Rapid Quantification of SARS-CoV-2-Neutralizing Antibodies Using Propagation-Defective Vesicular Stomatitis Virus Pseudotypes

Zettl, Ferdinand; Meister, Toni Luise; Vollmer, Tanja; Fischer, Bastian; Steinmann, Jörg; Krawczyk, Adalbert; V’kovski, Philip; Todt, Daniel; Steinmann, Eike; Pfaender, Stephanie; Zimmer, Gert

doi:10.3390/vaccines8030386

Cited by 86 publications

(98 citation statements)

References 29 publications

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“…Functional assays like virus neutralisation tests are essential to address specific questions related to protective immunity after vaccination or natural infection. However, these type of assays are operated based on in-house protocols and lack standardization across laboratories [16][17][18] . We performed an independent two-centre clinical evaluation of the GenScript cPass™ test, which is based on the principle of an inhibition ELISA, to assess its value for routine diagnostics and as surrogate functional assay to measure neutralising capability in SARS-CoV-2 elicited antibody responses.…”

Section: Discussionmentioning

confidence: 99%

Validation and clinical evaluation of a SARS-CoV-2 surrogate virus neutralisation test (sVNT)

Meyer

Reimerink²,

Torriani

et al. 2020

Emerging Microbes & Infections

134

133

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To understand SARS-CoV-2 immunity after natural infection or vaccination, functional assays such as virus neutralising assays are needed. So far, assays to detect SARS-CoV-2 neutralising antibodies rely on cellculture based infection assays either using wild type SARS-CoV-2 or pseudotyped viruses. Such assays are labour-intensive, require appropriate biosafety facilities and are difficult to standardize. Recently, a new surrogate virus neutralisation test (sVNT) was described that uses the principle of an ELISA to measure the neutralisation capacity of anti-SARS-CoV-2 antibodies directed against the receptor binding domain. Here, we performed an independent evaluation of the robustness, specificity and sensitivity on an extensive panel of sera from 269 PCR-confirmed COVID-19 cases and 259 unmatched samples collected before 2020 and compared it to cell-based neutralisation assays. We found a high specificity of 99.2 (95%CI: 96.9-99.9) and overall sensitivity of 80.3 (95%CI: 74.9-84.8) for the sVNT. Clinical sensitivity increased between early (<14 days post symptom onset or post diagnosis, dpos/dpd) and late sera (>14 dpos/dpd) from 75.0 (64.7-83.2) to 83.1 (76.5-88.1). Also, higher severity was associated with an increase in clinical sensitivity. Upon comparison with cell-based neutralisation assays we determined an analytical sensitivity of 74.3 (56.4-86.9) and 98.2 (89.4-99.9) for titres ≥10 to <40 and ≥40 to <160, respectively. Only samples with a titre ≥160 were always positive in the sVNT. In conclusion, the sVNT can be used as an additional assay to determine the immune status of COVID-19 infected of vaccinated individuals but its value needs to be assessed for each specific context.

show abstract

Section: Discussionmentioning

confidence: 99%

Validation and clinical evaluation of a SARS-CoV-2 surrogate virus neutralisation test (sVNT)

Meyer

Reimerink²,

Torriani

et al. 2020

Emerging Microbes & Infections

134

133

View full text Add to dashboard Cite

show abstract

“…Importantly, the analysis of functional activity of SARS‐CoV2‐reactive T cells showed that the majority of T cells were bi‐ or trifunctional producing Th1 proinflammatory cytokines TNFα, IFNγ, IL‐2, or granzyme B. Because such polyfunctional cells are commonly considered as indicators of protective immunity, 23,24 the detection of polyfunctionality within SARS‐CoV2‐reactive T cells points to their protective antiviral capacity.…”

Section: Discussionmentioning

confidence: 99%

“…Flow cytometry data were analyzed using FlowJo version 10.6.2 (BD Biosciences, San Jose, CA), R version 3.6.2, and Prism version 8 (GraphPad Software, LaJolla, CA). For the virus neutralization assay, a propagation‐incompetent VSV*ΔG(FLuc) pseudovirus system bearing the SARS‐CoV‐2 spike protein in the envelope was incubated with serial dilutions of immune sera prior to infections of Vero E6 cells, as described before 24 . At 18 hours post infection, firefly luciferase (FLuc) reporter activity was determined and the reciprocal antibody dilution causing 50% inhibition of the luciferase reporter calculated (PVN50).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Immune monitoring facilitates the clinical decision in multifocal COVID-19 of a pancreas-kidney transplant patient

Babel

Anft

Blázquez-Navarro

et al. 2020

American Journal of Transplantation

Self Cite

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The optimal management in transplant recipients with coronavirus disease 2019 (COVID‐19) remains uncertain. The main concern is the ability of immunosuppressed patients to generate sufficient immunity for antiviral protection. Here, we report on immune monitoring facilitating a successful outcome of severe severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2)‐associated pneumonia, meningoencephalitis, gastroenteritis, and acute kidney and pancreas graft failure in a pancreas‐kidney transplant recipient. Despite the very low numbers of circulating B, NK, and T cells identified in follow‐up, a strong SARS‐CoV‐2 reactive T cell response was observed. Importantly, we detected T cells reactive to Spike, Membrane, and Nucleocapsid proteins of SARS‐CoV‐2 with majority of T cells showing polyfunctional proinflammatory Th1 phenotype at all analyzed time points. Antibodies against Spike protein were also detected with increasing titers in follow‐up. Neutralization tests confirmed their antiviral protection. A correlation between cellular and humoral immunity was observed underscoring the specificity of demonstrated data. We conclude that analyzing the kinetics of nonspecific and SARS‐CoV‐2‐reactive cellular and humoral immunity can facilitate the clinical decision on immunosuppression adjustment and allow successful outcome as demonstrated in the current clinical case. Although the antiviral protection of the detected SARS‐CoV‐2‐reactive T cells requires further evaluation, our data prove an ability mounting a strong SARS‐CoV‐2‐reactive T cell response with functional capacity in immunosuppressed patients.

show abstract

“…Functional assays like virus neutralization tests are essential to address specific questions related to protective immunity after vaccination or natural infection. However, these type of assays are operated based on inhouse protocols and lack standardization across laboratories [16][17][18] . We performed an independent two-centre clinical evaluation of the GenScript cPass™ test, which is based on the principle of an inhibition ELISA, to assess its value for routine diagnostics and as surrogate functional assay to measure neutralizing capability in SARS-CoV-2 elicited antibody responses.…”

Section: Discussionmentioning

confidence: 99%

Validation and clinical evaluation of a SARS-CoV-2 Surrogate Virus Neutralization Test (sVNT)

Meyer

Reimerink²,

Torriani

et al. 2020

Preprint

View full text Add to dashboard Cite

To understand SARS-CoV-2 immunity after natural infection or vaccination, functional assays such as virus neutralizing assays are needed. So far, assays to determine SARS-CoV-2 neutralizing antibodies rely on cell-culture based infection assays either using wild type SARS-CoV-2 or pseudotyped viruses. Such assays are labour-intensive, require appropriate biosafety facilities and are difficult to standardize. Recently, a new surrogate virus neutralisation assay (sVNT) was described that uses the principle of an ELISA to measure the neutralization capacity of anti-SARS-CoV-2 antibodies directed against the receptor binding domain. Here, we performed an independent evaluation of the robustness, specificity and sensitivity on an extensive panel of sera from 269 PCR-confirmed COVID-19 cases and 259 unmatched samples collected before 2020 and compared it to cell-based neutralization assays. We found a high specificity of 99.2 (95%CI: 96.9-99.9) and overall sensitivity of 80.3 (95%CI: 74.9-84.8) for the sVNT. Clinical sensitivity increased between early (<14 days post symptom onset or post diagnosis, dpos/dpd) and late sera (>14 dpos/dpd) from 75.0 (64.7-83.2) to 83.1 (76.5-88.1). Also, higher severity was associated with an increase in clinical sensitivity. Upon comparison with cell-based neutralisation assays we determined an analytical sensitivity of 74.3 (56.4-86.9) and 98.2 (89.4-99.9) for titres ≥10 to <40 and ≥40 to <160, respectively. Only samples with a titre ≥160 were always positive in the sVNT. In conclusion, the sVNT can be used as an additional assay to determine the immune status of COVID-19 infected of vaccinated individuals but its value needs to be assessed for the specific context of use.

show abstract

Rapid Quantification of SARS-CoV-2-Neutralizing Antibodies Using Propagation-Defective Vesicular Stomatitis Virus Pseudotypes

Cited by 86 publications

References 29 publications

Validation and clinical evaluation of a SARS-CoV-2 surrogate virus neutralisation test (sVNT)

Validation and clinical evaluation of a SARS-CoV-2 surrogate virus neutralisation test (sVNT)

Immune monitoring facilitates the clinical decision in multifocal COVID-19 of a pancreas-kidney transplant patient

Validation and clinical evaluation of a SARS-CoV-2 Surrogate Virus Neutralization Test (sVNT)

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