Production of equine herpesvirus 1 recombinant glycoprotein D and development of an agar gel immunodiffusion test for serological diagnosis

Fuentealba, Nadia Analía; Sguazza, Guillermo Hernán; Scrochi, Mariela Rita; Bravi, María Emilia; Zanuzzi, Carolina Natalia; Corva, Santiago Gerardo; Gimeno, Eduardo Juan; Pecoraro, Marcelo Ricardo Ítalo; Galosi, Cecília Mônica

doi:10.1016/j.jviromet.2014.02.025

Cited by 8 publications

(8 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…VC2-EHV-1-gD stimulated strong virus-neutralizing (VN) activity in comparison to that seen in both unvaccinated and VC2-vaccinated animals, and this VN activity was similar to that generated by the commercial EHV-1 killed virus vaccine. It has been reported that intranasal or intramuscular immunization with EHV-1 gD produces antigen-specific IgG responses (3,4,8,10,11,(22)(23)(24)(25). In addition, the presence of VN antibody prior to EHV-1 infection was shown to be associated with a reduction in the amount and duration of nasopharyngeal virus shedding (26,27).…”

Section: Figmentioning

confidence: 99%

“…EHV-1 glycoproteins facilitate multiple aspects of the viral life cycle, including mediation of the fusion of the viral envelope with cellular membranes, intracellular virion morphogenesis, egress, cell-to-cell spread, and virus-induced cell fusion. Thus, these proteins are major antigenic determinants for both humoral and cellular immune responses and have been utilized as subunit vaccines (2)(3)(4). Specifically, immunization with EHV-1 glycoprotein D (gD) has been shown to generate protective immune responses in mice and horses (3,(5)(6)(7)(8)(9)(10)(11).…”

mentioning

confidence: 99%

“…Thus, these proteins are major antigenic determinants for both humoral and cellular immune responses and have been utilized as subunit vaccines (2)(3)(4). Specifically, immunization with EHV-1 glycoprotein D (gD) has been shown to generate protective immune responses in mice and horses (3,(5)(6)(7)(8)(9)(10)(11). EHV-1 and herpes simplex virus 1 (HSV-1) glycoproteins D are required for entry into cells and cell-to-cell fusion, a function which is conserved in most but not all alphaherpesviruses (12).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Intramuscular Immunization of Mice with the Live-Attenuated Herpes Simplex Virus 1 Vaccine Strain VC2 Expressing Equine Herpesvirus 1 (EHV-1) Glycoprotein D Generates Anti-EHV-1 Immune Responses in Mice

Liu

Stanfield

Chouljenko

et al. 2017

J Virol

View full text Add to dashboard Cite

Vaccination remains the best option to combat equine herpesvirus 1 (EHV-1) infection, and several different strategies of vaccination have been investigated and developed over the past few decades. Herein, we report that the liveattenuated herpes simplex virus 1 (HSV-1) VC2 vaccine strain, which has been shown to be unable to enter into neurons and establish latency in mice, can be utilized as a vector for the heterologous expression of EHV-1 glycoprotein D (gD) and that the intramuscular immunization of mice results in strong antiviral humoral and cellular immune responses. The VC2-EHV-1-gD recombinant virus was constructed by inserting an EHV-1 gD expression cassette under the control of the cytomegalovirus immediate early promoter into the VC2 vector in place of the HSV-1 thymidine kinase (UL23) gene. The vaccines were introduced into mice through intramuscular injection. Vaccination with both the VC2-EHV-1-gD vaccine and the commercially available vaccine Vetera EHV XP 1/4 (Vetera; Boehringer Ingelheim Vetmedica) resulted in the production of neutralizing antibodies, the levels of which were significantly higher in comparison to those in VC2-and mock-vaccinated animals (P Ͻ 0.01 or P Ͻ 0.001). Analysis of EHV-1-reactive IgG subtypes demonstrated that vaccination with the VC2-EHV-1-gD vaccine stimulated robust IgG1 and IgG2a antibodies after three vaccinations (P Ͻ 0.001). Interestingly, Vetera-vaccinated mice produced significantly higher levels of IgM than mice in the other groups before and after challenge (P Ͻ 0.01 or P Ͻ 0.05). Vaccination with VC2-EHV-1-gD stimulated strong cellular immune responses, characterized by the upregulation of both interferonand tumor necrosis factor-positive CD4 ϩ T cells and CD8 ϩ T cells. Overall, the data suggest that the HSV-1 VC2 vaccine strain may be used as a viral vector for the vaccination of horses as well as, potentially, for the vaccination of other economically important animals.IMPORTANCE A novel virus-vectored VC2-EHV-1-gD vaccine was constructed using the live-attenuated HSV-1 VC2 vaccine strain. This vaccine stimulated strong humoral and cellular immune responses in mice, suggesting that it could protect horses against EHV-1 infection.KEYWORDS EHV-1, HSV-1, equine herpesvirus, live vector vaccines, glycoprotein D, immune response, mouse model

show abstract

Section: Figmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Intramuscular Immunization of Mice with the Live-Attenuated Herpes Simplex Virus 1 Vaccine Strain VC2 Expressing Equine Herpesvirus 1 (EHV-1) Glycoprotein D Generates Anti-EHV-1 Immune Responses in Mice

Liu

Stanfield

Chouljenko

et al. 2017

J Virol

View full text Add to dashboard Cite

show abstract

“…To overcome this issue, a type-specific ELISA targeting glycoprotein G (gG) was developed. 16,21 This ELISA, targeting at least 1 type-specific epitope in gG, can discriminate between the IgG immune response to EHV-1 and EHV-4. 13 Both type-common and type-specific ELISAs are commercially available.…”

Section: Virus Neutralization Testmentioning

confidence: 99%

A review of traditional and contemporary assays for direct and indirect detection of Equid herpesvirus 1 in clinical samples

2015

View full text Add to dashboard Cite

Equid herpesvirus 1 (EHV-1) is one of the most economically important equine viral pathogens. Its clinical manifestations in horses vary from acute upper respiratory tract disease, abortion, or neonatal death, to neurological disease termed equine herpesviral myeloencephalopathy, which may lead to paralysis and a fatal outcome. Successful identification of EHV-1 infection in horses depends on a variety of factors such as suitable case selection with emphasis on timing of sample collection, selection of appropriate sample(s) based on the clinical manifestations, application of relevant diagnostic technique(s) and/or test(s), and careful evaluation and interpretation of laboratory results. Several traditional serologic and virus isolation assays have been described; however, these assays have inherent limitations that prevent rapid and reliable detection of EHV-1. The advent of molecular biologic techniques has revolutionized the diagnosis of infectious diseases in humans and animal species. Specifically, polymerase chain reaction (PCR)-based assays have allowed detection of nucleic acid in clinical specimens precisely and rapidly as compared to the traditional methods that detect the agent or antigen, or agent-specific antibodies in serum. The new molecular methods, especially real-time PCR, can be a very useful means of EHV-1 detection and identification. Veterinarians involved in equine practice must be aware of the advantages and disadvantages of various real-time PCR assays, interpretation of viral genetic marker(s), and latency in order to provide the best standard of care for their equine patients.

show abstract

“…Glycoprotein D is also responsible for induction of neutralizing antibodies due to the presence of neutralizing epitopes (Maeda et al 1997). Also in other herpesvirus infections, gD was successfully used as antigen in diagnostic tests (Meskhat et al 2012;Fuentealba et al 2014).…”

Section: Discussionmentioning

confidence: 99%

The expression and serological reactivity of recombinant canine herpesvirus 1 glycoprotein D

2016

View full text Add to dashboard Cite

The aim of this work was to express recombinant glycoprotein D of canine herpesvirus 1 in bacterial cells and to evaluate its diagnostic sensitivity and specificity when compared to traditional serological methods. The gene fragment coding glycoprotein D of canine herpesvirus 1 was amplified by polymerase chain reaction, cloned into plasmid vector and expressed in Escherichia coli cells. Recombinant protein was then purified and used as an antigen in immunoblot for a detection of canine herpesvirus 1 specific antibodies. Antibody testing was performed on the panel of 100 canine sera by immunoblot with recombinant glycoprotein D as antigen and compared with indirect immunofluorescence assay. Serum samples were collected from 83 dogs with no history of canine herpesvirus 1 or reproductive disorders, and from 17 dogs from breeding kennels with a history of canine herpesvirus 1 related reproductive disorders. Sensitivity of glycoprotein D based immunoblot was 89.2% and specificity was 93%. Kappa value was calculated to be 0.8 between immunoblot and indirect immunofluorescence assay. Antibodies against canine herpesvirus 1 infection were detected in 33% of samples by immunoblot assay. Our study confirms that recombinant glycoprotein D expressed in bacterial cells could be used as a suitable and sensitive antigen for immunological tests and that herpesvirus infection seems to be common among the canine population in the Czech Republic.

show abstract

Production of equine herpesvirus 1 recombinant glycoprotein D and development of an agar gel immunodiffusion test for serological diagnosis

Cited by 8 publications

References 9 publications

Intramuscular Immunization of Mice with the Live-Attenuated Herpes Simplex Virus 1 Vaccine Strain VC2 Expressing Equine Herpesvirus 1 (EHV-1) Glycoprotein D Generates Anti-EHV-1 Immune Responses in Mice

Intramuscular Immunization of Mice with the Live-Attenuated Herpes Simplex Virus 1 Vaccine Strain VC2 Expressing Equine Herpesvirus 1 (EHV-1) Glycoprotein D Generates Anti-EHV-1 Immune Responses in Mice

A review of traditional and contemporary assays for direct and indirect detection of Equid herpesvirus 1 in clinical samples

The expression and serological reactivity of recombinant canine herpesvirus 1 glycoprotein D

Contact Info

Product

Resources

About