Department of Microbiology, National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA 02115, USA.
Department of Immunology, Harvard Medical School, Boston, MA 02115, USA.
Viruses. 2022 Dec 23;15(1):43. doi: 10.3390/v15010043.
Viral pathogens with the potential to cause widespread disruption to human health and society continue to emerge or re-emerge around the world. Research on such viruses often involves high biocontainment laboratories (BSL3 or BSL4), but the development of diagnostics, vaccines and therapeutics often uses assays that are best performed at lower biocontainment. Reliable inactivation is necessary to allow removal of materials to these spaces and to ensure personnel safety. Here, we validate the use of gamma irradiation to inactivate culture supernatants and pellets of cells infected with a representative member of the Filovirus and Coronavirus families. We show that supernatants and cell pellets containing SARS-CoV-2 are readily inactivated with 1.9 MRad, while Ebola virus requires higher doses of 2.6 MRad for supernatants and 3.8 MRad for pellets. While these doses of radiation inactivate viruses, proinflammatory cytokines that are common markers of virus infection are still detected with low losses. The doses required for virus inactivation of supernatants are in line with previously reported values, but the inactivation of cell pellets has not been previously reported and enables new approaches for analysis of protein-based host responses to infection.
具有潜在能力在全球范围内对人类健康和社会造成广泛破坏的病毒病原体不断出现或再次出现。此类病毒的研究通常涉及高生物安全实验室（BSL3 或 BSL4），但诊断、疫苗和治疗药物的开发通常使用在较低生物安全级别下效果最佳的检测方法。为了允许将材料转移到这些空间，并确保人员安全，可靠的失活是必要的。在这里，我们验证了使用伽马辐照来失活感染有代表性的丝状病毒科和冠状病毒科成员的细胞培养上清液和细胞沉淀。我们表明，含有 SARS-CoV-2 的上清液和细胞沉淀很容易被 1.9 MRad 失活，而埃博拉病毒需要更高的剂量，上清液为 2.6 MRad，沉淀为 3.8 MRad。虽然这些辐射剂量可以使病毒失活，但仍然可以检测到低水平的作为病毒感染常见标志物的促炎细胞因子。上清液中病毒失活所需的剂量与之前报道的值一致，但细胞沉淀的失活尚未得到报道，这为分析基于蛋白质的宿主对感染的反应提供了新的方法。
