A pseudovirus system for the testing of antiviral activity of compounds in different cell lines

Чересиз, С. В.; Grigoryev, I. V.; Семенова, Е. А.; Pustylnyak, Vladimir O.; Власов, В. В.; Pokrovsky, Andrey G.

doi:10.1134/s1607672910060049

Cited by 6 publications

(3 citation statements)

References 6 publications

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“…Furthermore, they are manual, labor-intensive assays with turn-around-times of several days. Finally, for lentivirus based PNAs, serum and plasma from patients receiving antiretroviral therapy or pre-exposure prophylaxis for HIV may contain inhibitors of pseudovirus activity non-specific to SARS-CoV-2 22 .…”

Section: Introductionmentioning

confidence: 99%

Rapid, High Throughput, Automated Detection Of SARS-Cov-2 Neutralizing Antibodies Against Native-Like Vaccine And Delta Variant Spike Trimers

Cheedarla

Verkerke

Potlapalli

et al. 2022

Preprint

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Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of hACE-2 binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using vaccine and delta variant viral strains showed strong correlation with cell-based pseudovirus and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta variant resistance to neutralization in samples with paired vaccine and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.

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Section: Introductionmentioning

confidence: 99%

Rapid, High Throughput, Automated Detection Of SARS-Cov-2 Neutralizing Antibodies Against Native-Like Vaccine And Delta Variant Spike Trimers

Cheedarla

Verkerke

Potlapalli

et al. 2022

Preprint

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show abstract

“…In recent years, many researchers give preference to the pseudotyped virus approach, a safer method suitable for BSL-2 lab settings (Li Q. et al, 2018;Montefiori et al, 2018). Compared to viral isolates and infectious molecular clones, pseudotyped viruses are harmless, because virus replication is restricted to a single cycle due to mutations in coding regions of the genome, which is why pseudotyped viruses are often called single-cycle viruses (Cheresiz et al, 2010;Li Q. et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Model systems of human immunodef iciency virus (HIV-1) for in vitro eff icacy assessment of candidate vaccines and drugs against HIV-1

et al. 2022

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HIV infection still remains a major challenge for healthcare systems of the world. There are several aspects on counteracting the HIV/AIDS epidemic. The f irst aspect covers preventive measures including educational campaigns on HIV/AIDS and promotion of a healthy lifestyle, protected sex, and pre-exposure prophylaxis of vulnerable groups. The second aspect is timely HIV testing and the use of antiretroviral therapy when test results come back positive. The third aspect is the scientif ic research associated with discovering new pharmaceutical agents and developing HIV-1 vaccines. Selecting an adequate tool for quick and accurate in vitro eff icacy assessment is the key aspect for eff icacy assessment of vaccines and chemotherapy drugs. The classical method of virology, which makes it possible to evaluate the neutralizing activity of the sera of animals immunized with experimental vaccines and the eff icacy of chemotherapy agents is the method of neutralization using viral isolates and infectious molecular clones, i. e. infectious viral particles obtained via cell transfection with a plasmid vector including the full-length HIV-1 genome coding structural, regulatory, and accessory proteins of the virus required for the cultivation of replication-competent viral particles in cell culture. However, neutralization assessment using viral isolates and infectious molecular clones is demanding in terms of time, effort, and biosafety measures. An alternative eliminating these disadvantages and allowing for rapid screening is the use of pseudoviruses, which are recombinant viral particles, for the analysis of neutralizing activity. Pseudotyped viruses have defective genomes restricting their replication to a single cycle, which renders them harmless compared to infectious viruses. The present review focuses on describing viral model systems for in vitro eff icacy assessment of vaccines and drugs against HIV-1, which include primary HIV-1 isolates, laboratoryadapted strains, infectious molecular clones, and env-pseudoviruses. A brief comparison of the listed models is presented. The HIV-1 env-pseudoviruses approach is described in more detail.

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“…Lentiviral vectors have been used in our laboratory, as well as in other laboratories, in order to design safe systems for the screening of inhibitors of wild-type HIV-1 replication [14–18]. These systems are represented by a recombinant lentivirus carrying a fragment of the HIV-1 genome, without the regions that encode virus peptides and contain the gene of a reporter (marker) protein (e.g., green fluorescent protein).…”

Section: Introductionmentioning

confidence: 99%

Screening of Potential HIV-1 Inhibitors/ Replication Blockers Using Secure Lentiviral in Vitro System

et al. 2011

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The development and usage of safe cell systems for testing agents which possess anti-HIV activity is a very important factor in the design of new drugs. We have described in detail a system we designed that is based on lentiviral vectors (Prokofjeva et. al.,Antiviral Therapy,in print) for swift and completely safe screening of potential HIV-1 replication inhibitors. The system enables one to test the efficiency of the inhibitory activity of compounds whose action is directed towards either wild-type HIV-1 reverse transcriptase or integrase, or mutant enzymes corresponding to the drug-resistant virus form. Testing results of a number of already known drugs, which correlate well with published data as well as data on newly synthesized compounds, were obtained. Application of this system substantially broadens the possibilities of preclinical anti-HIV drugs testing.

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A pseudovirus system for the testing of antiviral activity of compounds in different cell lines

Cited by 6 publications

References 6 publications

Rapid, High Throughput, Automated Detection Of SARS-Cov-2 Neutralizing Antibodies Against Native-Like Vaccine And Delta Variant Spike Trimers

Rapid, High Throughput, Automated Detection Of SARS-Cov-2 Neutralizing Antibodies Against Native-Like Vaccine And Delta Variant Spike Trimers

Model systems of human immunodef iciency virus (HIV-1) for in vitro eff icacy assessment of candidate vaccines and drugs against HIV-1

Screening of Potential HIV-1 Inhibitors/ Replication Blockers Using Secure Lentiviral in Vitro System

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