Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses

Schmidt, Fabian; Weisblum, Yiska; Muecksch, Frauke; Hoffmann, Hans-Heinrich; Michailidis, Eleftherios; Lorenzi, Julio C. C.; Mendoza, Pilar; Rutkowska, Magdalena; Bednarski, Eva; Gaebler, Christian; Agudelo, Marianna; Cho, Alice; Wang, Zijun; Gazumyan, Anna; Cipolla, Melissa; Caskey, Marina; Robbiani, Davide F.; Nussenzweig, Michel C.; Rice, Charles M.; Hatziioannou, Théodora; Bieniasz, Paul D.

doi:10.1084/jem.20201181

Cited by 528 publications

(483 citation statements)

References 47 publications

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“…2a and Extended Data Fig. 2a ), when entry was probed using vesicular stomatitis virus (VSV) pseudoparticles carrying the SARS-CoV-2 spike protein and expressing the green fluorescent protein (GFP) 14 . However, when ACE2 and TMPRSS2 were expressed together, most of the cell lines became highly susceptible to infection, with 293T, Calu-6, HuH-6, Huh-7.5, and Caco-2 cells demonstrating considerably higher infection efficiency than Vero E6 cells.…”

Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 desensitizes host cells to interferon through inhibition of the JAK-STAT pathway

Chen

Khan

et al. 2020

Preprint

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SARS-CoV-2 can infect multiple organs, including lung, intestine, kidney, heart, liver, and brain. The molecular details of how the virus navigates through diverse cellular environments and establishes replication are poorly defined. Here, we performed global proteomic analysis of the virus-host interface in a newly established panel of phenotypically diverse, SARS-CoV-2-infectable human cell lines representing different body organs. This revealed universal inhibition of interferon signaling across cell types following SARS-CoV-2 infection. We performed systematic analyses of the JAK-STAT pathway in a broad range of cellular systems, including immortalized cell lines and primary-like cardiomyocytes, and found that several pathway components were targeted by SARS-CoV-2 leading to cellular desensitization to interferon. These findings indicate that the suppression of interferon signaling is a mechanism widely used by SARS-CoV-2 in diverse tissues to evade antiviral innate immunity, and that targeting the viral mediators of immune evasion may help block virus replication in patients with COVID-19.

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Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 desensitizes host cells to interferon through inhibition of the JAK-STAT pathway

Chen

Khan

et al. 2020

Preprint

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“…2). Previous studies demonstrated that the neutralizing activity of convalescent plasma measured with this method correlates quantitatively with neutralizing activity measured using an authentic SARS-CoV-2 neutralization assay (18,19). Neutralizing activity against SARS-CoV-2 WT or D614G S glycoprotein, as measured by the neutralization half-maximum inhibitory dilution (ID 50 ), was detected in 71% of patients 6 weeks after the onset of symptoms.…”

mentioning

confidence: 85%

Decline of Humoral Responses against SARS-CoV-2 Spike in Convalescent Individuals

Beaudoin-Bussières

Laumaea

Anand

et al. 2020

mBio

184

111

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In the absence of effective vaccines and with limited therapeutic options, convalescent plasma is being collected across the globe for potential transfusion to coronavirus disease 2019 (COVID-19) patients. The therapy has been deemed safe, and several clinical trials assessing its efficacy are ongoing. While it remains to be formally proven, the presence of neutralizing antibodies is thought to play a positive role in the efficacy of this treatment. Indeed, neutralizing titers of ≥1:160 have been recommended in some convalescent plasma trials for inclusion. Here, we performed repeated analyses at 1-month intervals on 31 convalescent individuals to evaluate how the humoral responses against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Spike glycoprotein, including neutralization, evolve over time. We observed that the levels of receptor-binding-domain (RBD)-specific IgG and IgA slightly decreased between 6 and 10 weeks after the onset of symptoms but that RBD-specific IgM levels decreased much more abruptly. Similarly, we observed a significant decrease in the capacity of convalescent plasma to neutralize pseudoparticles bearing wild-type SARS-CoV-2 S or its D614G variant. If neutralization activity proves to be an important factor in the clinical efficacy of convalescent plasma transfer, our results suggest that plasma from convalescent donors should be recovered rapidly after resolution of symptoms. IMPORTANCE While waiting for an efficient vaccine to protect against SARS-CoV-2 infection, alternative approaches to treat or prevent acute COVID-19 are urgently needed. Transfusion of convalescent plasma to treat COVID-19 patients is currently being explored; neutralizing activity in convalescent plasma is thought to play a central role in the efficacy of this treatment. Here, we observed that plasma neutralization activity decreased a few weeks after the onset of the symptoms. If neutralizing activity is required for the efficacy of convalescent plasma transfer, our results suggest that convalescent plasma should be recovered rapidly after the donor recovers from active infection.

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“…Infectivity of virions was determined by titration on 293T ACE2 cells. Further details are described elsewhere 33 .…”

Section: Sars-cov-2 and Sars-cov Pseudotyped Reporter Virusesmentioning

confidence: 99%

Convergent antibody responses to SARS-CoV-2 in convalescent individuals

Robbiani

Gaebler

Muecksch

et al. 2020

Nature

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1,894

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2,651

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During the coronavirus disease-2019 (COVID-19) pandemic, severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2) has led to the infection of millions of people and has claimed hundreds of thousands of lives. The entry of the virus into cells depends on the receptor-binding domain (RBD) of the spike (S) protein of SARS-CoV-2. Although there is currently no vaccine, it is likely that antibodies will be essential for protection. However, little is known about the human antibody response to SARS-CoV-2 1-5. Here we report on 149 COVID-19-convalescent individuals. Plasma samples collected an average of 39 days after the onset of symptoms had variable half-maximal pseudovirus neutralizing titres; titres were less than 50 in 33% of samples, below 1,000 in 79% of samples and only 1% of samples had titres above 5,000. Antibody sequencing revealed the expansion of clones of RBD-specific memory B cells that expressed closely related antibodies in different individuals. Despite low plasma titres, antibodies to three distinct epitopes on the RBD neutralized the virus with half-maximal inhibitory concentrations (IC 50 values) as low as 2 ng ml −1. In conclusion, most convalescent plasma samples obtained from individuals who recover from COVID-19 do not contain high levels of neutralizing activity. Nevertheless, rare but recurring RBD-specific antibodies with potent antiviral activity were found in all individuals tested, suggesting that a vaccine designed to elicit such antibodies could be broadly effective.

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Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses

Cited by 528 publications

References 47 publications

SARS-CoV-2 desensitizes host cells to interferon through inhibition of the JAK-STAT pathway

SARS-CoV-2 desensitizes host cells to interferon through inhibition of the JAK-STAT pathway

Decline of Humoral Responses against SARS-CoV-2 Spike in Convalescent Individuals

Convergent antibody responses to SARS-CoV-2 in convalescent individuals

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