Comparative studies of HSV‐1 antigens solubilised from infected cells by using non‐ionic or zwitterionic detergents

Jennings, R.; Ertürk, Murat

doi:10.1002/jmv.1890310206

Cited by 14 publications

(4 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The final glycoprotein preparation was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with rabbit anti-HSV-1-horseradish peroxidase (HRP) conjugate and detection by chemiluminescence using the ECL kit (Amersham). The presence of at least four HSV-1 glycoproteins, namely, gB, gC, gD, and gE, in such preparations was shown using glycoprotein-specific monoclonal antibodies in ELISA and by radioimmunoprecipitation (17). Mockinfected preparations were made from cell cultures in the same way described above but without the addition of virus.…”

Section: Methodsmentioning

confidence: 99%

Protective Mucosal Immunity to Ocular Herpes Simplex Virus Type 1 Infection in Mice by UsingEscherichia coliHeat-Labile Enterotoxin B Subunit as an Adjuvant

Richards

Aman

Hirst

et al. 2001

J Virol

View full text Add to dashboard Cite

The potential of nontoxic recombinant B subunits of cholera toxin (rCtxB) and its close relative Escherichia coli heat-labile enterotoxin (rEtxB) to act as mucosal adjuvants for intranasal immunization with herpes simplex virus type 1 (HSV-1) glycoproteins was assessed. Doses of 10 g of rEtxB or above with 10 g of HSV-1 glycoproteins elicited high serum and mucosal anti-HSV-1 titers comparable with that obtained using CtxB (10 g) with a trace (0.5 g) of whole toxin (Ctx-CtxB). By contrast, doses of rCtxB up to 100 g elicited only meager anti-HSV-1 responses. As for Ctx-CtxB, rEtxB resulted in a Th2-biased immune response with high immunoglobulin G1 (IgG1)/IgG2a antibody ratios and production of interleukin 4 (IL-4) and IL-10 as well as gamma interferon by proliferating T cells. The protective efficacy of the immune response induced using rEtxB as an adjuvant was assessed following ocular challenge of immunized and mock-immunized mice. Epithelial disease was observed in both groups, but the immunized mice recovered by day 6 whereas mock-immunized mice developed more severe corneal disease leading to stromal keratitis. In addition, a significant reduction in the incidence of lid disease and zosteriform spread was observed in immunized animals and there was no encephalitis compared with 95% encephalitis in mock-immunized mice. The potential of such mucosal adjuvants for use in human vaccines against pathogens such as HSV-1 is discussed.

show abstract

Section: Methodsmentioning

confidence: 99%

Protective Mucosal Immunity to Ocular Herpes Simplex Virus Type 1 Infection in Mice by UsingEscherichia coliHeat-Labile Enterotoxin B Subunit as an Adjuvant

Richards

Aman

Hirst

et al. 2001

J Virol

View full text Add to dashboard Cite

show abstract

“…This has been detailed elsewhere [11,22]. Briefly, HSV-1 infected Vero cells were disrupted using the zwitterionic detergent Empigen BB [2,23], and the lysate subjected to sonication, high-speed centrifugation and dialysis before gradient centrifugation.…”

Section: Preparation and Characterisation Of Hsv-1 Surface Antigensmentioning

confidence: 99%

Acute and latent infection of mice immunised with HSV-1 ISCOM vaccine

Ertürk

Hill

Shimeld³

et al. 1992

Archives of Virology

Self Cite

View full text Add to dashboard Cite

The effect of immunisation with an HSV-1 antigen preparation (containing at least 6 viral glycoproteins) on primary infection with HSV and the establishment of latency, was assessed in two mouse models (involving either skin or corneal challenge with virus). The vaccine preparation, given either with Freund's complete adjuvant or aluminium hydroxide gel or in the form of immunostimulating complexes (ISCOMS), induced high ELISA antibody responses (highest with HSV as the ISCOM preparation) and low levels of neutralising antibody. In both models, immunisation with the HSV ISCOM preparation significantly reduced the incidence of zosteriform spread of virus and the severity of disease and, in some cases, the incidence of latent infection in sensory ganglia. In the eye model it was possible to show that immunisation with the HSV ISCOMS restricted the establishment of latency almost entirely to the ophthalmic part of the trigeminal ganglion. Protection from establishment of latency correlated with prechallenge antibody levels.

show abstract

“…More importantly, it has been shown safe to prepare protein antigens for vaccine application. For example, Jennings and Eturk demonstrated in mice studies that Empigen-solubilized HSV-1 antigen preparations elicited greater antibody responses and greater protection against HSV-1 challenge than the NP40-solubilized preparation [40]. Lowthert et al found Empigen BB to be the most efficient detergent for the solubilization of keratins while preserving their antigenicity [41].…”

Section: Discussionmentioning

confidence: 99%

Purification of recombinant vaccinia virus-expressed monomeric HIV-1 gp120 to apparent homogeneity

Guo¹,

Cleveland²,

Davenport³

et al. 2013

Protein Expression and Purification

View full text Add to dashboard Cite

Vaccinia virus (VV) has been used to express a variety of heterologous proteins, including HIV envelope (Env) glycoproteins. The Env protein is synthesized as a precursor molecule, gp160, which is cleaved into the surface antigen gp120 and the transmembrane protein gp41. Even though production of gp160 by the VV expression system has been described, its use for gp120 production is not well documented. Here we report a new procedure for the purification of gp120 from serum-containing culture supernatant of VV infected cells. The gp120 protein was enriched to a purity better than 60% on a snowdrop (Galanthus nivalis) lectin affinity column in the presence of 0.25% zwitterionic detergent Empigen BB. After additional DEAE anion exchange and Superdex size exclusion chromatography steps, the gp120 monomer was purified free of contamination as determined by SDS-PAGE. The retention of structural integrity was confirmed by determining the affinity constant of purified gp120s to soluble CD4 and a monoclonal antibody IgG1b12, using surface plasmon resonance analysis. The purification procedure is robust and reproducible, and may find general use for glycoprotein purifications from sources where the presence of serum is desirable.

show abstract

Comparative studies of HSV‐1 antigens solubilised from infected cells by using non‐ionic or zwitterionic detergents

Cited by 14 publications

References 35 publications

Protective Mucosal Immunity to Ocular Herpes Simplex Virus Type 1 Infection in Mice by UsingEscherichia coliHeat-Labile Enterotoxin B Subunit as an Adjuvant

Protective Mucosal Immunity to Ocular Herpes Simplex Virus Type 1 Infection in Mice by UsingEscherichia coliHeat-Labile Enterotoxin B Subunit as an Adjuvant

Acute and latent infection of mice immunised with HSV-1 ISCOM vaccine

Purification of recombinant vaccinia virus-expressed monomeric HIV-1 gp120 to apparent homogeneity

Contact Info

Product

Resources

About