Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, 1417613151, Iran.
Research Centre for Emerging and Reemerging Infectious Diseases, Pasteur Institute of Iran, Tehran, Iran.
Arch Virol. 2020 May;165(5):1109-1120. doi: 10.1007/s00705-020-04576-9. Epub 2020 Mar 18.
Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne disease with a mortality rate of up to 50% in humans. To avoid safety concerns associated with the use of live virus in virus neutralization assays and to detect human serum neutralizing antibodies, we prepared lentiviral particles containing the CCHF glycoprotein (lenti-CCHFV-GP). Incorporation of the GP into the lentiviral particle was confirmed by electron microscopy and Western blotting. Lenti-CCHFV-GP was found to be able to infect a wide range of cell lines, including BHK-21, HeLa, HepG2, and AsPC-1 cells. In addition, lenti-CCHFV-GP was successfully used as an alternative to CCHFV for the detection of neutralizing antibodies. Sera collected from CCHF survivors neutralized lenti-CCHFV-GP particles in a dose-dependent manner. Our results suggest that the lenti-CCHFV-GP pseudovirus can be used as a safe tool for neutralization assays in low-containment laboratories.
克里米亚-刚果出血热（CCHF）是一种蜱媒疾病，人类死亡率高达 50%。为了避免与在病毒中和测定中使用活病毒相关的安全问题，并检测人血清中和抗体，我们制备了含有 CCHF 糖蛋白的慢病毒颗粒（lenti-CCHFV-GP）。通过电子显微镜和 Western blot 证实了 GP 整合到慢病毒颗粒中。发现 lenti-CCHFV-GP 能够感染多种细胞系，包括 BHK-21、Hela、HepG2 和 AsPC-1 细胞。此外，lenti-CCHFV-GP 可成功替代 CCHFV 用于检测中和抗体。从 CCHF 幸存者中采集的血清以剂量依赖的方式中和 lenti-CCHFV-GP 颗粒。我们的结果表明，lenti-CCHFV-GP 假病毒可作为低生物安全实验室中中和测定的安全工具。
