Expression of membrane-bound and secreted forms of equine herpesvirus 1 glycoprotein D by recombinant baculovirus

Flowers, C. Clay; Flowers, Scarlett P.; Sheng, Yiwei; Tarbet, E. Bart; Jennings, Stephen R.; O’Callaghan, Dennis J.

doi:10.1016/0168-1702(94)00075-n

Cited by 11 publications

(4 citation statements)

References 25 publications

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“…The EHV-1 gD was expressed by using a recombinant baculovirus and two bands were observed probably due to degradation products or different stages of glycosylation or translation of gD. These findings are in line with those found by other authors, who suggested that the presence of more than one band may be due to incomplete translation or proteolytic degradation (Flowers et al, 1995;Love et al, 1993). After purification, two bands were also detected.…”

supporting

confidence: 88%

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

Fuentealba

Sguazza

Scrochi

et al. 2014

Journal of Virological Methods

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supporting

confidence: 88%

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

Fuentealba

Sguazza

Scrochi

et al. 2014

Journal of Virological Methods

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“…EHV-1 gD, a 55-58-kDa glycoprotein, is a component of the virion envelope, is inserted into the membrane of EHV-1-infected cells, and is required for virus entry and cell-tocell spread (14,16,53). Glycoprotein D of alphaherpesviruses, including EHV-1, elicits virus-specific B-and T-cell responses that are often associated with protective immunity in vivo (5,27,40,42,49,50,52,56).…”

Section: Discussionmentioning

confidence: 99%

Cytokine Profiles and Long-Term Virus-Specific Antibodies Following Immunization of CBA Mice with Equine Herpesvirus 1 and Viral Glycoprotein D

Zhang

Frampton²,

Osterrieder³

et al. 2003

Viral Immunology

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Equine herpesvirus 1 (EHV-1)-specific antibody-secreting cells (ASC) isolated from the lung and spleen of mice at 12 months after immunization with attenuated EHV-1 KyA, heat-killed KyA, or recombinant viral glycoprotein D (rgD) assessed by ELISPOT showed a three- to fivefold increase in three immunoglobulin isotypes at 3 days post-challenge with pathogenic EHV-1 RacL11 as compared to control mice. ELISPOT assays demonstrated a high frequency of cells secreting proinflammatory tumor necrosis factor-alpha (TNF-alpha), interferon gamma (IFN-gamma), and interleukin 4 (IL-4) in the lungs in response to infection with KyA or RacL11 or immunization with rgD. Cytokine production elicited by EHV-1 KyA or RacL11 infection revealed similar frequencies of EHV-1-specific IFN-gamma and IL-4 spot forming cells in the mediastinal lymph nodes and spleen. However, KyA induced significantly greater amounts of IFN-gamma producing cells in the lungs than did RacL11. Intranasal immunization with KyA or rgD induced long-term immunity that provided protection against pathogenic EHV-1 challenge infection at 12 months post-immunization. Overall, the data indicate that immunization with infectious KyA or rgD induces significant levels of cytokines, virus-specific ASC in the lungs and spleen, and long-term virus specific B-cell responses.

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“…Preparation of EHV-1-encoding plasmids. The EHV-1 glycoproteins (gB, gC, gD, gE, gH, gI, gL), the IE protein and the UL5 early protein have been characterized extensively (Allen & Yeargan, 1987;Audonnet et al, 1990;Birch-Machin et al, 2000;Crabb et al, 1991;Elton et al, 1991;Flowers et al, 1995;Koen et al, 2000;Kukreja et al, 1998;Osterrieder et al, 1995;Stokes et al, 1996;Tewari et al, 1994;Wellington et al, 1996a, b;Whalley et al, 1995). The genes encoding the gC, gD, gE, gH, gI and gL glycoproteins, the IE protein and the UL5 protein were cloned into the PCR3.1 plasmid (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…It has been proposed that EHV-1 proteins expressed early in the replication cycle, such as those encoded by the IE and early (UL5) genes, are likely to be important CTL targets (Koen et al, 2000;Ruitenberg et al, 1999b;Siedek et al, 1999;Smith et al, 1998). In addition, the glycoproteins of herpesviruses have been identified as containing CTL epitopes (Gallichan et al, 1993), and several studies in mice have reported protection from EHV-1 infection by vaccination with gB, gC, gD, gH and gL glycoproteins (Allen & Yeargan, 1987;Crabb et al, 1991;Flowers et al, 1995;Kukreja et al, 1998;Osterrieder et al, 1995;Stokes et al, 1996;Wellington et al, 1996a, b;Whalley et al, 1995). The aim of our study was to identify EHV-1 genes that encode proteins containing CTL epitopes recognized by horses and to determine the ELA-A restriction of these CTL responses.…”

Section: Introductionmentioning

confidence: 99%

Identification of equine herpesvirus-1 antigens recognized by cytotoxic T lymphocytes

Soboll

Whalley

Koen

et al. 2003

Journal of General Virology

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Equine herpesvirus-1 (EHV-1) causes serious disease in horses throughout the world, despite the frequent use of vaccines. CTLs are thought to be critical for protection from primary and reactivating latent EHV-1 infections. However, the antigen-specificity of EHV-1-specific CTLs is unknown. The aim of this study was to identify EHV-1 genes that encode proteins containing CTL epitopes and to determine their MHC I (or ELA-A in the horse) restriction. Equine dendritic cells, transfected with a series of EHV-1 genes, were used to stimulate autologous CTL precursor populations derived from previously infected horses. Cytotoxicity was subsequently measured against EHV-1-infected PWM lymphoblast targets. Dendritic cells were infected with EHV-1 (positive control) or transfected with plasmids encoding the gB, gC, gD, gE, gH, gI, gL, immediateearly (IE) or early protein of EHV-1 using the PowderJect XR-1 research device. Dendritic cells transfected with the IE gene induced CTL responses in four of six ponies. All four of these ponies shared a common ELA-A3.1 haplotype. Dendritic cells transfected with gC, gD, gI and gL glycoproteins induced CTLs in individual ponies. The cytotoxic activity was ELA-A-restricted, as heterologous targets from ELA-A mismatched ponies were not killed and an MHC I blocking antibody reduced EHV-1-specific killing. This is the first identification of an EHV-1 protein containing ELA-A-restricted CTL epitopes. This assay can now be used to study CTL specificity for EHV-1 proteins in horses with a broad range of ELA-A haplotypes, with the goal of developing a multi-epitope EHV-1 vaccine.

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Expression of membrane-bound and secreted forms of equine herpesvirus 1 glycoprotein D by recombinant baculovirus

Cited by 11 publications

References 25 publications

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

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

Cytokine Profiles and Long-Term Virus-Specific Antibodies Following Immunization of CBA Mice with Equine Herpesvirus 1 and Viral Glycoprotein D

Identification of equine herpesvirus-1 antigens recognized by cytotoxic T lymphocytes

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