Production of high titer attenuated poliovirus strains on the serum-free PER.C6® cell culture platform for the generation of safe and affordable next generation IPV

Sanders, Barbara P.; Oakes, Isabel de los Rios; Hoek, Vladimir van; Liu, Ying; Marissen, Wilfred Ε.; Minor, Philip D.; Wimmer, Eckard; Schuitemaker, Hanneke; Custers, Jerome; Macadam, Andrew; Cello, Jerónimo; Edo-Matas, Diana

doi:10.1016/j.vaccine.2015.10.091

Cited by 17 publications

(14 citation statements)

References 31 publications

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“…† Sum of three independent analyses. hyper-attenuated strains for vaccine production has been shown 18 and was confirmed for the monitoring of PV neutralizing antibody titers in the QC of immunoglobulin, another essential medicinal, in this work. Using standard virological practices it was possible to produce S19 stocks with infectivity titers that were suitable for QC release testing, the only special requirement for the handling of these strains stemming from their genetically modified nature, i.e., laboratories licensed for work with GMOs might be required.…”

Section: Discussionsupporting

confidence: 68%

“…Replication incompetent pseudoviruses or hyper‐attenuated PV strains have been developed and their use in vaccine production, immunogenicity studies, immunotherapy, monitoring of the environment, or neutralizing antibody titers has been proposed. The feasibility to use such hyper‐attenuated strains for vaccine production has been shown and was confirmed for the monitoring of PV neutralizing antibody titers in the QC of immunoglobulin, another essential medicinal, in this work.…”

Section: Discussionsupporting

confidence: 57%

“…It is likely that the strains will also prove suitable for other activities requiring live virus such as seroprevalence studies, vaccine potency assays, and validation of cell‐sensitivity. As feasibility for their use in vaccine production has previously been shown, these hyper‐attenuated strains now need to be recognized as safe alternatives. As long as good virological practice is maintained, based on a seed lot system with stocks characterized using the sensitive assays described here, S19 strains should be exempt from full GAPIII containment requirements in order to allow for their establishment in the activities that will need to continue in the future, which will in fact contribute to final containment and aid in achieving and maintaining a PV free world.…”

Section: Discussionmentioning

confidence: 99%

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Continued use of poliovirus after eradication: hyper‐attenuated strains as a safe alternative for release testing of human immunoglobulins

Farcet¹,

Modrof²,

Rabel³

et al. 2018

Transfusion

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BACKGROUND Wild‐type poliovirus may be eradicated soon and under WHO GAPIII guidance, laboratory use will be discontinued or subject to strict containment. Per US Code of Federal Regulations, however, immunoglobulin lot release testing will still require use of replicating poliovirus. The suitability of S19 hyper‐attenuated and apathogenic poliovirus strains as alternatives to the currently used wild‐type virus in such a release assay was investigated. STUDY DESIGN AND METHODS S19 poliovirus strains were propagated in a commercial setting using good virological practices and maintenance of the S19 hyper‐attenuated genotype was confirmed by massively parallel sequencing. RESULTS The attenuated phenotype of the produced S19 stocks was confirmed in a highly sensitive mouse‐model. Equivalency in performance was seen in the lot release assay for the S19 and wild‐type polioviruses. CONCLUSION The deployment of such hyper‐attenuated and thoroughly characterized S19 stocks in these and other essential activities might reconcile final containment measures with continued safe use of poliovirus.

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Section: Discussionsupporting

confidence: 68%

Section: Discussionsupporting

confidence: 57%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Continued use of poliovirus after eradication: hyper‐attenuated strains as a safe alternative for release testing of human immunoglobulins

Farcet¹,

Modrof²,

Rabel³

et al. 2018

Transfusion

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“…Virus production during vaccine manufacture with high yields can significantly decrease costs of goods. We have previously demonstrated that the use of the PER.C6 platform increases volumetric productivity of infection harvests as compared to the Vero-based platform for cIPV strains [ 60 ] and for the Sabin strains [ 61 ]. Use of the highly productive PER.C6 platform for propagation of the CAVA vaccine strains resulted high infectious titer yields (9.4–9.9 Log 10 TCID 50 /ml), comparable to those reached with the Sabin strains (9.2–9.9 Log 10 TCID 50 /ml), which demonstrates potential for significantly reducing cost of goods and consequent vaccine pricing required for global roll-out of an affordable IPV.…”

Section: Discussionmentioning

confidence: 99%

Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine

et al. 2016

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The poliovirus vaccine field is moving towards novel vaccination strategies. Withdrawal of the Oral Poliovirus Vaccine and implementation of the conventional Inactivated Poliovirus Vaccine (cIPV) is imminent. Moreover, replacement of the virulent poliovirus strains currently used for cIPV with attenuated strains is preferred. We generated Cold-Adapted Viral Attenuation (CAVA) poliovirus strains by serial passage at low temperature and subsequent genetic engineering, which contain the capsid sequences of cIPV strains combined with a set of mutations identified during cold-adaptation. These viruses displayed a highly temperature sensitive phenotype with no signs of productive infection at 37°C as visualized by electron microscopy. Furthermore, decreases in infectious titers, viral RNA, and protein levels were measured during infection at 37°C, suggesting a block in the viral replication cycle at RNA replication, protein translation, or earlier. However, at 30°C, they could be propagated to high titers (9.4–9.9 Log10TCID50/ml) on the PER.C6 cell culture platform. We identified 14 mutations in the IRES and non-structural regions, which in combination induced the temperature sensitive phenotype, also when transferred to the genomes of other wild-type and attenuated polioviruses. The temperature sensitivity translated to complete absence of neurovirulence in CD155 transgenic mice. Attenuation was also confirmed after extended in vitro passage at small scale using conditions (MOI, cell density, temperature) anticipated for vaccine production. The inability of CAVA strains to replicate at 37°C makes reversion to a neurovirulent phenotype in vivo highly unlikely, therefore, these strains can be considered safe for the manufacture of IPV. The CAVA strains were immunogenic in the Wistar rat potency model for cIPV, inducing high neutralizing antibody titers in a dose-dependent manner in response to D-antigen doses used for cIPV. In combination with the highly productive PER.C6 cell culture platform, the stably attenuated CAVA strains may serve as an attractive low-cost and (bio)safe option for the production of a novel next generation IPV.

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“…Beside these advantages for basic research, some Ad t cell lines such as PER.C6 and AGE.CR.pIX provide a potential platform for the manufacturing of viral vectors and viral vaccines already used in clinical trials (Koudstaal et al, 2009;Lohr et al, 2009Lohr et al, , 2012Pau et al, 2001;Subramanian et al, 2007). These cell lines were shown to be a good substrate for a wide spectrum of viruses, such as poliovirus (Sanders et al, 2015), modified vaccinia Ankara virus (Jordan et al, 2009b) and West Nile virus (Samina et al, 2007). In addition, several Ad t cell lines can be cultivated in suspension allowing a better scalability and flexibility in virus production processes …”

Section: Adenoviral Protein E1a Inhibits the Antiviral Responsementioning

confidence: 99%

Impaired antiviral response of adenovirus-transformed cell lines supports virus replication

Bachmann

Breitwieser

Lipps

et al. 2016

Journal of General Virology

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Activation of the innate immune response represents one of the most important cellular mechanisms to limit virus replication and spread in cell culture. Here, we examined the effect of adenoviral gene expression on the antiviral response in adenovirus-transformed cell lines; HEK293, HEK293SF and AGE1.HN. We demonstrate that the expression of the early region protein 1A in these cell lines impairs their ability to activate antiviral genes by the IFN pathway. This property may help in the isolation of newly emerging viruses and the propagation of interferon-sensitive virus strains. Received 30 October 2015 Accepted 4 December 2015The IFN pathway of the innate immune response plays an important role in the control of viral infections. Once a cell detects viral components via its intracellular pathogen recognition receptors, type I IFNs (IFN-a/b) are synthesized and secreted, which act in an auto-and paracrine fashion and lead to the expression of IFN-stimulated genes (ISGs). Some of these ISGs, such as the myxovirus resistance A protein (MxA), possess direct antiviral activities (Haller et al., 2009). Viruses have developed efficient strategies to inhibit the IFN response (García-Sastre, 2011) and escape these antiviral mechanisms. Still, suppression of the antiviral response by chemical inhibitors, expression of viral antagonists or knockdown of antiviral proteins can improve permissiveness for emerging virus strains and increase yields in vaccine production (McSharry et al., 2015;Stewart et al., 2014;van Wielink et al., 2011;Young et al., 2003;Zhu et al., 2014). Inhibition of the IFN pathway can also support the propagation of IFN-sensitive viruses, like the influenza A virus (IAV) deletion strain that lacks its non-structural protein 1 (delNS1). The delNS1 strain is attenuated in IFN-competent cells and hence, is considered as a live vaccine for both animals (Richt & García-Sastre, 2009;Wang et al., 2008) and humans (Mössler et al., 2013;Wressnigg et al., 2009). So far, only a few cell lines, such as Vero cells, which are unable to produce IFN, support the propagation of IFN-sensitive viruses (Barrett et al., 2009;Emeny & Morgan, 1979;Stewart et al., 2014). However, not all viruses replicate in these African green monkey cells and some properties of the virus, such as the glycan structures of viral surface proteins, can be altered by propagation in cells that do not originate from natural host organisms (Dumont et al., 2015;Genzel et al., 2012; Schwarzer et al., 2009). Thus, there is a need to provide flexible host cell systems for the propagation of IFN-sensitive viruses.In this study, we examined the effect of adenovirus early region protein 1A (E1A) and 1B (E1B) expression on the antiviral response and permissiveness for IFN-sensitive viruses in the adenovirus-transformed (Ad t ) cell lines, AGE1.HN (Niklas et al., 2011), HEK293 and HEK293SF [a derivative of HEK293 cells growing in serum-free (SF) medium and adapted to the proliferation in suspension (Cô té et al., 1998)]. IAV efficiently replicates in adeno...

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Production of high titer attenuated poliovirus strains on the serum-free PER.C6® cell culture platform for the generation of safe and affordable next generation IPV

Cited by 17 publications

References 31 publications

Continued use of poliovirus after eradication: hyper‐attenuated strains as a safe alternative for release testing of human immunoglobulins

Continued use of poliovirus after eradication: hyper‐attenuated strains as a safe alternative for release testing of human immunoglobulins

Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine

Impaired antiviral response of adenovirus-transformed cell lines supports virus replication

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