Journal:

Article Title: DNA Immunization against Herpes Simplex Virus: Enhanced Efficacy Using a Sindbis Virus-Based Vector

doi:

Figure Lengend Snippet: Schematic diagram of plasmid DNA-based expression vectors used for DNA immunization with HSV-1 gB (diagram is not drawn to scale). Individual elements comprising the functional expression cassettes for both conventional (pCI-gB) and Sindbis virus (pSIN1.5-gB and pSIN2.5-gB) plasmid vectors are indicated. Sindbis virus-derived sequences are shown in white and include the four nonstructural protein genes (nsPs), complete 5′- and 3′-end untranslated regions, subgenomic promoter (JR), and poly(A) tract (A40). Shaded regions depict the CMV immediate-early promoter (CMV) and intron (INT), hepatitis delta virus antigenomic ribozyme sequence (δ), simian virus 40 late-region transcription termination signal (TT) in the pCI vector, bovine growth hormone transcription termination signal (TT) in the pSIN vectors, hepatitis B virus posttranscriptional regulatory element (PRE), and HSV-1 gB.

Article Snippet: Western blotting was performed by reacting the membrane with a primary mouse monoclonal anti-HSV-1 gB antibody (ABi, Columbia, Md.), followed by a secondary goat anti-mouse IgG2a-horseradish peroxidase (HRP) antibody (Southern Biotechnology Associates, Birmingham, Ala.).

Techniques: Plasmid Preparation, Expressing, Functional Assay, Derivative Assay, Sequencing

Journal:

Article Title: DNA Immunization against Herpes Simplex Virus: Enhanced Efficacy Using a Sindbis Virus-Based Vector

doi:

Figure Lengend Snippet: Expression of HSV-1 gB in vitro from pCI and pSIN vectors. Cell lysates were prepared from BHK-21 cells transfected with the different plasmid vectors. Cell lysates were harvested 48 h posttransfection, separated on SDS–8 to 16% polyacrylamide gels, transferred, and probed with a mouse monoclonal anti-HSV-1 gB antibody followed by a goat anti-mouse IgG2a-HRP antibody. Sizes are indicated in kilodaltons.

Article Snippet: Western blotting was performed by reacting the membrane with a primary mouse monoclonal anti-HSV-1 gB antibody (ABi, Columbia, Md.), followed by a secondary goat anti-mouse IgG2a-horseradish peroxidase (HRP) antibody (Southern Biotechnology Associates, Birmingham, Ala.).

Techniques: Expressing, In Vitro, Transfection, Plasmid Preparation

Journal:

Article Title: DNA Immunization against Herpes Simplex Virus: Enhanced Efficacy Using a Sindbis Virus-Based Vector

doi:

Figure Lengend Snippet: Induction of HSV-1-specific total IgG antibody in mice immunized with pCI or pSIN vectors expressing gB. BALB/c mice given a single i.m. injection of the different vectors at either 3.0 or 0.3 μg/mouse were bled at day 14 postimmunization, immediately prior to lethal challenge with HSV-1 McKrae. ELISA was used to measure total IgG antibody titers in individual mice. The normal serum control represents a pool from several nonimmune mice.

Article Snippet: Western blotting was performed by reacting the membrane with a primary mouse monoclonal anti-HSV-1 gB antibody (ABi, Columbia, Md.), followed by a secondary goat anti-mouse IgG2a-horseradish peroxidase (HRP) antibody (Southern Biotechnology Associates, Birmingham, Ala.).

Techniques: Expressing, Injection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: DNA Immunization against Herpes Simplex Virus: Enhanced Efficacy Using a Sindbis Virus-Based Vector

doi:

Figure Lengend Snippet: Induction of HSV-1-specific total IgG antibody in mice immunized with pCI or pSIN vectors expressing gB. BALB/c mice given a single i.m. injection of the different vectors at 1.0, 0.3, or 0.05 μg/mouse were bled at 15 weeks postimmunization, and ELISA was used to measure total IgG antibody titers in individual mice. The normal serum control represents a pool from several nonimmune mice.

Article Snippet: Western blotting was performed by reacting the membrane with a primary mouse monoclonal anti-HSV-1 gB antibody (ABi, Columbia, Md.), followed by a secondary goat anti-mouse IgG2a-horseradish peroxidase (HRP) antibody (Southern Biotechnology Associates, Birmingham, Ala.).

Techniques: Expressing, Injection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: DNA Immunization against Herpes Simplex Virus: Enhanced Efficacy Using a Sindbis Virus-Based Vector

doi:

Figure Lengend Snippet: Induction of HSV-1 gB-specific CTL in mice immunized with pCI or pSIN vectors expressing gB. Splenocytes from individual mice collected after a single i.m. injection with the pCI-gB or pSIN-gB vectors were restimulated in vitro with BC-gB1 and tested for cytolytic activity in a 4-h 51Cr release assay using BC-gB1 target cells (closed symbols) and BC-βgal target cells (open symbols). Cytolytic activity is shown as the percent specific lysis detected at each effector/target cell ratio tested and represents the mean of triplicates. This experiment was repeated on two additional occasions with similar results.

Article Snippet: Western blotting was performed by reacting the membrane with a primary mouse monoclonal anti-HSV-1 gB antibody (ABi, Columbia, Md.), followed by a secondary goat anti-mouse IgG2a-horseradish peroxidase (HRP) antibody (Southern Biotechnology Associates, Birmingham, Ala.).

Techniques: Expressing, Injection, In Vitro, Activity Assay, Release Assay, Lysis