Center of Excellence for Antibody Research (CEAR), Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Center of Excellence for Antibody Research (CEAR), Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Biochem Biophys Res Commun. 2014 Apr 4;446(2):475-80. doi: 10.1016/j.bbrc.2014.02.131. Epub 2014 Mar 14.
Dengue virus (DENV), a re-emerging virus, constitutes the largest vector-borne disease virus, with 50-100 million cases reported every year. Although DENV infection induces lifelong immunity against viruses of the same serotypes, the subsequent infection with the heterologous serotypes can cause more severe form of the disease, such as Dengue Haemorrhagic Fever (DHF) or Dengue Shock Syndrome (DSS). However, there is neither approved vaccine nor specific drugs available to treat this disease. In this study, previously developed 19 human monoclonal antibodies (HuMAbs) showing strong to moderate cross neutralizing activity were selected. Most of them (13/19) were targeted to domain II of envelop glycoprotein. To understand and clarify the recognition properties, the maturation mechanisms comprising Variable/Diversity/Joining (VDJ) recombination, Variable Heavy (VH)/Variable Light (VL) chain pairing, variability at junctional site, and somatic hypermutation (SHM) of those antibodies were studied and compared with their predecessor germline sequences. IMGT/V-QUEST database was applied to analyze the isolated VH and VL sequences. To confirm the correction of isolated VH/VL, 3 HuMAbs (1A10H7, 1B3B9, 1G7C2) was transiently expressed in HEK293T cell. All three clones of the expressed recombinant IgG (rIgG) showed the same binding and neutralizing activity as same as those from hybridomas. The data obtained in this study will elucidate the properties of those HuMAbs for further genetic modification, and its binding epitopes.
登革病毒（DENV）是一种重新出现的病毒，是最大的虫媒病毒，每年报告的病例有 5000 万至 1 亿例。尽管 DENV 感染可诱导针对同型血清型病毒的终生免疫力，但随后感染异型血清型可导致更严重的疾病形式，如登革出血热（DHF）或登革休克综合征（DSS）。然而，目前既没有批准的疫苗，也没有专门用于治疗这种疾病的药物。在这项研究中，选择了以前开发的 19 种具有强至中度交叉中和活性的人源单克隆抗体（HuMAbs）。其中大多数（13/19）针对包膜糖蛋白的结构域 II。为了了解和阐明识别特性，研究并比较了这些抗体的成熟机制，包括可变/多样性/连接（VDJ）重组、可变重（VH）/可变轻（VL）链配对、连接点的变异性和体细胞超突变（SHM）。应用 IMGT/V-QUEST 数据库分析分离的 VH 和 VL 序列。为了确认分离的 VH/VL 的正确性，在 HEK293T 细胞中转瞬表达了 3 种 HuMAbs（1A10H7、1B3B9、1G7C2）。表达的重组 IgG（rIgG）的所有三个克隆都表现出与杂交瘤相同的结合和中和活性。本研究获得的数据将阐明这些 HuMAbs 的特性，用于进一步的遗传修饰及其结合表位。
