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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.


Histology of muscle tissues (transverse) by hematoxylin and eosin (H&E) staining, and expression of reporter protein (GFP) as delivered by intramuscular injection without EP (No EP) or with EP (+EP). Mice were injected intramuscularly with 50 µL of 1xPBS, pDNA or self-amplifying RNA vectors (pDNA or RNA, 5 µg/site). Images were obtained 2 days following treatment and are shown as representative of 6–8 slides taken per area, n = 4 muscles per group.
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vaccines-01-00367-f002: Histology of muscle tissues (transverse) by hematoxylin and eosin (H&E) staining, and expression of reporter protein (GFP) as delivered by intramuscular injection without EP (No EP) or with EP (+EP). Mice were injected intramuscularly with 50 µL of 1xPBS, pDNA or self-amplifying RNA vectors (pDNA or RNA, 5 µg/site). Images were obtained 2 days following treatment and are shown as representative of 6–8 slides taken per area, n = 4 muscles per group.

Mentions: To assess the effect of EP-enhanced delivery of nucleic acid vaccines in muscle, mice were injected intramuscularly with pDNA or self-amplifying mRNA (5 μg/site) with or without EP. The treated muscles were excised for GFP expression analysis and for hematoxylin and eosin (H&E) staining. Representative images of GFP expression in mouse muscle are shown in Figure 2. Robust GFP expression was detected in both pDNA + EP and RNA + EP treated groups. Low levels of GFP expression were detected in the RNA (no EP) control group, but not in the pDNA (no EP) or 1× PBS control groups. These results are consistent with that observed in the SEAP studies.


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)

Histology of muscle tissues (transverse) by hematoxylin and eosin (H&E) staining, and expression of reporter protein (GFP) as delivered by intramuscular injection without EP (No EP) or with EP (+EP). Mice were injected intramuscularly with 50 µL of 1xPBS, pDNA or self-amplifying RNA vectors (pDNA or RNA, 5 µg/site). Images were obtained 2 days following treatment and are shown as representative of 6–8 slides taken per area, n = 4 muscles per group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

vaccines-01-00367-f002: Histology of muscle tissues (transverse) by hematoxylin and eosin (H&E) staining, and expression of reporter protein (GFP) as delivered by intramuscular injection without EP (No EP) or with EP (+EP). Mice were injected intramuscularly with 50 µL of 1xPBS, pDNA or self-amplifying RNA vectors (pDNA or RNA, 5 µg/site). Images were obtained 2 days following treatment and are shown as representative of 6–8 slides taken per area, n = 4 muscles per group.
Mentions: To assess the effect of EP-enhanced delivery of nucleic acid vaccines in muscle, mice were injected intramuscularly with pDNA or self-amplifying mRNA (5 μg/site) with or without EP. The treated muscles were excised for GFP expression analysis and for hematoxylin and eosin (H&E) staining. Representative images of GFP expression in mouse muscle are shown in Figure 2. Robust GFP expression was detected in both pDNA + EP and RNA + EP treated groups. Low levels of GFP expression were detected in the RNA (no EP) control group, but not in the pDNA (no EP) or 1× PBS control groups. These results are consistent with that observed in the SEAP studies.

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.