Display Settings:

Items per page

Results: 7

1.
FIG. 2.

FIG. 2. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Detection of siRNA-mediated gene knockdown in vitro, using a bioluminescence imaging system. BHK-21 cells were transfected with 0.01 μg of pcDNA3-Luc and DNA encoding luciferase-specific shRNA (pRS-Luc) in amounts of 0.00, 0.01, 0.03, 0.09, 0.27, and 0.81 μg at 24- and 48-hr time points. Cells transfected with pcDNA3-Luc and pRS vector alone (0.00 μg of pRS-Luc) were used as the control. (A) Luminescent images of transfected cells at 24 hr (left) and 48 hr (right) posttransfection. (B) Line graph depicting the luminescence intensities over time of cells transfected with 0.01 μg of pcDNA3-Luc and various amounts of pRS-Luc. Results are expressed as the mean luminescence intensity and SD. Note: Luciferase expression in shRNA-transfected cells was suppressed in a dose-dependent manner and was still substantially reduced after 120 hr.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
2.
FIG. 1.

FIG. 1. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Bioluminescence imaging of C57BL/6J mice administered DNA encoding the luciferase gene. C57BL/6J mice (two per group) were intradermally administered two bullets containing 1 μg of plasmid DNA, each encoding luciferase (pcDNA3-Luc) or control (pcDNA3), via gene gun and luciferase activity was monitored over a period of 5 days by bioluminescence imaging. (A) Representative luminescence images demonstrating in vivo luciferase expression 30 min after gene gun administration in luciferase DNA-administered mice (left) and control DNA-administered mice (right). (B) Bar graph of luminescence intensity over a period of 144 hr after gene gun administration. Results are expressed as means and SD. Note: Luciferase activity peaked 24 hr postadministration and then gradually declined.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
3.
FIG. 3.

FIG. 3. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Bioluminescence imaging of luciferase activity in vivo. C57BL/6J mice (five per group) were administered intradermally, via gene gun, 0.5 μg of pcDNA3-Luc plus empty pRS vector (left) or 0.5 μg of pcDNA3-Luc plus pRS-Luc (right) and luciferase activity was monitored periodically by bioluminescence imaging. (A) Representative images of luciferase expression in mice 24 hr after gene gun administration. (B) Bar graph depicting luminescence intensity in vaccinated mice 2, 24, and 48 hr after gene gun administration. Results are expressed as mean luminescence intensity and SD. p Values were calculated by Student t test (*p < 0.01). Note: Levels of luciferase expression were significantly lower in mice administered DNA encoding pcDNA3-Luc plus pRS-Luc compared with mice administered DNA encoding pcDNA3-Luc plus pRS-vector control.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
4.
FIG. 7.

FIG. 7. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Characterization of apoptotic cell death of E7-specific CD8 T cells after incubation with E7 peptide-loaded DC-1 cells expressing FasL. E7-specific CD8+ T cells (2 × 105) were incubated in a 24-well plate with or without 1 × 107 E7 peptide-loaded DC-1 cells transfected with FasL-GFP at a 1:50 ratio in the presence of FasL-blocking antibody or isotype control (without FasL blocking). The percentage of apoptotic cells was characterized with antibodies specific for activated caspase-3 and analyzed by flow cytometric analysis. Bar graph depicts the average percentage of activated caspase-3-positive T cell detected in wells containing the indicated cells and blocking antibodies (means and SD). p Values were calculated by Student t test. Data are representative of two separate analyses.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
5.
FIG. 4.

FIG. 4. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Flow cytometric analysis demonstrating expression of FasL in BHK-21 cells in vitro. BHK-21 cells were transfected with FasL DNA and DNA encoding either nonspecific shRNA or FasL-specific shRNA. Cells were then stained with FasL-specific antibody or an isotype control (data not shown) and subjected to flow cytometric analysis to characterize FasL expression. Cells that did not receive FasL transfection were measured as a negative control. (A) Flow cytometric data representative of three separate analyses. The number shown in each histogram provides an indication of the percentage of gated cells staining positive for FasL. (B) Bar graph depicting relative FasL expression. Data are shown as means and SD. p Values were calculated by Student t test (*p < 0.05). Note: Within the experimental groups transfected with FasL DNA, FasL expression is significantly reduced in cells treated with DNA encoding FasL-specific shRNA compared with cells treated with DNA encoding nonspecific shRNA.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
6.
FIG. 6.

FIG. 6. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

In vivo treatment experiment involving tumor-challenged mice vaccinated with a combination of Sig/E7/LAMP-1 DNA and DNA encoding shRNAs. C57BL/6J mice (five per group) were challenged subcutaneously with TC-1/Luciferase tumor cells (5 × 104 cells per mouse). Eight days after tumor challenge, mice were treated with mixtures of Sig/E7/LAMP-1 DNA vaccine combined with DNA encoding a nonspecific shRNA, or FasL shRNA, delivered by gene gun to the shaved abdomen, and then boosted with the same vaccine three times at 4-day intervals. Tumor growth was monitored regularly by luminescence imaging with the Xenogen IVIS system. Bioluminescence signals were acquired for 30 sec. (A) Luminescence imaging depicting tumor load in animals bearing subcutaneously injected TC-1/Luciferase tumor cells 7 days after tumor challenge (1 day before commencement of gene gun vaccination therapy), and 15 days (for untreated) or 31 days after tumor challenge. (B) Line graphs depicting growth progression of tumors in individual mice from each treatment group from day 2, until day 15 (no vaccine group), or day 31, as measured by quantification of tumor bioluminescence intensity.

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.
7.
FIG. 5.

FIG. 5. From: RNA Interference-Mediated In Vivo Silencing of Fas Ligand as a Strategy for the Enhancement of DNA Vaccine Potency.

Analysis of IFN-γ-secreting E7-specific CD8+ T cells in mice vaccinated with Sig/E7/LAMP-1 DNA and DNA encoding shRNA targeting FasL. C57BL/6J mice (three per group) were vaccinated intradermally via gene gun with Sig/E7/LAMP-1 DNA in combination with DNA encoding nonspecific shRNA or DNA encoding FasL shRNA. Splenocytes were obtained from vaccinated mice and cultured with E7 peptide (amino acids 49–57) overnight. The cells were then analyzed for CD8 and intracellular IFN-γ staining by flow cytometry. (A) Representative flow cytometric data showing the number of IFN-γ+CD8+ T cells in vaccinated mice (top right quadrant). Data shown are representative of two separate analyses. (B) Bar graph showing the average fold increase in IFN-γ+ CD8+ T cells from each group of vaccinated mice. p Values were calculated by Student t test (*p < 0.005). Note: Vaccination with DNA encoding FasL-specific shRNA increased the number of E7-specific IFN-γ-secreting CD8 T cells by more than 2-fold. (C) CTL assay using luciferase-expressing TC-1 cells as target cells. Splenocytes from the various vaccinated groups were stimulated with E7 peptide (amino acids 49–57) in vitro. Target cells were incubated with the E7 peptide-stimulated splenocytes. Media from the incubated cells were collected and characterized for luminescence activity. Bar graph depicts the luminescence activity of the medium (means and SD).

Bruce Huang, et al. Hum Gene Ther. 2008 August;19(8):763-773.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk