Kinetic model for adherent Vero cell growth and poliovirus production in batch bioreactors

Jiang, Yang; Welle, Joyce E. van der; Rubingh, Olaf; Eikenhorst, Gerco van; Bakker, Wilfried A.M.; Thomassen, Yvonne E.

doi:10.1016/j.procbio.2019.03.010

Cited by 14 publications

(7 citation statements)

References 28 publications

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“…We chose the target cell proliferation and death rate constants representative of Vero cells, typically used for studying SARS-CoV-2 infections in vitro . These have been estimated to be 0.77 d -1 and 0.22 d -1 [ 82 , 83 ]. The infected cell death rate and the virion clearance rate constants have been chosen to apply to SARS-CoV-2 infections.…”

Section: Methodsmentioning

confidence: 99%

Targeting TMPRSS2 and Cathepsin B/L together may be synergistic against SARS-CoV-2 infection

2020

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The entry of SARS-CoV-2 into target cells requires the activation of its surface spike protein, S, by host proteases. The host serine protease TMPRSS2 and cysteine proteases Cathepsin B/L can activate S, making two independent entry pathways accessible to SARS-CoV-2. Blocking the proteases prevents SARS-CoV-2 entry in vitro. This blockade may be achieved in vivo through ‘repurposing’ drugs, a potential treatment option for COVID-19 that is now in clinical trials. Here, we found, surprisingly, that drugs targeting the two pathways, although independent, could display strong synergy in blocking virus entry. We predicted this synergy first using a mathematical model of SARS-CoV-2 entry and dynamics in vitro. The model considered the two pathways explicitly, let the entry efficiency through a pathway depend on the corresponding protease expression level, which varied across cells, and let inhibitors compromise the efficiency in a dose-dependent manner. The synergy predicted was novel and arose from effects of the drugs at both the single cell and the cell population levels. Validating our predictions, available in vitro data on SARS-CoV-2 and SARS-CoV entry displayed this synergy. Further, analysing the data using our model, we estimated the relative usage of the two pathways and found it to vary widely across cell lines, suggesting that targeting both pathways in vivo may be important and synergistic given the broad tissue tropism of SARS-CoV-2. Our findings provide insights into SARS-CoV-2 entry into target cells and may help improve the deployability of drug combinations targeting host proteases required for the entry.

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Section: Methodsmentioning

confidence: 99%

Targeting TMPRSS2 and Cathepsin B/L together may be synergistic against SARS-CoV-2 infection

2020

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show abstract

“…We chose the target cell proliferation and death rate constants representative of Vero cells, typically used for studying SARS-CoV-2 infections in vitro. These have been estimated to be 0.77 d -1 and 0.22 d -1 [82,83]. The infected cell death rate and the virion clearance rate constants have been chosen to apply to SARS-CoV-2 infections.…”

Section: Model Calculations and Parametersmentioning

confidence: 99%

Targeting TMPRSS2 and Cathepsin B/L Together May Be Synergistic Against SARS-CoV-2 Infection

Padmanabhan¹,

Desikan²,

Dixit

2020

Preprint

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The entry of SARS-CoV-2 into target cells requires the activation of its surface spike protein, S, by host proteases. The host serine protease TMPRSS2 and cysteine proteases Cathepsin B/L can activate S, making two independent entry pathways accessible to SARS-CoV-2. Blocking the proteases prevents SARS-CoV-2 entry in vitro. This blockade may be achieved in vivo through ‘repurposing’ drugs, a potential treatment option for COVID-19 that is now in clinical trials. Here, we found, surprisingly, that drugs targeting the two pathways, although independent, could display strong synergy in blocking virus entry. We predicted this synergy first using a mathematical model of SARS-CoV-2 entry and dynamics in vitro. The model considered the two pathways explicitly, let the entry efficiency through a pathway depend on the corresponding protease expression level, which varied across cells, and let inhibitors compromise the efficiency in a dose-dependent manner. The synergy predicted was novel and arose from effects of the drugs at both the single cell and the cell population levels. Validating our predictions, available in vitro data on SARS-CoV-2 and SARS-CoV entry displayed this synergy. Further, analysing the data using our model, we estimated the relative usage of the two pathways and found it to vary widely across cell lines, suggesting that targeting both pathways in vivo may be important and synergistic given the broad tissue tropism of SARS-CoV-2. Our findings provide insights into SARS-CoV-2 entry into target cells and may help improve the deployability of drug combinations targeting host proteases required for the entry.

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“…The models may further account for spread owing to fluid flows or employ rule‐based cellular automata to model the coupling of virus growth with the spatial transport of virus particles (Akpinar et al, 2016; Anekal et al, 2009; Mitchell et al, 2011). In other scenarios of vaccine production, the microcarrier culture of Vero cells may be closer to well‐mixed systems (Jiang et al, 2019; Souza et al, 2009; Sugawara et al, 2002), so the present simpler models might in such cases provide a starting point for modeling.…”

Section: Discussionmentioning

confidence: 99%

Kinetics of Asian and African Zika virus lineages over single‐cycle and multi‐cycle growth in culture: Gene expression, cell killing, virus production, and mathematical modeling

Shi

Yin

2021

Biotech & Bioengineering

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Since 2014, an Asian lineage of Zika virus has caused outbreaks, and it has been associated with neurological disorders in adults and congenital defects in newborns.The resulting threat of the Zika virus to human health has prompted the development of new vaccines, which have yet to be approved for human use. Vaccines based on the attenuated or chemically inactivated virus will require large-scale production of the intact virus to meet potential global demands. Intact viruses are produced by infecting cultures of susceptible cells, a dynamic process that spans from hours to days and has yet to be optimized. Here, we infected Vero cells adhesively cultured in well-plates with two Zika virus strains: a recently isolated strain from the Asian lineage, and a cell-culture-adapted strain from the African lineage. At different time points post-infection, virus particles in the supernatant were quantified; further, microscopy images were used to quantify cell density and the proportion of cells expressing viral protein. These measurements were performed across multiple replicate samples of one-step infections every four hours over 60 h and for multi-step infections every four to 24 h over 144 h, generating a rich data set. For each set of data, mathematical models were developed to estimate parameters associated with cell infection and virus production. The African-lineage strain was found to produce a 14-fold higher yield than the Asian-lineage strain in one-step growth and a sevenfold higher titer in multi-step growth, suggesting a benefit of cell-culture adaptation for developing a vaccine strain. We found that image-based measurements were critical for discriminating among different models, and different parameters for the two strains could account for the experimentally observed differences. An exponential-distributed delay model performed best in accounting for multi-step infection of the Asian strain, and it highlighted the significant sensitivity of virus titer to the rate of viral degradation, with implications for optimization of vaccine production. More broadly, this study highlights how imagebased measurements can contribute to the discrimination of virus-culture models for the optimal production of inactivated and attenuated whole-virus vaccines.

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Kinetic model for adherent Vero cell growth and poliovirus production in batch bioreactors

Abstract: Graphical abstract

Cited by 14 publications

References 28 publications

Targeting TMPRSS2 and Cathepsin B/L together may be synergistic against SARS-CoV-2 infection

Targeting TMPRSS2 and Cathepsin B/L together may be synergistic against SARS-CoV-2 infection

Targeting TMPRSS2 and Cathepsin B/L Together May Be Synergistic Against SARS-CoV-2 Infection

Kinetics of Asian and African Zika virus lineages over single‐cycle and multi‐cycle growth in culture: Gene expression, cell killing, virus production, and mathematical modeling

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