Principles of selective inactivation of viral genome. VI. Inactivation of the infectivity of the influenza virus by the action of β-propiolactone

Budowsky, E.I.; Friedman, Edgar; Железнова, Н. В.; Noskov, F. S.

doi:10.1016/0264-410x(91)90125-p

Cited by 58 publications

(47 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A. laidlawii remained viable for 2 h of incubation with 0.1% BPL; this result is consistent with the ability of this species to resist inactivation by BPL described previously (30). In a majority of the protocols used for production of inactivated virus vaccines (2,9,17,25,27,29,33,42,51,53), the concentration of BPL and formaldehyde exceeds the critical concentration (0.1% to 0.2%) required for complete inactivation of mycoplasmas. Our data showed that these conditions resulted in complete inactivation of many of the mycoplasmas evaluated.…”

Section: Discussionsupporting

confidence: 68%

“…BPL and formaldehyde are commonly used at concentrations of 0.05 to 0.2% as primary chemical agents for inactivation of vaccine viruses in cell-and eggderived harvests. The inactivating effect of these reagents is based on their chemical reaction with nucleophilic groups of viral proteins and nucleic acids that results in irreversible changes in the structure of virions and the loss of viral infectivity (8,9). The concentrations of chemicals, time, and temperature required for efficient virus inactivation may vary significantly depending on the type of virus and the protocol used by the vaccine manufacturer.…”

Section: Resultsmentioning

confidence: 99%

“…To assess the contribution of each chemical agent used in the mycoplasma inactivation process, the agents were tested in separate experiments at concentrations that are commonly used or represent "worst-case scenarios" for flu vaccine production. The first group of reagents included formaldehyde and BPL, which are commonly used for inactivation of vaccine viruses via chemical reaction with viral capsid proteins and nucleic acids (3,8,9). The second group consisted of surfactants, including cetyltrimethylammonium bromide, Triton X-100, and sodium deoxycholate, which are usually used to split virus particles into separate structural proteins.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

David

Volokhov

et al. 2010

Appl Environ Microbiol

View full text Add to dashboard Cite

Although mycoplasmas are generally considered to be harmless commensals, some mycoplasma species are able to cause infections in pediatric, geriatric, or immunocompromised patients. Thus, accidental contamination of biologics with mycoplasmas represents a potential risk for the health of individuals who receive cell-derived biological and pharmaceutical products. To assess the efficiency of inactivation of mycoplasmas by the agents used in the manufacture of egg-derived influenza vaccines, we carried out a series of experiments aimed at monitoring the viability of mycoplasmas spiked into both chicken allantoic fluid and protein-rich microbiological media and then treated with beta-propiolactone, formalin, cetyltrimethylammonium bromide, Triton X-100, and sodium deoxycholate, which are agents that are commonly used for virus inactivation and disruption of viral particles during influenza vaccine production. Twenty-two mycoplasma species (with one to four strains of each species) were exposed to these inactivating agents at different concentrations. The most efficient inactivation of the mycoplasmas evaluated was observed with either 0.5% Triton X-100 or 0.5% sodium deoxycholate. Cetyltrimethylammonium bromide at concentrations of >0.08% was also able to rapidly inactivate (in less than 30 min) all mycoplasmas tested. In contrast, negligible reductions in mycoplasma titers were observed with 0.0125 to 0.025% formaldehyde. However, increasing the concentration of formaldehyde to 0.1 to 0.2% improved the mycoplasmacidal effect. Incubation of mycoplasmas with 0.1% beta-propiolactone for 1 to 24 h had a marked mycoplasmacidal effect. A comparison of the mycoplasma inactivation profiles showed that strains of selected species (Mycoplasma synoviae, Mycoplasma gallisepticum, Mycoplasma orale, Mycoplasma pneumoniae, and Acholeplasma laidlawii) represent a set of strains that can be utilized to validate the effectiveness of mycoplasma clearance obtained by inactivation and viral purification processes used for the manufacture of an inactivated egg-based vaccine.The mycoplasmas (the trivial name for Mollicutes) are a class of broadly distributed wall-less bacteria with some of the smallest known genomes that support bacterial self-replication (43). While most mycoplasmas are naturally harmless commensals that colonize a wide range of plant, insect, reptilian, avian, mammalian, and human hosts (43), some of them are able to cause diseases whose severity varies in their natural hosts (1,4,10,19,43).Mycoplasmas, particularly species of genera Mycoplasma and Acholeplasma, are also frequent contaminants of primary and continuous cell lines (45). This is a concern for research laboratories and commercial facilities involved in the development and production of cell-derived biological and pharmaceutical products. In addition, several natural mycoplasma infections of embryonated fowl eggs and primary chicken embryo cell cultures used for the production of viral vaccines have been reported (5-7, 16, 36, 46, 48, 55). Although only a ...

show abstract

Section: Discussionsupporting

confidence: 68%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

David

Volokhov

et al. 2010

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…This is particularly relevant at present since inactivated polio vaccines are going to be the vaccines of choice for the endgame of polio eradication, and changes in the manufacturing process, such as the use of different poliovirus strains as vaccine seeds, are expected. Manufacturers are also considering the use of alternative chemicals such as beta-propiolactone and binary ethyleneimine for inactivation since they are used for the production of other viral vaccines such as influenza virus, rabies virus, and foot-and-mouth disease virus (62)(63)(64). Analysis of the protein and RNA viral properties, as shown here, could help in assessing the impact of changes in antigenicity and stability of candidate vaccines, as well as in detecting residual infectivity during the inactivation process.…”

Section: Discussionmentioning

confidence: 99%

Effect of Formaldehyde Inactivation on Poliovirus

Wilton

Dunn

Eastwood

et al. 2014

J Virol

View full text Add to dashboard Cite

Inactivated polio vaccines, which have been used in many countries for more than 50 years, are produced by treating live poliovirus (PV) with formaldehyde. However, the molecular mechanisms underlying virus inactivation are not well understood. Infection by PV is initiated by virus binding to specific cell receptors, which results in viral particles undergoing sequential conformational changes that generate altered structural forms (135S and 80S particles) and leads to virus cell entry. We have analyzed the ability of inactivated PV to bind to the human poliovirus receptor (hPVR) using various techniques such as ultracentrifugation, fluorescence-activated cell sorting flow cytometry and real-time reverse transcription-PCR (RT-PCR). The results showed that although retaining the ability to bind to hPVR, inactivated PV bound less efficiently in comparison to live PV. We also found that inactivated PV showed resistance to structural conversion in vitro, as judged by measuring changes in antigenicity, the ability to bind to hPVR, and viral RNA release at high temperature. Furthermore, viral RNA from inactivated PV was shown to be modified, since cDNA yields obtained by RT-PCR amplification were severely reduced and no infectious virus was recovered after RNA transfection into susceptible cells. IMPORTANCEThis study represents a novel insight into the molecular mechanisms responsible for poliovirus inactivation. We show that inactivation with formaldehyde has an effect on early steps of viral replication as it reduces the ability of PV to bind to hPVR, decreases the sensitivity of PV to convert to 135S particles, and abolishes the infectivity of its viral RNA. These changes are likely responsible for the loss of infectivity shown by PV following inactivation. Techniques used in this study represent new approaches for the characterization of inactivated PV products and could be useful in developing improved methods for the production and quality control testing of inactivated polio vaccines. Measuring the antigenicity, capsid stability, and RNA integrity of inactivated PV samples could help establishing the optimal balance between the loss of infectivity and the preservation of virus antigenicity during inactivation.

show abstract

“…Virus used for immunisations was grown in HEp-2 cells (ATCC, Manassas, USA), harvested as clarified cell lysate in serum-free DMEM (Life Technologies, Paisley, UK) and inactivated by b-propiolactone treatment (Sigma) as described [36]. Mock virus preparations were obtained from uninfected HEp-2 cells.…”

Section: Immunisationsmentioning

confidence: 99%

Intranasal immunisation with inactivated RSV and bacterial adjuvants induces mucosal protection and abrogates eosinophilia upon challenge

et al. 2006

View full text Add to dashboard Cite

We have previously shown that following intranasal exposure to influenza virus, specific plasma cells are generated in the nasal-associated lymphoid tissue (NALT) and maintained for the life of the animal. However, we also showed that following infection with respiratory syncytial virus (RSV), specific plasma cells are generated in the NALT but wane quickly and are not maintained even after challenge, even though RSVspecific serum antibody responses remain robust. Only infection with influenza virus generated sterilising immunity, implying a role for these long-lived plasma cells in protection. We show here that the RSV-specific IgA NALT plasma cell population and lung antibody levels can be substantially boosted, both at acute and memory time points, by intranasal immunisation with inactivated RSV (iRSV) in combination with bacterial outer membrane vesicles (OMV) compared to live RSV alone. Finally, challenge with live RSV showed that immunisation with iRSV and OMV protect against both virus replication in the lung and the eosinophil infiltrate generated by either live RSV or iRSV alone. These data show that immunisation with iRSV and OMV maintains a NALT RSV-specific plasma cell population and generates an efficient protective immune response following RSV infection.See accompanying commentary: http://dx

show abstract

Principles of selective inactivation of viral genome. VI. Inactivation of the infectivity of the influenza virus by the action of β-propiolactone

Cited by 58 publications

References 9 publications

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Effect of Formaldehyde Inactivation on Poliovirus

Intranasal immunisation with inactivated RSV and bacterial adjuvants induces mucosal protection and abrogates eosinophilia upon challenge

Contact Info

Product

Resources

About