Cargando…

SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine

Cell-based assays are a valuable tool for examination of virus–host cell interactions and drug discovery processes, allowing for a more physiological setting compared to biochemical assays. Despite the fact that cell-based SPR assays are label-free and thus provide all the associated benefits, they...

Descripción completa

Detalles Bibliográficos
Autores principales: Genova-Kalou, Petia, Dyankov, Georgi, Marinov, Radoslav, Mankov, Vihar, Belina, Evdokiya, Kisov, Hristo, Strijkova-Kenderova, Velichka, Kantardjiev, Todor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8392451/
https://www.ncbi.nlm.nih.gov/pubmed/34436052
http://dx.doi.org/10.3390/bios11080251
_version_ 1783743507023790080
author Genova-Kalou, Petia
Dyankov, Georgi
Marinov, Radoslav
Mankov, Vihar
Belina, Evdokiya
Kisov, Hristo
Strijkova-Kenderova, Velichka
Kantardjiev, Todor
author_facet Genova-Kalou, Petia
Dyankov, Georgi
Marinov, Radoslav
Mankov, Vihar
Belina, Evdokiya
Kisov, Hristo
Strijkova-Kenderova, Velichka
Kantardjiev, Todor
author_sort Genova-Kalou, Petia
collection PubMed
description Cell-based assays are a valuable tool for examination of virus–host cell interactions and drug discovery processes, allowing for a more physiological setting compared to biochemical assays. Despite the fact that cell-based SPR assays are label-free and thus provide all the associated benefits, they have never been used to study viral growth kinetics and to predict drug antiviral response in cells. In this study, we prove the concept that the cell-based SPR assay can be applied in the kinetic analysis of the early stages of viral infection of cells and the antiviral drug activity in the infected cells. For this purpose, cells immobilized on the SPR slides were infected with human coronavirus HCov-229E and treated with hydroxychloroquine. The SPR response was measured at different time intervals within the early stages of infection. Methyl Thiazolyl Tetrazolium (MTT) assay was used to provide the reference data. We found that the results of the SPR and MTT assays were consistent, and SPR is a reliable tool in investigating virus–host cell interaction and the mechanism of action of viral inhibitors. SPR assay was more sensitive and accurate in the first hours of infection within the first replication cycle, whereas the MTT assay was not so effective. After the second replication cycle, noise was generated by the destruction of the cell layer and by the remnants of dead cells, and masks useful SPR signals.
format Online
Article
Text
id pubmed-8392451
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-83924512021-08-28 SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine Genova-Kalou, Petia Dyankov, Georgi Marinov, Radoslav Mankov, Vihar Belina, Evdokiya Kisov, Hristo Strijkova-Kenderova, Velichka Kantardjiev, Todor Biosensors (Basel) Article Cell-based assays are a valuable tool for examination of virus–host cell interactions and drug discovery processes, allowing for a more physiological setting compared to biochemical assays. Despite the fact that cell-based SPR assays are label-free and thus provide all the associated benefits, they have never been used to study viral growth kinetics and to predict drug antiviral response in cells. In this study, we prove the concept that the cell-based SPR assay can be applied in the kinetic analysis of the early stages of viral infection of cells and the antiviral drug activity in the infected cells. For this purpose, cells immobilized on the SPR slides were infected with human coronavirus HCov-229E and treated with hydroxychloroquine. The SPR response was measured at different time intervals within the early stages of infection. Methyl Thiazolyl Tetrazolium (MTT) assay was used to provide the reference data. We found that the results of the SPR and MTT assays were consistent, and SPR is a reliable tool in investigating virus–host cell interaction and the mechanism of action of viral inhibitors. SPR assay was more sensitive and accurate in the first hours of infection within the first replication cycle, whereas the MTT assay was not so effective. After the second replication cycle, noise was generated by the destruction of the cell layer and by the remnants of dead cells, and masks useful SPR signals. MDPI 2021-07-26 /pmc/articles/PMC8392451/ /pubmed/34436052 http://dx.doi.org/10.3390/bios11080251 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Genova-Kalou, Petia
Dyankov, Georgi
Marinov, Radoslav
Mankov, Vihar
Belina, Evdokiya
Kisov, Hristo
Strijkova-Kenderova, Velichka
Kantardjiev, Todor
SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title_full SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title_fullStr SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title_full_unstemmed SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title_short SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine
title_sort spr-based kinetic analysis of the early stages of infection in cells infected with human coronavirus and treated with hydroxychloroquine
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8392451/
https://www.ncbi.nlm.nih.gov/pubmed/34436052
http://dx.doi.org/10.3390/bios11080251
work_keys_str_mv AT genovakaloupetia sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT dyankovgeorgi sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT marinovradoslav sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT mankovvihar sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT belinaevdokiya sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT kisovhristo sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT strijkovakenderovavelichka sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine
AT kantardjievtodor sprbasedkineticanalysisoftheearlystagesofinfectionincellsinfectedwithhumancoronavirusandtreatedwithhydroxychloroquine