Evaluation by Polymerase Chain Reaction on the Effect of Betapropiolactone and Binary Ethyleneimine on DNA

Groseil, Catherine; Guérin, Philippe; Adamowicz, P

doi:10.1006/biol.1995.0035

Cited by 8 publications

(5 citation statements)

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“…Binary ethylenimine is an aziridine preparation commonly used in veterinary vaccine production as an inactivating agent (73). At 1 mM, BEI induces an alkylation of the nucleic acids without damaging proteins (74). In the present work, high ROS production coupled with upregulation of the five tested genes were recorded when using OsHV-1 inactivated with BEI at [0.04%] for 22h, supporting the role of viral proteins in the stimulation of antiviral response in oysters.…”

Section: Discussionsupporting

confidence: 77%

Inactivated ostreid herpesvirus-1 induces an innate immune response in the Pacific oyster, Crassostrea gigas, hemocytes

2023

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Infectious diseases are a major constraint to the expansion of shellfish production worldwide. Pacific oyster mortality syndrome (POMS), a polymicrobial disease triggered by the Ostreid herpesvirus-1 (OsHV-1), has devastated the global Pacific oyster (Crassostrea gigas) aquaculture industry. Recent ground-breaking research revealed that C. gigas possess an immune memory, capable of adaption, which improves the immune response upon a second exposure to a pathogen. This paradigm shift opens the door for developing ‘vaccines’ to improve shellfish survival during disease outbreaks. In the present study, we developed an in-vitro assay using hemocytes – the main effectors of the C. gigas immune system – collected from juvenile oysters susceptible to OsHV-1. The potency of multiple antigen preparations (e.g., chemically and physically inactivated OsHV-1, viral DNA, and protein extracts) to stimulate an immune response in hemocytes was evaluated using flow cytometry and droplet digital PCR to measure immune-related subcellular functions and gene expression, respectively. The immune response to the different antigens was benchmarked against that of hemocytes treated with Poly (I:C). We identified 10 antigen preparations capable of inducing immune stimulation in hemocytes (ROS production and positively expressed immune- related genes) after 1 h of exposure, without causing cytotoxicity. These findings are significant, as they evidence the potential for priming the innate immunity of oysters using viral antigens, which may enable cost-effective therapeutic treatment to mitigate OsHV-1/POMS. Further testing of these antigen preparations using an in-vivo infection model is essential to validate promising candidate pseudo-vaccines.

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Section: Discussionsupporting

confidence: 77%

Inactivated ostreid herpesvirus-1 induces an innate immune response in the Pacific oyster, Crassostrea gigas, hemocytes

2023

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“…Next, we wished to compare ORFV preparations which had been inactivated by binary ethyleneimine (BEI) or by heat inactivation. BEI is known to alkylate nucleic acids, thereby significantly damaging DNA, but reacts very little with proteins (Groseil et al , 1995). BEI and heat-inactivated ORFV preparations did not differ in their antiviral activity ().…”

Section: Resultsmentioning

confidence: 99%

Characterization of immunostimulatory components of orf virus (parapoxvirus ovis)

Friebe

Friederichs

Scholz

et al. 2011

Journal of General Virology

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Inactivated orf virus (ORFV, parapoxvirus ovis) induces antiviral activity in animal models of acute and chronic viral infections and exerts strong effects on human immune cells. ORFV activates antigen presenting cells (APC) via CD14 and, probably, Toll-like receptor signalling, and triggers the release of IFN-c that has been identified as the key mediator of the antiviral activity. After delineating virus proteins as being the most likely active constituent, we aimed to characterize the ORFV proteins responsible for the therapeutic effect. By using a vaccinia virus/ORFV expression library we identified several multi-gene DNA fragments with strong immunomodulatory activity. Together these fragments contain 27 ORFs. The encoded proteins are related to virion structure and transcription but are otherwise unrelated. Two proteins were separately expressed and purified, and demonstrated immunostimulatory activity. Gene expression profiles induced by ORFV and the identified fragments were investigated by microarray analysis. Interestingly, all active fragments induced a similar gene-expression pattern, differing only in quantitative aspects. Obviously, several proteins of ORFV activate similar cellular pathways, modulating APC to generate a strong T-helper 1-dominated immune response. This was balanced by additional induction of immune dampening mechanisms, suggesting regulatory differences compared to single cytokine therapies. We conclude that ORFV may have the potential to enrich the armamentarium of antiviral therapies.

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“…Binary ethylenimine (BEI) is a kind of aziridines and formed by the cyclization of 2-chloroethylamine hydrochloride (BEA). BEI is known to alkylate nucleic acids but do not damage the protein of inactivated virus in the concentration of 1 mM [28,29]. In veterinary medicine BEI is the preferred inactivating agent for producing vaccines containing animal viruses with DNA or RNA genomes [30][31][32].…”

Section: Introductionmentioning

confidence: 99%

Protection of Procambarus clarkii against white spot syndrome virus using inactivated WSSV

Zhu

Miao

et al. 2009

Fish & Shellfish Immunology

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Evaluation by Polymerase Chain Reaction on the Effect of Betapropiolactone and Binary Ethyleneimine on DNA

Cited by 8 publications

References 0 publications

Inactivated ostreid herpesvirus-1 induces an innate immune response in the Pacific oyster, Crassostrea gigas, hemocytes

Inactivated ostreid herpesvirus-1 induces an innate immune response in the Pacific oyster, Crassostrea gigas, hemocytes

Characterization of immunostimulatory components of orf virus (parapoxvirus ovis)

Protection of Procambarus clarkii against white spot syndrome virus using inactivated WSSV

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