threatening diseases for which there are no effective treatments. Prevention of HTLV-1 infection requires massive testing of pregnant women, blood for transfusion, and organs for transplantation, as well as safe sex. In this context, serological assays are widely used for monitoring HTLV-1 infections. Despite the necessity for recombinant antigens to compose serological tests, there is little information available on procedures to produce recombinant HTLV-1/2 antigens for serological diagnostic purposes. In this work, we tested a series of genetic constructions to select those more amenable for production in bacterial systems. To overcome the constraints in expressing sections of viral envelope proteins in bacteria, we have used the p24 segment of the gag protein as a scaffold to display the immunogenic regions of gp46 and gp21. Nine recombinant antigenic proteins derived from HTLV-1 and five derived from HTLV-2 were successfully purified. The HTLV-1 antigens showed high efficiency in discriminating HTLV-positive samples from HTLV-negative samples using enzyme-linked immunosorbent assays. Interestingly, HTLV-1-positive samples showed a high level of cross-reaction with HTLV-2 antigens. This finding is explained by the high sequence conservation between the structural proteins of these two highly related viruses. In summary, the results presented in this work provide a detailed description of the methods used to produce recombinant HTLV-1 and HTLV-2 antigens, and they demonstrate that the HTLV-1 antigens show strong potential for serological diagnosis of HTLV-1 infections.

摘要

人类 T 细胞白血病病毒 1 型(HTLV-1)可引起危及生命的疾病,目前尚无有效治疗方法。预防 HTLV-1 感染需要对孕妇、输血和移植用器官进行大规模检测,同时还需要安全性行为。在这种情况下,血清学检测被广泛用于监测 HTLV-1 感染。尽管需要使用重组抗原来构成血清学检测,但关于生产用于血清学诊断目的的重组 HTLV-1/2 抗原的程序信息很少。在这项工作中,我们测试了一系列遗传构建体,以选择更适合在细菌系统中生产的构建体。为了克服在细菌中表达病毒包膜蛋白部分的限制,我们使用 gag 蛋白的 p24 片段作为支架来展示 gp46 和 gp21 的免疫原性区域。成功纯化了 9 种源自 HTLV-1 的重组抗原蛋白和 5 种源自 HTLV-2 的重组抗原蛋白。使用酶联免疫吸附试验,这些 HTLV-1 抗原能够高效地区分 HTLV-阳性样本和 HTLV-阴性样本。有趣的是,HTLV-1 阳性样本与 HTLV-2 抗原表现出高度的交叉反应。这一发现可以用这两种高度相关的病毒的结构蛋白之间的高序列保守性来解释。总之,本工作中提出的结果提供了生产重组 HTLV-1 和 HTLV-2 抗原的详细描述,并且表明 HTLV-1 抗原具有用于 HTLV-1 感染的血清学诊断的强大潜力。

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