Enhancing viral vaccine production using engineered knockout vero cell lines – A second look

Hoeksema, Femke; Karpilow, Jon; Luitjens, Alfred; Lagerwerf, Fija M.; Havenga, Menzo; Groothuizen, M.; Gillissen, Gert; Lemckert, Angelique A. C.; Jiang, Baoming; Tripp, Ralph A.; Yallop, Christopher

doi:10.1016/j.vaccine.2018.03.010

Cited by 18 publications

(19 citation statements)

References 24 publications

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“…The antiviral host genes that affect HuNoV replication are not well-understood, but it is known that viruses co-opt host genes to replicate, and host genes are known to be required for virus replication. We took advantage of previous findings of virus-host gene interactions [52,96,97]. Based on genome-wide siRNA screens for several RNA viruses, we examined the top six genes for which KD enhanced virus permissiveness and replication in Vero cells [52].…”

Section: Discussionmentioning

confidence: 99%

Vero Cells as a Mammalian Cell Substrate for Human Norovirus

Todd

Tripp

2020

Viruses

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Human norovirus (HuNoV) is a principal cause of acute gastroenteritis worldwide, particularly in developing countries. Its global prevalence is underscored by more serious morbidity and some mortality in the young (<5 years) and the elderly. To date, there are no licensed vaccines or approved therapeutics for HuNoV, mostly because there are limited cell culture systems and small animal models available. Recently described cell culture systems are not ideal substrates for HuNoV vaccine development because they are not clonal or only support a single strain. In this study, we show Vero cell-based replication of two pandemic GII.4 HuNoV strains and one GII.3 strain and confirm exosome-mediated HuNoV infection in Vero cells. Lastly, we show that trypsin addition to virus cultures or disruption of Vero cell host genes can modestly increase HuNoV replication. These data provide support for Vero cells as a cell culture model for HuNoV.

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Section: Discussionmentioning

confidence: 99%

Vero Cells as a Mammalian Cell Substrate for Human Norovirus

Todd

Tripp

2020

Viruses

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“…identified several gene knockdowns that drastically increased yields of multiple PV, enterovirus and rotavirus strains. However, these effects on viral replication were not recapitulated on follow‐up . As the reasons for these discrepant results remain unclear, challenges evidently remain in engineering cell lines that support greater viral yields for vaccine deployment.…”

Section: Challenges and Strategies In Targeting Multifunctional Host mentioning

confidence: 99%

Harnessing host–virus evolution in antiviral therapy and immunotherapy

Heaton

2019

Clin & Trans Imm

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Pathogen resistance and development costs are major challenges in current approaches to antiviral therapy. The high error rate of RNA synthesis and reverse‐transcription confers genome plasticity, enabling the remarkable adaptability of RNA viruses to antiviral intervention. However, this property is coupled to fundamental constraints including limits on the size of information available to manipulate complex hosts into supporting viral replication. Accordingly, RNA viruses employ various means to extract maximum utility from their informationally limited genomes that, correspondingly, may be leveraged for effective host‐oriented therapies. Host‐oriented approaches are becoming increasingly feasible because of increased availability of bioactive compounds and recent advances in immunotherapy and precision medicine, particularly genome editing, targeted delivery methods and RNAi. In turn, one driving force behind these innovations is the increasingly detailed understanding of evolutionarily diverse host–virus interactions, which is the key concern of an emerging field, neo‐virology. This review examines biotechnological solutions to disease and other sustainability issues of our time that leverage the properties of RNA and DNA viruses as developed through co‐evolution with their hosts.

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“…To overcome this limitation, one option is to manipulate the expression level of host cell factors (HCFs) relevant for virus replication in order to enhance virus yield. However, to identify promising HCF candidates costly and time-consuming experimental screening and validation studies are required, as shown recently for poliovirus [2,3]. This motivates the development of suitable computational tools to predict the impact of genetic modifications in face of the inevitable cellular heterogeneity on the overall product yield.…”

Section: Introductionmentioning

confidence: 99%

Model-based approach for predicting the impact of genetic modifications on product yield in biopharmaceutical manufacturing—Application to influenza vaccine production

et al. 2020

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A large group of biopharmaceuticals is produced in cell lines. The yield of such products can be increased by genetic engineering of the corresponding cell lines. The prediction of promising genetic modifications by mathematical modeling is a valuable tool to facilitate experimental screening. Besides information on the intracellular kinetics and genetic modifications the mathematical model has to account for ubiquitous cell-to-cell variability. In this contribution, we establish a novel model-based methodology for influenza vaccine production in cell lines with overexpressed genes. The manipulation of the expression level of genes coding for host cell factors relevant for virus replication is achieved by lentiviral transduction. Since lentiviral transduction causes increased cell-to-cell variability due to different copy numbers and integration sites of the gene constructs we use a population balance modeling approach to account for this heterogeneity in terms of intracellular viral components and distributed kinetic parameters. The latter are estimated from experimental data of intracellular viral RNA levels and virus titers of infection experiments using cells overexpressing a single host cell gene. For experiments with cells overexpressing multiple host cell genes, only final virus titers were measured and thus, no direct estimation of the parameter distributions was possible. Instead, we evaluate four different computational strategies to infer these from single gene parameter sets. Finally, the best computational strategy is used to predict the most promising candidates for future modifications that show the highest potential for an increased virus yield in a combinatorial study. As expected, there is a trend to higher yields the more modifications are included.

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Enhancing viral vaccine production using engineered knockout vero cell lines – A second look

Cited by 18 publications

References 24 publications

Vero Cells as a Mammalian Cell Substrate for Human Norovirus

Vero Cells as a Mammalian Cell Substrate for Human Norovirus

Harnessing host–virus evolution in antiviral therapy and immunotherapy

Model-based approach for predicting the impact of genetic modifications on product yield in biopharmaceutical manufacturing—Application to influenza vaccine production

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