Limits...
Co-Administration of Molecular Adjuvants Expressing NF-Kappa B Subunit p65/RelA or Type-1 Transactivator T-bet Enhance Antigen Specific DNA Vaccine-Induced Immunity.

Shedlock DJ, Tingey C, Mahadevan L, Hutnick N, Reuschel EL, Kudchodkar S, Flingai S, Yan J, Kim JJ, Ugen KE, Weiner DB, Muthumani K - Vaccines (Basel) (2014)

Bottom Line: Specifically the co-delivery of (a) RelA, a subunit of the NF-κB transcription complex or (b) T-bet, a Th1-specific T box transcription factor, along with a prototypical DNA vaccine expressing HIV-1 proteins was evaluated.As such, this study demonstrated that co-delivery of either adjuvant resulted in enhanced T and B cell responses, specifically characterized by increased T cell numbers, IFN-γ production, as well as enhanced antibody responses.This study demonstrates the use of cellular transcription factors as adjuvants for enhancing DNA vaccine-induced immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. shedlock@mail.med.upenn.edu.

ABSTRACT
DNA vaccine-induced immunity can be enhanced by the co-delivery of synthetic gene-encoding molecular adjuvants. Many of these adjuvants have included cytokines, chemokines or co-stimulatory molecules that have been demonstrated to enhance vaccine-induced immunity by increasing the magnitude or type of immune responses and/or protective efficacy. In this way, through the use of adjuvants, immune responses can be highly customizable and functionally tailored for optimal efficacy against pathogen specific (i.e., infectious agent) or non-pathogen (i.e., cancer) antigens. In the novel study presented here, we examined the use of cellular transcription factors as molecular adjuvants. Specifically the co-delivery of (a) RelA, a subunit of the NF-κB transcription complex or (b) T-bet, a Th1-specific T box transcription factor, along with a prototypical DNA vaccine expressing HIV-1 proteins was evaluated. As well, all of the vaccines and adjuvants were administered to mice using in vivo electroporation (EP), a technology demonstrated to dramatically increase plasmid DNA transfection and subsequent transgene expression with concomitant enhancement of vaccine induced immune responses. As such, this study demonstrated that co-delivery of either adjuvant resulted in enhanced T and B cell responses, specifically characterized by increased T cell numbers, IFN-γ production, as well as enhanced antibody responses. This study demonstrates the use of cellular transcription factors as adjuvants for enhancing DNA vaccine-induced immunity.

No MeSH data available.


Related in: MedlinePlus

Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env (A) or Gag (B) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated (3H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env (A, right panel) or Gag (B, right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. ****p < 0.0001.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4494262&req=5

vaccines-02-00196-f003: Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env (A) or Gag (B) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated (3H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env (A, right panel) or Gag (B, right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. ****p < 0.0001.

Mentions: Since the RelA molecular adjuvant was observed to particularly enhance T cell responses, the proliferative potential of cells immunized in the presence or absence of pRelA was evaluated. Splenocytes from vaccinated animals were harvested at seven days following the third immunization and were then stimulated with their cognate Ag, i.e., either HIV-1 Env or Gag (Figure 3). In pEnv-vaccinated mice, there was a trend towards enhanced proliferation at all Ag doses in mice that also received the pRelA adjuvant when compared to unadjuvanted animals (Figure 3A). This trend was also observed in pGag-vaccinated animals where the overall stimulation index was higher when pRelA was co-delivered (Figure 3B). As well, in both Figure 3A,B, in addition to the overall stimulation index, fold increase graphs are included, with the ‘fold” value being a ratio of stimulation index of the pEnv + pRelA or pGag + pRelA groups divided by stimulation indexes of the pEnv or pGag alone groups. Thus, the stimulation index in pEnv and pGag vaccinated animals was increased by the inclusion of a pRelA adjuvant, at all vaccine doses tested. These responses were specific for the HIV Ags since minimal proliferation was observed in splenocytes from animals that received the pRelA adjuvant alone. Taken together, these results demonstrate that the pRelA DNA adjuvant enhances Ag-specific T cell proliferative responses against two individual specific antigens.


Co-Administration of Molecular Adjuvants Expressing NF-Kappa B Subunit p65/RelA or Type-1 Transactivator T-bet Enhance Antigen Specific DNA Vaccine-Induced Immunity.

Shedlock DJ, Tingey C, Mahadevan L, Hutnick N, Reuschel EL, Kudchodkar S, Flingai S, Yan J, Kim JJ, Ugen KE, Weiner DB, Muthumani K - Vaccines (Basel) (2014)

Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env (A) or Gag (B) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated (3H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env (A, right panel) or Gag (B, right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. ****p < 0.0001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494262&req=5

vaccines-02-00196-f003: Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env (A) or Gag (B) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated (3H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env (A, right panel) or Gag (B, right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. ****p < 0.0001.
Mentions: Since the RelA molecular adjuvant was observed to particularly enhance T cell responses, the proliferative potential of cells immunized in the presence or absence of pRelA was evaluated. Splenocytes from vaccinated animals were harvested at seven days following the third immunization and were then stimulated with their cognate Ag, i.e., either HIV-1 Env or Gag (Figure 3). In pEnv-vaccinated mice, there was a trend towards enhanced proliferation at all Ag doses in mice that also received the pRelA adjuvant when compared to unadjuvanted animals (Figure 3A). This trend was also observed in pGag-vaccinated animals where the overall stimulation index was higher when pRelA was co-delivered (Figure 3B). As well, in both Figure 3A,B, in addition to the overall stimulation index, fold increase graphs are included, with the ‘fold” value being a ratio of stimulation index of the pEnv + pRelA or pGag + pRelA groups divided by stimulation indexes of the pEnv or pGag alone groups. Thus, the stimulation index in pEnv and pGag vaccinated animals was increased by the inclusion of a pRelA adjuvant, at all vaccine doses tested. These responses were specific for the HIV Ags since minimal proliferation was observed in splenocytes from animals that received the pRelA adjuvant alone. Taken together, these results demonstrate that the pRelA DNA adjuvant enhances Ag-specific T cell proliferative responses against two individual specific antigens.

Bottom Line: Specifically the co-delivery of (a) RelA, a subunit of the NF-κB transcription complex or (b) T-bet, a Th1-specific T box transcription factor, along with a prototypical DNA vaccine expressing HIV-1 proteins was evaluated.As such, this study demonstrated that co-delivery of either adjuvant resulted in enhanced T and B cell responses, specifically characterized by increased T cell numbers, IFN-γ production, as well as enhanced antibody responses.This study demonstrates the use of cellular transcription factors as adjuvants for enhancing DNA vaccine-induced immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. shedlock@mail.med.upenn.edu.

ABSTRACT
DNA vaccine-induced immunity can be enhanced by the co-delivery of synthetic gene-encoding molecular adjuvants. Many of these adjuvants have included cytokines, chemokines or co-stimulatory molecules that have been demonstrated to enhance vaccine-induced immunity by increasing the magnitude or type of immune responses and/or protective efficacy. In this way, through the use of adjuvants, immune responses can be highly customizable and functionally tailored for optimal efficacy against pathogen specific (i.e., infectious agent) or non-pathogen (i.e., cancer) antigens. In the novel study presented here, we examined the use of cellular transcription factors as molecular adjuvants. Specifically the co-delivery of (a) RelA, a subunit of the NF-κB transcription complex or (b) T-bet, a Th1-specific T box transcription factor, along with a prototypical DNA vaccine expressing HIV-1 proteins was evaluated. As well, all of the vaccines and adjuvants were administered to mice using in vivo electroporation (EP), a technology demonstrated to dramatically increase plasmid DNA transfection and subsequent transgene expression with concomitant enhancement of vaccine induced immune responses. As such, this study demonstrated that co-delivery of either adjuvant resulted in enhanced T and B cell responses, specifically characterized by increased T cell numbers, IFN-γ production, as well as enhanced antibody responses. This study demonstrates the use of cellular transcription factors as adjuvants for enhancing DNA vaccine-induced immunity.

No MeSH data available.


Related in: MedlinePlus