Thermal inactivation of adenovirus type 5

Maheshwari, Gargi; Jannat, Risat; McCormick, Lisa; Hsu, David S.

doi:10.1016/j.jviromet.2004.02.003

Cited by 35 publications

(22 citation statements)

References 9 publications

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“…This result is similar to the results in Fig. 3 showing inactivation of virus infectivity at 45°C, as well as previous data demonstrating virus inactivation at temperatures between 48 and 54°C [36].…”

Section: Resultssupporting

confidence: 93%

Genetic Incorporation of Human Metallothionein into the Adenovirus Protein IX for Non-Invasive SPECT Imaging

et al. 2011

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As the limits of existing treatments for cancer are recognized, clearly novel therapies must be considered for successful treatment; cancer therapy using adenovirus vectors is a promising strategy. However tracking the biodistribution of adenovirus vectors in vivo is limited to invasive procedures such as biopsies, which are error prone, non-quantitative, and do not give a full representation of the pharmacokinetics involved. Current non-invasive imaging strategies using reporter gene expression have been applied to analyze adenoviral vectors. The major drawback to approaches that tag viruses with reporter genes is that these systems require initial viral infection and subsequent cellular expression of a reporter gene to allow non-invasive imaging. As an alternative to conventional vector detection techniques, we developed a specific genetic labeling system whereby an adenoviral vector incorporates a fusion between capsid protein IX and human metallothionein. Our study herein clearly demonstrates our ability to rescue viable adenoviral particles that display functional metallothionein (MT) as a component of their capsid surface. We demonstrate the feasibility of 99mTc binding in vitro to the pIX-MT fusion on the capsid of adenovirus virions using a simple transchelation reaction. SPECT imaging of a mouse after administration of a 99mTc-radiolabeled virus showed clear localization of radioactivity to the liver. This result strongly supports imaging using pIX-MT, visualizing the normal biodistribution of Ad primarily to the liver upon injection into mice. The ability we have developed to view real-time biodistribution in their physiological milieu represents a significant tool to study adenovirus biology in vivo.

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Section: Resultssupporting

confidence: 93%

Genetic Incorporation of Human Metallothionein into the Adenovirus Protein IX for Non-Invasive SPECT Imaging

et al. 2011

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“…Also SARS-CoV could be inactivated after incubation of 1 h at 37°C at a pH2 or pH12 [14]. Incubation of PRRSV for 48 h at 37°C was efficient for its inactivation, while SARSCoV was inactivated after incubation of 20 min at 56°C [14] and adenovirus type 5 after 10 min at 50°C [33]. For this study, inactivation of PRRSV at higher temperatures was not considered, since PRRSV proteins, similar to what is observed with other viruses [56], will most likely be denaturated, thereby destroying important epitopes.…”

Section: Discussionmentioning

confidence: 96%

“…After incubation, the pH was neutralized [14]. For temperature inactivation, virus was incubated for different times at 37°C [14,33]. Inactivation of PRRSV with gamma irradiation was performed using an electron accelerator (Prof. L. Van Hoorebeke, Ghent University, Faculty of Science, Department of Subatomic and Radiation Physics).…”

Section: Inactivation Methodsmentioning

confidence: 99%

Assessing the functionality of viral entry-associated domains of porcine reproductive and respiratory syndrome virus during inactivation procedures, a potential tool to optimize inactivated vaccines

2009

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-Porcine reproductive and respiratory syndrome virus (PRRSV) causes severe economic losses in the pig industry worldwide. Currently, vaccines based on inactivated PRRSV provide limited protection of pigs against infection, most likely because viral epitopes associated with the induction of neutralizing antibodies are not or poorly conserved during inactivation. To analyze the effect of inactivation procedures on the interaction of PRRSV with receptors involved in virus entry, a new assay was set up in this study. Viral entry-associated domains are most likely important for the induction of neutralizing antibodies, since neutralizing antibodies block interaction of PRRSV with cellular receptors. To investigate the interaction of PRRSV with the cellular receptors upon different inactivation procedures, attachment to and internalization of inactivated PRRSV into macrophages were monitored. AT-2 could not inactivate PRRSV completely and is therefore not useful for vaccine development. PRRSV inactivated with ultraviolet light, binary ethyleneimine and gamma irradiation, which all mainly have an effect at the genomic level, showed no difference compared to control live virus at all levels of virus entry, whereas PRRSV treated with formaldehyde, glutaraldehyde and pH changes, which all have a modifying effect on proteins, was not able to internalize into macrophages anymore. These results suggest that inactivation with methods with a main effect on the viral genome preserve PRRSV entry-associated domains and are useful for future development of an effective inactivated vaccine against PRRSV. Although PRRSV incubation at 37°C can completely inactivate PRRSV with preservation of entry-associated domains, this method is not recommended for vaccine development, since the mechanism is yet unknown.PRRSV / inactivation / vaccine / entry-associated domain / sialoadhesin

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“…To test whether aggregated, uninfectious viral proteins labeled by the fluorophore were readily internalized by cells, we heated an aliquot of the fluorescently labeled virus preparation to 70°C for 30 min, conditions sufficient to render the Ad uninfectious (Maheshwari et al, 2004). We then incubated the heat-killed virus with HFF cells for 24 h, fixed the cells in methanol, and analyzed the cells for the presence of internalized AF-555 fluorescence.…”

Section: Conjugation Of Fluorescent Dye To Ad Particlesmentioning

confidence: 99%

Infection with Replication-deficient Adenovirus Induces Changes in the Dynamic Instability of Host Cell Microtubules

Warren

Rutkowski

Cassimeris

2006

MBoC

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Thermal inactivation of adenovirus type 5

Cited by 35 publications

References 9 publications

Genetic Incorporation of Human Metallothionein into the Adenovirus Protein IX for Non-Invasive SPECT Imaging

Genetic Incorporation of Human Metallothionein into the Adenovirus Protein IX for Non-Invasive SPECT Imaging

Assessing the functionality of viral entry-associated domains of porcine reproductive and respiratory syndrome virus during inactivation procedures, a potential tool to optimize inactivated vaccines

Infection with Replication-deficient Adenovirus Induces Changes in the Dynamic Instability of Host Cell Microtubules

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