Viral Vaccines Laboratory, National Institute of Quality Control in Health, FIOCRUZ, Rio de Janeiro, Brazil.
Post-Graduation Program in Sanitary Surveillance, National Institute of Quality Control in Health, FIOCRUZ, Rio de Janeiro, Brazil.
PLoS One. 2022 May 6;17(5):e0267653. doi: 10.1371/journal.pone.0267653. eCollection 2022.
Dengue is caused by an arbovirus that belongs to the Flaviviridae family and there are four distinct, but close related, circulating serotypes. Dengue disease is of great importance for global public health, with vaccination being its main prophylactic measure. However, there is a paucity of biological models for evaluating tetravalent dengue vaccines. The aim of this study was to evaluate the susceptibility of human cell lines HEK293T and THP-1 to a commercial dengue vaccine and test the feasibility of this approach in the development of a potency assay with human cell lines, as a methodological alternative to the golden standard potency assay with VERO cells. In this context, we used a batch of the commercial vaccine Dengvaxia® (CYD-TDV) for the infection tests. We evaluated the presence of the vaccine virus in THP-1 cells, differentiated into macrophages (dTHP-1), and in HEK293T by confocal microscopy, using 4G2 pan-flavivirus antibody. Vaccine infectivity and potency were determined by immunocolorimetric assay using monoclonal antibodies specific for each serotype. The results indicated that the human strain HEK293T was responsive to the tetravalent vaccine, as shown by the presence of virus particles in the cell cytoplasm in a pattern similar to the one observed with VERO cells. Moreover, it was possible to determine the infectivity and potency values of each vaccine virus serotype in the HEK293T, with serotype 4 prevailing over the others. Thus, the human cell line HEK293T provides a potential candidate to be used in assays to determine potency and identity of tetravalent dengue vaccines.
登革热是由黄病毒科(Flaviviridae)的一种虫媒病毒引起的,目前有四种不同但密切相关的循环血清型。登革热疾病对全球公共卫生非常重要,疫苗接种是其主要的预防措施。然而,目前缺乏用于评估四价登革热疫苗的生物模型。本研究旨在评估人源细胞系 HEK293T 和 THP-1 对商业登革热疫苗的敏感性,并测试用人源细胞系开发效力测定的这种方法的可行性,作为替代金标准用 VERO 细胞进行效力测定的方法。在这种情况下,我们使用了一批商业疫苗 Dengvaxia®(CYD-TDV)进行感染测试。我们使用针对每种血清型的特异性单克隆抗体,通过共聚焦显微镜使用 4G2 泛黄病毒抗体,评估疫苗病毒在分化为巨噬细胞的 THP-1 细胞(dTHP-1)和 HEK293T 中的存在情况。通过免疫比色法测定使用针对每种血清型的特异性单克隆抗体,评估疫苗的感染力和效力。结果表明,人源细胞系 HEK293T 对四价疫苗有反应,其细胞质中存在病毒颗粒,与 VERO 细胞观察到的模式相似。此外,还可以确定 HEK293T 中每种疫苗病毒血清型的感染力和效力值,其中血清型 4 占优势。因此,人源细胞系 HEK293T 为用于测定四价登革热疫苗效力和鉴定的试验提供了一个潜在的候选者。