Ultrastructural organization and virion production in Vero E6 cell line under infection with the Delta and Omicron SARS-CoV-2 genovariants

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Abstract

BACKGROUND: The emergence of the coronavirus (SARS-CoV-2) caused a pandemic that led to an increase in morbidity and mortality among infected individuals. The SARS-CoV-2 variants circulating during that period, including Delta, were characterized by a broad spectrum of pathological effects at both the tissue and cellular levels. In 2025, the Omicron variant predominates, producing milder forms of respiratory conditions.

Among the key research questions related to SARS-CoV-2, considerable attention has been devoted to its pathogenicity, which is characterized by high virulence and the ability to induce systemic pathological processes.

Transmission electron microscopy is used to visualize virus–cell receptor interactions and to study the destructive impact of the pathogen on host cells. The findings of such studies form the basis for understanding the course and pathogenesis of the disease and contribute to vaccine development.

AIM: The work aimed to investigate SARS-CoV-2 virion reproduction and the trends of cytopathic effects in Vero E6 cell culture infected with Delta and Omicron variants.

METHODS: Ultrastructural analysis was performed on Vero E6 cell cultures infected with the Delta and Omicron SARS-CoV-2 variants. Infected cell cultures were fixed at three time points: 6, 18, and 24 hours after viral inoculation.

RESULTS: Most samples in both groups (Delta and Omicron) retained near-normal morphology 6 hours post-inoculation and exhibited numerous transport vesicles. At 18 hours, Delta and Omicron groups showed similar morphological changes, including vacuolization of the endoplasmic reticulum and Golgi complex. The Omicron group demonstrated altered mitochondrial morphology. Marked destructive changes were observed 24 hours post-inoculation. Both groups displayed cytoplasmic condensation, extensive organelle vacuolization, and numerous vesicles containing viral particles. Analysis of virion reproduction trends revealed an increase in viral particle numbers at 18 hours, followed by a decline in viral virion replication by 24 hours.

CONCLUSION: The study suggests similar mechanisms of cytopathic pattern formation in Vero E6 cells infected with the Delta and Omicron SARS-CoV-2 variants, along with active virion reproduction in cell culture between 6 and 18 hours post-infection.

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About the authors

Ekaterina V. Spiridonova

State Research Center of Virology and Biotechnology “Vector”

Email: spiridonova_ev@vector.nsc.ru
ORCID iD: 0009-0006-8655-6713
Russian Federation, Koltsovo

Ksenia F. Emtsova

State Research Center of Virology and Biotechnology “Vector”

Email: emtsova_kf@vector.nsc.ru
ORCID iD: 0009-0003-5165-5357
Russian Federation, Koltsovo

Vladimir V. Omigov

State Research Center of Virology and Biotechnology “Vector”

Email: omigov_vv@vector.nsc.ru
ORCID iD: 0000-0002-2028-6099
SPIN-code: 8454-9741

MD, Cand. Sci. (Medicine)

Russian Federation, Koltsovo

Oleg S. Taranov

State Research Center of Virology and Biotechnology “Vector”

Email: taranov@vector.nsc.ru
ORCID iD: 0000-0002-6746-8092
Russian Federation, Koltsovo

Anastasia A. Moiseeva

State Research Center of Virology and Biotechnology “Vector”

Email: chalaya_aa@vector.nsc.ru
ORCID iD: 0000-0001-7048-2357
Russian Federation, Koltsovo

Elena I. Danilenko

State Research Center of Virology and Biotechnology “Vector”

Author for correspondence.
Email: danilenko_ei@vector.nsc.ru
ORCID iD: 0009-0007-8106-7037
SPIN-code: 8890-7214
Russian Federation, Koltsovo

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Supplementary files

Supplementary Files
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1. JATS XML
2. Fig. 1. Fragments of two Vero E6 cells: dashed arrows indicate multiple profiles of granular endoplasmic reticulum; solid arrows indicate multiple profiles of the Golgi apparatus; phagosomes (F) and nuclei (N) are visible in the hyaloplasm. Electron micrograph; scale bar, 1 µm.

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3. Fig. 2. Fragment of the cytoplasm of a Vero E6 cell: M, numerous mitochondria of typical morphology with clearly delineated lamellar cristae. Electron micrograph; scale bar, 1 µm.

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4. Fig. 3. Ultrastructural alterations in Vero E6 cell culture infected with the Delta SARS-CoV-2 variant: a–c, 6 hours post-inoculation; d, e, 18 hours post-inoculation; f, 24 hours post-inoculation. а , intercellular space; dashed arrow — SARS-CoV-2 particles in the process of adhesion; solid arrow — elements of inoculation; b, peripheral cytoplasmic region; solid arrow — large vesicles containing viral particles; dashed arrow — moderately vacuolized structures of the Golgi complex (GC), N — nucleus; c, peripheral cytoplasmic region; arrows indicate mitochondria with a light, finely granular matrix and large vesicles of the endoplasmic reticulum (V); d, pronounced cellular destruction with cytoplasmic matrix condensation and vacuolization (arrow), N — nucleus; e, perinuclear area (N); arrows indicate ordered accumulations of replicated virions on the inner surface of transport vesicle membranes; f, cytoplasmic area; arrows indicate vesicles containing replicated viral particles; g, quantitative changes in viral particles in Vero E6 cells 6, 18, and 24 hours post-inoculation with the Delta SARS-CoV-2 variant. Scale bars: a–c, e, 500 nm; d, 2 µm; f, 200 nm.

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5. Fig. 4. Ultrastructural alterations in Vero E6 cell culture infected with the Omicron SARS-CoV-2 variant: a–c, 6 hours post-inoculation; d, e, 18 hours post-inoculation; f, g, 24 hours post-inoculation. а, outer cell membrane with an attached virion (solid arrow) and two profiles of inoculating virions (dashed arrows); b, cytoplasmic region near the nucleus (N); arrows indicate swollen large mitochondria; c, cell fragment showing signs of high functional activity of cytoplasmic organelles, as evidenced by pronounced vacuolization of the Golgi complex (GC); arrows indicate Golgi membrane elements containing newly formed virions; d, perinuclear region with large vesicles (arrows) containing virions attached to the membrane; e, cell in a state of destruction; fragments of membranous components are visible against the optically light hyaloplasm; arrows indicate swollen mitochondria; f, two necrotized cells characterized by high optical density of the cytoplasm and extensive vacuolization of organelles; N, nucleus; g, cytoplasmic fragment with swollen mitochondria (arrows); h, quantitative changes in viral particles in Vero E6 cells 6, 18, and 24 hours post-inoculation with the Omicron SARS-CoV-2 variant. Scale bars: a, 200 nm; b, c, e, g, 1 µm; d, 500 nm; f, 2 µm.

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