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1.

Figure 1. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

HBV consensus core DNA construct (pMCore). (A) Phylogenetic analysis of HBcAg consensus sequence as compare to individual genotypes (A through E). (B) Schematic representation of plasmid map and sequence of pMCore.

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.
2.
Figure 5

Figure 5. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

In vivospecific killing. Two groups of mice immunized with either pVax (control) or pMCore received CFSE-labeled target cells (CFSElo pulsed with irrelevant peptide or CFSEhi pulsed with epitope-specific peptide) through the tail vain. CFSE-labeled cells were recovered and analysis by FACS was utilized to quantify percent killing.

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.
3.

Figure 4. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

DNA immunization with pMCore drives strong antigen-specific immunity in liver of Balb/c mice. (A) Percent HBcAg-specific CD4 and CD8 IFN-γ+, TNF-α+ producing cells in the liver. (B) Average percent HBcAg-specific CD4 or CD8 double positive producing cells. (C) Antigen-specific antibody producing splenocytes. The values are the means ± standard error of the mean. Significance was determined by Student’s t test.

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.
4.
Figure 2

Figure 2. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

Detection of pMCore expression via invitro translation and immunofluorescence. (A) Transcription/translation reaction using pMCore plasmid was immuno-precipitated with anti-HA monoclonal antibody which recognize a HA epitope encoded into the pMCore antigen. The precipitate was run on a SDS-PAGE gel. (B) pMCore was detected in transiently transfected cells using a primary monoclonal HA tag antibody followed by detection with DyLight 594-labeled anti-rabbit secondary antibody (red). Hoechst stain was also used to fluorescently label cell nuclei. Expression of MCore is mostly localized to the cytoplasm as shown by the green stating patterns concentration outside the nucleus

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.
5.

Figure 3. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

Immunization with pMCore induces strong HBcAg-specific immunity in splenocytes of Balb/c mice. (A) Immunization scheme. 4 mice were immunized intramuscularly with 30μg pMCore. (B) Frequency of HBcAg-specific IFN-γ spot forming units (SFU) per million splenocytes after stimulation of spleen cells from immunized mice. (C) Average percent HBcAg-specific CD4 or CD8 IFN-γ+, TNF-α+ (D) Average percent HBcAg-specific CD4 or CD8 double positive producing cells. (E) Percent proliferation of CD4 and CD8 T cells (F) HBcAg-specific humoral immune response induced by pMCore. The values are the means ± standard error of the mean. Significance was determined by Student’s t test.

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.
6.

Figure 6. From: Synthetic DNA Immunogen encoding Hepatitis B core antigen drives immune response in liver.

Degranulation and elimination of HBcAg-expressing hepatocytes by vaccine-primed T cells. Hepatocytes of pMCore-immunized mice were transiently transfected with HBcAg or HCV NS3/4A plasmid. (A) Immunostaining of liver sections taken three days after hydrodynamic injection of PBS, pMCore (HBcAg) or pNS3/4A (HCV NS3-4A) from naïve or mice that were immunized with pMcore is shown. Clearance is much higher for the pMCore-immunized liver as compared to the NS3/4a transfected control liver. pMcore or NS3/4A expression detected with an anti-HA antibody (green cells) (B) 3 days post transfection, Cells were analyzed for degranulation marker expression, CD107a, and IFN-γ+ expression following stimulation with HBcAg peptides. (C) Serum ALT levels at day 3, 6 and 12 post transfection were measured and show no evidence of elevation in relevant vaccinated animals.

Nyamekye Obeng-Adjei, et al. Cancer Gene Ther. ;19(11):779-787.

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