Standardization of the methods and reference materials used to assess virus content in varicella vaccines

Hong, Ji‐Young; Oh, Ho Jung; Lee, Naery; Kim, Dokeun; Yoon, Heui-Seong; Kim, Yeon‐Tae; Chang, Seokkee; Park, Jae‐Hak; Chung, Hai Won

doi:10.1186/s12985-015-0333-1

Cited by 3 publications

(2 citation statements)

References 13 publications

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“…Liquid medium-based plaque assays were performed as previously described ( 39 ), with minor modifications. Briefly, LMH cells were exposed to virus that was diluted in serum-free DMEM for 3 h at 37°C in 5% CO 2 and then washed with phosphate-buffered saline (PBS).…”

Section: Methodsmentioning

confidence: 99%

Genome-Wide Gene Expression Analysis Identifies the Proto-oncogene Tyrosine-Protein Kinase Src as a Crucial Virulence Determinant of Infectious Laryngotracheitis Virus in Chicken Cells

Wang

Han

et al. 2016

J Virol

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Given the side effects of vaccination against infectious laryngotracheitis (ILT), novel strategies for ILT control and therapy are urgently needed. The modulation of host-virus interactions is a promising strategy to combat the virus; however, the interactions between the host and avian ILT herpesvirus (ILTV) are unclear. Using genome-wide transcriptome studies in combination with a bioinformatic analysis, we identified proto-oncogene tyrosine-protein kinase Src (Src) to be an important modulator of ILTV infection. Src controls the virulence of ILTV and is phosphorylated upon ILTV infection. Functional studies revealed that Src prolongs the survival of host cells by increasing the threshold of virus-induced cell death. Therefore, Src is essential for viral replication in vitro and in ovo but is not required for ILTV-induced cell death. Furthermore, our results identify a positive-feedback loop between Src and the tyrosine kinase focal adhesion kinase (FAK), which is necessary for the phosphorylation of either Src or FAK and is required for Src to modulate ILTV infection. To the best of our knowledge, we are the first to identify a key host regulator controlling host-ILTV interactions. We believe that our findings have revealed a new potential therapeutic target for ILT control and therapy. IMPORTANCE Despite the extensive administration of live attenuated vaccines starting from the mid-20th century and the administration of recombinant vaccines in recent years, infectious laryngotracheitis (ILT) outbreaks due to avian ILT herpesvirus (ILTV) occur worldwide annually. Presently, there are no drugs or control strategies that effectively treat ILT. Targeting of host-virus interactions is considered to be a promising strategy for controlling ILTV infections. However, little is known about the mechanisms governing host-ILTV interactions. The results from our study advance our understanding of host-ILTV interactions on a molecular level and provide experimental evidence that it is possible to control ILT via the manipulation of host-virus interactions.

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Section: Methodsmentioning

confidence: 99%

Genome-Wide Gene Expression Analysis Identifies the Proto-oncogene Tyrosine-Protein Kinase Src as a Crucial Virulence Determinant of Infectious Laryngotracheitis Virus in Chicken Cells

Wang

Han

et al. 2016

J Virol

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“…Viral passage was initially implemented in a viral monoclonal manner without interference from other JEV strains; thus, there was no possibility of genome recombination. The parent stocks (F0) were obtained by two rounds of plaque assays ( Hong et al. 2015 ), followed by propagation in BHK cells for 96 h. Confluent monolayer cells in a six-well plate were exposed to 1 ml FBS-free medium fixed with 10 μl viral supernatant (F0) at 37°C for 1 h. Subsequently, the medium was changed, and cells were covered with 4 per cent FBS medium for 96 h for viral replication.…”

Section: Methodsmentioning

confidence: 99%

Viral intra-host evolutionary dynamics revealed via serial passage of Japanese encephalitis virus in vitro

Sun

Liu

et al. 2023

Virus Evolution

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Analyses of viral inter- and intra-host mutations could better guide the prevention and control of infectious diseases. For a long time, studies on viral evolution have focused on viral inter-host variations. Next-generation sequencing has accelerated investigations of viral intra-host diversity. However, the theoretical basis and dynamic characteristics of viral intra-host mutations remain unknown. Here, using serial passages of the SA14-14-2 vaccine strain of Japanese encephalitis virus (JEV) as the in vitro model, the distribution characteristics of 1,788 detected intra-host single nucleotide variations (iSNVs) and their mutated frequencies from 477 deep-sequenced samples were analyzed. Our results revealed that in adaptive (BHK) cells, JEV is under a nearly neutral selection pressure, and both non-synonymous and synonymous mutations represent an S-shaped growth trend over time. A higher positive selection pressure was observed in the non-adaptive (C6/36) cells, and logarithmic growth in non-synonymous iSNVs and linear growth in synonymous iSNVs were observed over time. Moreover, the mutation rates of the NS4B protein and the UTR region of the JEV are significantly different between BHK and C6/36 cells, suggesting that viral selection pressure is regulated by different cellular environments. Additionally, no significant difference was detected in the distribution of mutated frequencies of iSNVs between BHK and C6/36 cells.

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Principles of Vaccine Potency Assays

2018

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Compared with biologics, vaccine potency assays represent a special challenge due to their unique compositions, multivalency, long life cycles and global distribution. Historically, vaccines were released using in vivo potency assays requiring immunization of dozens of animals. Modern vaccines use a variety of newer analytical tools including biochemical, cell-based and immunochemical methods to measure potency. The choice of analytics largely depends on the mechanism of action and ability to ensure lot-to-lot consistency. Live vaccines often require cell-based assays to ensure infectivity, whereas recombinant vaccine potency can be reliably monitored with immunoassays. Several case studies are presented to demonstrate the relationship between mechanism of action and potency assay. A high-level decision tree is presented to assist with assay selection.

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Standardization of the methods and reference materials used to assess virus content in varicella vaccines

Cited by 3 publications

References 13 publications

Genome-Wide Gene Expression Analysis Identifies the Proto-oncogene Tyrosine-Protein Kinase Src as a Crucial Virulence Determinant of Infectious Laryngotracheitis Virus in Chicken Cells

Genome-Wide Gene Expression Analysis Identifies the Proto-oncogene Tyrosine-Protein Kinase Src as a Crucial Virulence Determinant of Infectious Laryngotracheitis Virus in Chicken Cells

Viral intra-host evolutionary dynamics revealed via serial passage of Japanese encephalitis virus in vitro

Principles of Vaccine Potency Assays

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