Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2
Introduction. The continued epidemic relevance of SARS-CoV-2, even after the end of the associated COVID-19 pandemic in 2023, necessitates further study of the interaction of this virus with the first line of cellular defense, neutrophils. The aim of the study was to determine the enzymatic activ...
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Central Research Institute for Epidemiology
2025-05-01
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Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
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Online Access: | https://microbiol.crie.ru/jour/article/viewFile/18825/1589 |
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author | Svetlana A. Abramova Irina N. Lyapun Elena I. Drobot Natalia V. Krylova Olga V. Iunikhina Valeria A. Lubova Evgeniy K. Merlov Iurii A. Belov Larisa M. Somova Mikhail Yu. Shchelkanov |
author_facet | Svetlana A. Abramova Irina N. Lyapun Elena I. Drobot Natalia V. Krylova Olga V. Iunikhina Valeria A. Lubova Evgeniy K. Merlov Iurii A. Belov Larisa M. Somova Mikhail Yu. Shchelkanov |
author_sort | Svetlana A. Abramova |
collection | DOAJ |
description | Introduction. The continued epidemic relevance of SARS-CoV-2, even after the end of the associated COVID-19 pandemic in 2023, necessitates further study of the interaction of this virus with the first line of cellular defense, neutrophils.
The aim of the study was to determine the enzymatic activity of peripheral blood leukocytes of Syrian hamsters (Mesocricetus auratus) in the dynamics of ex vivo SARS-CoV-2 infection, which characterizes the microbicidal potential of innate immunity cells.
Materials and methods. The SARS-CoV-2/Vladivostok/R-8726/2021 strain was used at infectious doses of 3 lg (TCID50/mL) and 2 lg (TCID50/mL) (TCID50 is the 50% tissue cytopathic dose for the Vero E6 cell line); the contact time of the infecting virus-containing liquid with the cell culture was 1 h. The number of viable cells in the adherent leukocyte culture was counted using an inverted microscope equipped with a digital camera and MCView program. The specific (per 1 viable cell) activities of adenosine triphosphatase (ATPase), 5′-nucleotidase (adenosine monophosphatase, AMPase), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), myeloperoxidase (MPO) and cytochrome oxidase (CCO) were determined spectrophotometrically after incubation with specific substrates of infected and uninfected cell cultures 1, 16, 24, 48 h after virus inoculation.
Results. The enzymatic activity of leukocytes 1 h after virus inoculation, compared to uninfected leukocytes, was as follows: specific activity of ATPase, MPO was decreased, activity of AMPase, LDH, SDH was increased; 16 h after virus inoculation, activity of MPO was decreased, activity of AMPase, LDH, SDH was increased, activity of ATPase and CCO was at the initial level, i.e. approximately at the level of the uninfected control; 24 h after virus inoculation, AMPase activity was decreased, ATPase activity was increased, LDH, SDH, MPO, CCO activity was at the initial level; 48 h after virus inoculation, ATPase, LDH, SDH, MPO, CCO activity was increased, AMPase activity was at the initial level. Changes in enzymatic activity depend on the infecting dose and correlate with virus accumulation in the culture medium.
Conclusion. The revealed dynamics of enzymatic activity in the primary culture of adherent leukocytes ex vivo infected with SARS-CoV-2 indicates a decrease in the microbicidal potential of cells of innate immunity in the course of this infection. |
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publishDate | 2025-05-01 |
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spelling | doaj-art-a0b38cfe5e2c41c4a14d918b9bbadec52025-07-03T19:50:55ZrusCentral Research Institute for EpidemiologyЖурнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132025-05-01102216817810.36233/0372-9311-6492818Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2Svetlana A. Abramova0https://orcid.org/0000-0002-2428-3186Irina N. Lyapun1https://orcid.org/0000-0002-5290-3864Elena I. Drobot2https://orcid.org/0000-0001-7672-1582Natalia V. Krylova3https://orcid.org/0000-0002-9048-6803Olga V. Iunikhina4https://orcid.org/0000-0002-6723-582XValeria A. Lubova5https://orcid.org/0000-0002-4290-6164Evgeniy K. Merlov6https://orcid.org/0000-0003-1515-3221Iurii A. Belov7https://orcid.org/0000-0001-8313-5610Larisa M. Somova8https://orcid.org/0000-0003-2023-1503Mikhail Yu. Shchelkanov9https://orcid.org/0000-0001-8610-7623Somov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologySomov Institute of Epidemiology and MicrobiologyIntroduction. The continued epidemic relevance of SARS-CoV-2, even after the end of the associated COVID-19 pandemic in 2023, necessitates further study of the interaction of this virus with the first line of cellular defense, neutrophils. The aim of the study was to determine the enzymatic activity of peripheral blood leukocytes of Syrian hamsters (Mesocricetus auratus) in the dynamics of ex vivo SARS-CoV-2 infection, which characterizes the microbicidal potential of innate immunity cells. Materials and methods. The SARS-CoV-2/Vladivostok/R-8726/2021 strain was used at infectious doses of 3 lg (TCID50/mL) and 2 lg (TCID50/mL) (TCID50 is the 50% tissue cytopathic dose for the Vero E6 cell line); the contact time of the infecting virus-containing liquid with the cell culture was 1 h. The number of viable cells in the adherent leukocyte culture was counted using an inverted microscope equipped with a digital camera and MCView program. The specific (per 1 viable cell) activities of adenosine triphosphatase (ATPase), 5′-nucleotidase (adenosine monophosphatase, AMPase), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), myeloperoxidase (MPO) and cytochrome oxidase (CCO) were determined spectrophotometrically after incubation with specific substrates of infected and uninfected cell cultures 1, 16, 24, 48 h after virus inoculation. Results. The enzymatic activity of leukocytes 1 h after virus inoculation, compared to uninfected leukocytes, was as follows: specific activity of ATPase, MPO was decreased, activity of AMPase, LDH, SDH was increased; 16 h after virus inoculation, activity of MPO was decreased, activity of AMPase, LDH, SDH was increased, activity of ATPase and CCO was at the initial level, i.e. approximately at the level of the uninfected control; 24 h after virus inoculation, AMPase activity was decreased, ATPase activity was increased, LDH, SDH, MPO, CCO activity was at the initial level; 48 h after virus inoculation, ATPase, LDH, SDH, MPO, CCO activity was increased, AMPase activity was at the initial level. Changes in enzymatic activity depend on the infecting dose and correlate with virus accumulation in the culture medium. Conclusion. The revealed dynamics of enzymatic activity in the primary culture of adherent leukocytes ex vivo infected with SARS-CoV-2 indicates a decrease in the microbicidal potential of cells of innate immunity in the course of this infection.https://microbiol.crie.ru/jour/article/viewFile/18825/1589sars-cov-2leukocytesneutrophilsatpase5′-nucleotidaselactate dehydrogenasesuccinate dehydrogenasemyeloperoxidasecytochrome oxidasemicrobicidal potential |
spellingShingle | Svetlana A. Abramova Irina N. Lyapun Elena I. Drobot Natalia V. Krylova Olga V. Iunikhina Valeria A. Lubova Evgeniy K. Merlov Iurii A. Belov Larisa M. Somova Mikhail Yu. Shchelkanov Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 Журнал микробиологии, эпидемиологии и иммунобиологии sars-cov-2 leukocytes neutrophils atpase 5′-nucleotidase lactate dehydrogenase succinate dehydrogenase myeloperoxidase cytochrome oxidase microbicidal potential |
title | Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 |
title_full | Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 |
title_fullStr | Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 |
title_full_unstemmed | Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 |
title_short | Dynamics of enzymatic activity in primary culture of Syrian hamster adherent leukocytes <i>ex vivo</i> infected with SARS-CoV-2 |
title_sort | dynamics of enzymatic activity in primary culture of syrian hamster adherent leukocytes i ex vivo i infected with sars cov 2 |
topic | sars-cov-2 leukocytes neutrophils atpase 5′-nucleotidase lactate dehydrogenase succinate dehydrogenase myeloperoxidase cytochrome oxidase microbicidal potential |
url | https://microbiol.crie.ru/jour/article/viewFile/18825/1589 |
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