Monitoring SARS-CoV-2 Infection Using a Double Reporter-Expressing Virus

Chiem, Kevin; Park, Jun-Gyu; Vasquez, Desarey Morales; Plemper, Richard K.; Torrelles, Jordi B.; Kobie, James J.; Walter, Mark R.; Ye, Chengjin; Martínez-Sobrido, Luis

doi:10.1128/spectrum.02379-22

Cited by 9 publications

(6 citation statements)

References 75 publications

(147 reference statements)

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“…To further test whether our vaccine regiment could induce protective immunity against live virus infection, we performed a challenge study using recombinant SARS-CoV-2 expressing mCherry and nanoluciferase (rSARS-CoV-2/mCherry-Nluc) at a dose of 10 4 PFU/mouse (39). fK18 hACE2 transgenic mice were first primed with either PBS or inactivated PR8 virus and then subsequently vaccinated with either NP/RBD or RBD and challenged with rSARS-CoV-2 (Fig.…”

Section: Np/rbd Vaccination In Mice Preexposed To Inactivated Pr8 Pro...mentioning

confidence: 99%

“…SARS-CoV-2 neutraliza tion assays were performed with USA-WA1/2020 SARS-CoV-2 strain. The rSARS-CoV-2/ mCherry-Nluc was previously described (39). Viral stocks were generated in Vero E6 cells, aliquoted, titrated, sequenced and stored at −80°C as previously described (39).…”

Section: Virus Strainsmentioning

confidence: 99%

“…For viral challenge studies, groups of vaccinated K18 hACE2 transgenic mice were anesthetized with gaseous isoflurane and inoculated intranasally with 10 4 PFU/mouse of rSARS-CoV-2/mCherry-Nluc (39). A separate group of K18 hACE2 transgenic mice were also mock-infected with PBS and served as a negative control.…”

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

“…Mice that were below 75% of their initial body weight were considered to have reached their experimental endpoint and were humanly euthanized. In vivo bioluminescence imaging of live mice (n = 3) was conducted with an Ami HT in vivo imaging system (IVIS; Spectral Instruments) at 2 and 4 DPI (39). Mice were anesthe tized with isoflurane and retro-orbitally injected with 100 µL of Nano-Glo luciferase substrate diluted by 1:10 in PBS (39).…”

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

“…In vivo bioluminescence imaging of live mice (n = 3) was conducted with an Ami HT in vivo imaging system (IVIS; Spectral Instruments) at 2 and 4 DPI (39). Mice were anesthe tized with isoflurane and retro-orbitally injected with 100 µL of Nano-Glo luciferase substrate diluted by 1:10 in PBS (39). Mice were immediately placed in an isolation chamber and imaged using the Ami HT IVIS.…”

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

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Harnessing preexisting influenza virus-specific immunity increases antibody responses against SARS-CoV-2

Dulin,

Barre,

et al. 2024

J Virol

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In pandemic scenarios involving novel human pathogenic viruses, it is highly desirable that vaccines induce strong neutralizing antibodies as quickly as possible. However, current vaccine strategies require multiple immunization doses to produce high titers of neutralizing antibodies and are poorly protective after a single vaccination. We therefore wished to design a vaccine candidate that would induce increased protective immune responses following the first vaccine dose. We hypothesized that antibodies against the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein could be increased by drawing upon immunity to a previous infection. We generated a fusion protein containing the influenza H1N1 PR8 virus nucleoprotein (NP) and the SARS-CoV-2 spike RBD. Mice with or without preexisting immunity to PR8 were then vaccinated with NP/RBD. We observed significantly increased SARS-CoV-2 neutralizing antibodies in mice with PR8 immunity compared to mice without preexisting PR8 immunity. Vaccination with NP/RBD protected mice from SARS-CoV-2-induced morbidity and mortality after a single dose. Additionally, we compared SARS-CoV-2 virus titers in the lungs and nasal turbinates 4 days post-challenge of mice vaccinated with NP/RBD. SARS-CoV-2 virus was detectable in the lungs and nasal turbinate of mice without preexisting PR8 immunity, while SARS-CoV-2 virus was completely undetectable in mice with preexisting PR8 immunity. We also found that CD4-positive T cells in mice with preexisting immunity to PR8 play an essential role in producing the increased antibody response against RBD. This vaccine strategy potentially can be modified to target other pathogens of concern and offers extra value in future pandemic scenarios. IMPORTANCE Increased globalization and changes in human interactions with wild animals has increased the likelihood of the emergence of novel viruses with pandemic potential. Vaccines can be effective in preventing severe disease caused by pandemic viruses. However, it takes time to develop protective immunity via prime-boost vaccination. More effective vaccine designs should quickly induce protective immunity. We propose leveraging preexisting immunity to a different pathogen to boost protection against emerging viruses. We targeted SARS-CoV-2 as a representative pandemic virus and generated a fusion protein vaccine that combines the nucleoprotein from influenza A virus and the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Our vaccine design significantly increased the production of RBD-specific antibodies in mice that had previously been exposed to influenza virus, compared to those without previous exposure. This enhanced immunity reduced SARS-CoV-2 replication in mice. Our results offer a vaccine design that could be valuable in a future pandemic setting.

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Section: Np/rbd Vaccination In Mice Preexposed To Inactivated Pr8 Pro...mentioning

confidence: 99%

Section: Virus Strainsmentioning

confidence: 99%

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

Section: Mouse Sars-cov-2 Challengementioning

confidence: 99%

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