Virus Inactivation by Formaldehyde and Common Lysis Buffers

Seeburg, Ulrike; Urda, Lorena; Otte, Fabian; Lett, Martin J.; Caimi, S.; Mittelholzer, Christian; Klimkait, Thomas

doi:10.3390/v15081693

Cited by 3 publications

References 10 publications

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Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Ladinsky,

Zhu,

Ullah

et al. 2024

Preprint

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HIV-1 delivers its genetic material to infect a cell after fusion of the viral and host cell membranes, which happens after the viral envelope (Env) binds host receptor and co-receptor. Binding host receptor CD4 to Env results in conformational changes that allow interaction with a co-receptor (CCR5 or CXCR4). Further conformational rearrangements result in an elongated pre-hairpin intermediate structure in which Env is anchored to the viral membrane by its transmembrane region and to the host cell membrane by its fusion peptide. Although budding virions can be readily imaged by ET of HIV-1–infected tissues and cultured cells, fusing virions (attached to host cells via pre-hairpin intermediates) are not visualized, perhaps because membrane fusion is too fast to capture by EM. To image fusing virions, we used fusion inhibitors to prevent conformational changes in Env that lead to membrane fusion, thereby trapping virions linked to target cells by pre-hairpin intermediates. ET of HIV-1 pseudovirions bound to CD4+/CCR5+ TZM-bl cells revealed presumptive pre-hairpin intermediates as 2-4 narrow spokes linking a virion to the cell surface. To extend these results to a more physiological setting, we used ET to image tissues and organs derived from humanized mice infected with HIV-1 and then treated with CPT31, a high-affinity D-peptide fusion inhibitor linked to cholesterol. Trapped HIV-1 virions were found in all tissues studied (small intestine, mesenteric lymph nodes, spleen, bone marrow). Spokes representing pre-hairpin intermediates linking trapped virions to cell surfaces were similar to those seen in the previous pseudovirus/cultured cell ET study.

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Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Ladinsky,

Zhu,

Ullah

et al. 2024

Preprint

View full text Add to dashboard Cite

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Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Ladinsky,

Zhu,

Ullah

et al. 2024

J Virol

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HIV-1 delivers its genetic material to infect a cell after fusion of the viral and host cell membranes, which takes place after the viral envelope (Env) binds host receptor and co-receptor proteins. Binding of host receptor CD4 to Env results in conformational changes that allow interaction with a host co-receptor (CCR5 or CXCR4). Further conformational rearrangements result in an elongated pre-hairpin intermediate structure in which Env is anchored to the viral membrane by its transmembrane region and to the host cell membrane by its fusion peptide. Although budding virions can be readily imaged by electron tomography (ET) of HIV-1-infected tissues and cultured cells, virions that are fusing (attached to host cells via pre-hairpin intermediates) are not normally visualized, perhaps because the process of membrane fusion is too fast to capture by ET. To image virions during fusion, we used fusion inhibitors to prevent downstream conformational changes in Env that lead to membrane fusion, thereby trapping HIV-1 virions linked to target cells by pre-hairpin intermediates. ET of HIV-1 pseudovirions bound to CD4 + /CCR5 + TZM-bl cells revealed presumptive pre-hairpin intermediates as 2–4 narrow spokes linking a virion to the cell surface. To extend these results to a more physiological setting, we used ET to image tissues and organs derived from humanized bone marrow/liver/thymus mice infected with HIV-1 and then treated with CPT31, a high-affinity D-peptide fusion inhibitor linked to cholesterol. Trapped HIV-1 virions were found in all tissues studied (small intestine, mesenteric lymph nodes, spleen, and bone marrow), and spokes representing pre-hairpin intermediates linking trapped virions to cell surfaces were similar in structure and number to those seen in the previous pseudovirus and cultured cell ET study. IMPORTANCE Trapped and untrapped HIV-1 virions, both mature and immature, were distinguished by localizing spokes via 3D tomographic reconstructions of HIV-1 infected and fusion-inhibitor-treated tissues of humanized mice. The findings of trapped HIV-1 virions in all tissues examined demonstrate a wide distribution of the CPT31 inhibitor, a desirable property for a potential therapeutic. In addition, the presence of virions trapped by spokes, particularly in vascular endothelial cells, demonstrates that the fusion inhibitors can be used as markers for potential HIV-1-target cells within tissues, facilitating the mapping of HIV-1 target cells within the complex cellular milieu of infected tissues.

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Preclinical Efficacy of Cap-Dependent and Independent mRNA Vaccines against Bovine Viral Diarrhea Virus-1

Huang,

Hu,

Niu

et al. 2024

Veterinary Sciences

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Bovine viral diarrhea virus (BVDV) is an RNA virus associated with severe economic losses in animal production. Effective vaccination and viral surveillance are urgent for the prevention and control of BVDV infection. However, the application of traditional modified live vaccines and inactivated vaccines is faced with tremendous challenges. In the present study, we describe the preclinical efficacy of two BVDV mRNA vaccines tested in mice and guinea pigs, followed by a field trial in goats, where they were compared to a commercial vaccine (formaldehyde inactivated). The two mRNAs were engineered to express the envelope protein E2 of BVDV-1, the most prevalent subtype across the world, through a 5′ cap-dependent or independent fashion. Better titers of neutralizing antibodies against BVDV-1 were achieved using the capped RNA in the sera of mice and guinea pigs, with maximum values reaching 9.4 and 13.7 (by −log2), respectively, on the 35th day post-vaccination. At the same time point, the antibody levels in goats were 9.1 and 10.2 for the capped and capless RNAs, respectively, and there were no significant differences compared to the commercial vaccine. The animals remained healthy throughout the experiment, as reflected by their normal leukogram profiles. Collectively, our findings demonstrate that mRNA vaccines have good safety and immunogenicity, and we laid a strong foundation for the further exploitation of efficient and safe BVDV vaccines.

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Virus Inactivation by Formaldehyde and Common Lysis Buffers

Cited by 3 publications

References 10 publications

Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Electron tomography visualization of HIV-1 virions trapped by fusion inhibitors to host cells in infected tissues

Preclinical Efficacy of Cap-Dependent and Independent mRNA Vaccines against Bovine Viral Diarrhea Virus-1

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