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Enhanced Delivery and Potency of Self-Amplifying mRNA Vaccines by Electroporation in Situ.

Cu Y, Broderick KE, Banerjee K, Hickman J, Otten G, Barnett S, Kichaev G, Sardesai NY, Ulmer JB, Geall A - Vaccines (Basel) (2013)

Bottom Line: Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications.In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein.These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

View Article: PubMed Central - PubMed

Affiliation: Novartis Vaccines & Diagnostics, Inc., 350 Massachusetts Ave, Cambridge, MA 02139, USA.

ABSTRACT
Nucleic acid-based vaccines such as viral vectors, plasmid DNA (pDNA), and mRNA are being developed as a means to address limitations of both live-attenuated and subunit vaccines. DNA vaccines have been shown to be potent in a wide variety of animal species and several products are now licensed for commercial veterinary but not human use. Electroporation delivery technologies have been shown to improve the generation of T and B cell responses from synthetic DNA vaccines in many animal species and now in humans. However, parallel RNA approaches have lagged due to potential issues of potency and production. Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications. In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein. These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

No MeSH data available.


Study timeline for assessment of self-amplifying mRNA vector in an HIV immunogenicity model. (A) Female Balb/C mice received 2 treatments of RNA or pDNA vectors with or without EP (RNA or pDNA, 1, 10 and 50 µg), MF59-adjuvanted gp140.SF162 HIV envelope protein (Env/MF59, 10 µg) or viral replicon particles (VRP, 1 × 107 IU) expressing the HIV Env-protein at weeks 0 and 3. All groups then received a single injection of 10 µg Env/MF59 at week 8. Serum and mucosal samples, and spleens for T-cell quantitation were collected on week 2 (2wp1), 5 (2wp2, pre-protein boost) 8 (5wp2) and 10 (2wp3, post-protein boost). Env-specific total IgG titer for all groups at 2wp1 and 2wp2 (B) and 5wp2 and 3wp2 (C) is plotted as log of mean titer, and the horizontal bar represents geometric mean titer (GMT). Titers < 25 (dotted line) were assigned a value of 5 for calculation of GMT. n = 6 at 2wp1, n = 12 at 2wp2, and n = 8 at 5wp2 and 2wp3. * denotes statistical significance with p < 0.05; ** denotes statistical significance with p < 0.005.
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vaccines-01-00367-f003: Study timeline for assessment of self-amplifying mRNA vector in an HIV immunogenicity model. (A) Female Balb/C mice received 2 treatments of RNA or pDNA vectors with or without EP (RNA or pDNA, 1, 10 and 50 µg), MF59-adjuvanted gp140.SF162 HIV envelope protein (Env/MF59, 10 µg) or viral replicon particles (VRP, 1 × 107 IU) expressing the HIV Env-protein at weeks 0 and 3. All groups then received a single injection of 10 µg Env/MF59 at week 8. Serum and mucosal samples, and spleens for T-cell quantitation were collected on week 2 (2wp1), 5 (2wp2, pre-protein boost) 8 (5wp2) and 10 (2wp3, post-protein boost). Env-specific total IgG titer for all groups at 2wp1 and 2wp2 (B) and 5wp2 and 3wp2 (C) is plotted as log of mean titer, and the horizontal bar represents geometric mean titer (GMT). Titers < 25 (dotted line) were assigned a value of 5 for calculation of GMT. n = 6 at 2wp1, n = 12 at 2wp2, and n = 8 at 5wp2 and 2wp3. * denotes statistical significance with p < 0.05; ** denotes statistical significance with p < 0.005.

Mentions: Self-amplifying mRNA vectors expressing the HIV envelope protein gp140.SF162 (Env) were used to vaccinate mice using a heterologous prime-boost regimen (Figure 3A). Mice were administered 2 doses of self-amplifying mRNA, as the prime, and a third vaccination with recombinant HIV Env protein adjuvanted with MF59 (Env/MF59), as the boost. Viral replicon particles (VRP) and pDNA expressing the same antigen were used as benchmarks. Serum and spleens were collected at the specified time points (Figure 3A). Antibody responses were measured by ELISA for total IgG (Figure 3B,C) and IgG isotypes (Figure 4), and T cell responses were evaluated using an intracellular cytokine immunofluorescence assay (Figure 5).


Enhanced Delivery and Potency of Self-Amplifying mRNA Vaccines by Electroporation in Situ.

Cu Y, Broderick KE, Banerjee K, Hickman J, Otten G, Barnett S, Kichaev G, Sardesai NY, Ulmer JB, Geall A - Vaccines (Basel) (2013)

Study timeline for assessment of self-amplifying mRNA vector in an HIV immunogenicity model. (A) Female Balb/C mice received 2 treatments of RNA or pDNA vectors with or without EP (RNA or pDNA, 1, 10 and 50 µg), MF59-adjuvanted gp140.SF162 HIV envelope protein (Env/MF59, 10 µg) or viral replicon particles (VRP, 1 × 107 IU) expressing the HIV Env-protein at weeks 0 and 3. All groups then received a single injection of 10 µg Env/MF59 at week 8. Serum and mucosal samples, and spleens for T-cell quantitation were collected on week 2 (2wp1), 5 (2wp2, pre-protein boost) 8 (5wp2) and 10 (2wp3, post-protein boost). Env-specific total IgG titer for all groups at 2wp1 and 2wp2 (B) and 5wp2 and 3wp2 (C) is plotted as log of mean titer, and the horizontal bar represents geometric mean titer (GMT). Titers < 25 (dotted line) were assigned a value of 5 for calculation of GMT. n = 6 at 2wp1, n = 12 at 2wp2, and n = 8 at 5wp2 and 2wp3. * denotes statistical significance with p < 0.05; ** denotes statistical significance with p < 0.005.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4494232&req=5

vaccines-01-00367-f003: Study timeline for assessment of self-amplifying mRNA vector in an HIV immunogenicity model. (A) Female Balb/C mice received 2 treatments of RNA or pDNA vectors with or without EP (RNA or pDNA, 1, 10 and 50 µg), MF59-adjuvanted gp140.SF162 HIV envelope protein (Env/MF59, 10 µg) or viral replicon particles (VRP, 1 × 107 IU) expressing the HIV Env-protein at weeks 0 and 3. All groups then received a single injection of 10 µg Env/MF59 at week 8. Serum and mucosal samples, and spleens for T-cell quantitation were collected on week 2 (2wp1), 5 (2wp2, pre-protein boost) 8 (5wp2) and 10 (2wp3, post-protein boost). Env-specific total IgG titer for all groups at 2wp1 and 2wp2 (B) and 5wp2 and 3wp2 (C) is plotted as log of mean titer, and the horizontal bar represents geometric mean titer (GMT). Titers < 25 (dotted line) were assigned a value of 5 for calculation of GMT. n = 6 at 2wp1, n = 12 at 2wp2, and n = 8 at 5wp2 and 2wp3. * denotes statistical significance with p < 0.05; ** denotes statistical significance with p < 0.005.
Mentions: Self-amplifying mRNA vectors expressing the HIV envelope protein gp140.SF162 (Env) were used to vaccinate mice using a heterologous prime-boost regimen (Figure 3A). Mice were administered 2 doses of self-amplifying mRNA, as the prime, and a third vaccination with recombinant HIV Env protein adjuvanted with MF59 (Env/MF59), as the boost. Viral replicon particles (VRP) and pDNA expressing the same antigen were used as benchmarks. Serum and spleens were collected at the specified time points (Figure 3A). Antibody responses were measured by ELISA for total IgG (Figure 3B,C) and IgG isotypes (Figure 4), and T cell responses were evaluated using an intracellular cytokine immunofluorescence assay (Figure 5).

Bottom Line: Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications.In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein.These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

View Article: PubMed Central - PubMed

Affiliation: Novartis Vaccines & Diagnostics, Inc., 350 Massachusetts Ave, Cambridge, MA 02139, USA.

ABSTRACT
Nucleic acid-based vaccines such as viral vectors, plasmid DNA (pDNA), and mRNA are being developed as a means to address limitations of both live-attenuated and subunit vaccines. DNA vaccines have been shown to be potent in a wide variety of animal species and several products are now licensed for commercial veterinary but not human use. Electroporation delivery technologies have been shown to improve the generation of T and B cell responses from synthetic DNA vaccines in many animal species and now in humans. However, parallel RNA approaches have lagged due to potential issues of potency and production. Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications. In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein. These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

No MeSH data available.