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Engineering new mycobacterial vaccine design for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG.

Saubi N, Gea-Mallorquí E, Ferrer P, Hurtado C, Sánchez-Úbeda S, Eto Y, Gatell JM, Hanke T, Joseph J - Mol Ther Methods Clin Dev (2014)

Bottom Line: HIVA was safe and induced HIV-1 and Mycobacterium tuberculosis-specific interferon-γ-producing T-cell responses in adult BALB/c mice.Thus, we engineered a novel, safer, good laboratory practice-compatible BCG-vectored vaccine using prototype immunogen HIVA.This antibiotic-free plasmid selection system based on "double" auxotrophic complementation might be a new mycobacterial vaccine platform to develop not only recombinant BCG-based vaccines expressing second generation of HIV-1 immunogens but also other major pediatric pathogens to prime protective response soon after birth.

View Article: PubMed Central - PubMed

Affiliation: AIDS Research Group, Hospital Clinic/HIVACAT, School of Medicine, University of Barcelona , Barcelona, Catalonia, Spain.

ABSTRACT
In this study, we have engineered a new mycobacterial vaccine design by using an antibiotic-free plasmid selection system. We assembled a novel Escherichia coli (E. coli)-mycobacterial shuttle plasmid p2auxo.HIVA, expressing the HIV-1 clade A immunogen HIVA. This shuttle vector employs an antibiotic resistance-free mechanism for plasmid selection and maintenance based on glycine complementation in E. coli and lysine complementation in mycobacteria. This plasmid was first transformed into glycine auxotroph of E. coli strain and subsequently transformed into lysine auxotroph of Mycobacterium bovis BCG strain to generate vaccine BCG.HIVA(2auxo). We demonstrated that the episomal plasmid p2auxo.HIVA was stable in vivo over a 7-week period and genetically and phenotypically characterized the BCG.HIVA(2auxo) vaccine strain. The BCG.HIVA(2auxo) vaccine in combination with modified vaccinia virus Ankara (MVA). HIVA was safe and induced HIV-1 and Mycobacterium tuberculosis-specific interferon-γ-producing T-cell responses in adult BALB/c mice. Polyfunctional HIV-1-specific CD8+ T cells, which produce interferon-γ and tumor necrosis factor-α and express the degranulation marker CD107a, were induced. Thus, we engineered a novel, safer, good laboratory practice-compatible BCG-vectored vaccine using prototype immunogen HIVA. This antibiotic-free plasmid selection system based on "double" auxotrophic complementation might be a new mycobacterial vaccine platform to develop not only recombinant BCG-based vaccines expressing second generation of HIV-1 immunogens but also other major pediatric pathogens to prime protective response soon after birth.

No MeSH data available.


Related in: MedlinePlus

BCG.HIVA2auxo prime and MVA.HIVA boost safety in adult mice. (a) Adult mice were either left unimmunized or immunized with 106 cfu of BCG wild type or BCG.HIVA2auxo by intradermal route and subsequently given a booster dose of 106 pfu of MVA.HIVA at week 12 by intramuscular route. (b) The body weight was recorded over time, and the mean for each group of mice is shown (n = 10 for group 1 and n = 5 for groups 2, 3, and 4). Data from naive mice are presented as mean ± 2 SD (n = 5, dashed lines). The weight differences between vaccinated and naive mice group were analyzed in all monitored time points by analysis of variance test.
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fig5: BCG.HIVA2auxo prime and MVA.HIVA boost safety in adult mice. (a) Adult mice were either left unimmunized or immunized with 106 cfu of BCG wild type or BCG.HIVA2auxo by intradermal route and subsequently given a booster dose of 106 pfu of MVA.HIVA at week 12 by intramuscular route. (b) The body weight was recorded over time, and the mean for each group of mice is shown (n = 10 for group 1 and n = 5 for groups 2, 3, and 4). Data from naive mice are presented as mean ± 2 SD (n = 5, dashed lines). The weight differences between vaccinated and naive mice group were analyzed in all monitored time points by analysis of variance test.

Mentions: As shown in Figure 5b, the body mass was monitored over time and recorded to depict any adverse events and body mass loss due to vaccination. In order to detect vaccine-derived adverse events, a 12-week period between BCG-prime and MVA boost was established for this trial (Figure 5a). Importantly, no statistically significant difference was observed between the vaccinated mice groups and the naive mice group in all monitored time points. Furthermore, between weeks 1 and 14, the body mass monitored in all vaccinated mice groups was found to lie between the mean ± 2 SD body mass curve in naive mice (Figure 5b). It is also important to mention that no mice died during the trial, and no local adverse events and associated systemic reactions were observed.


Engineering new mycobacterial vaccine design for HIV-TB pediatric vaccine vectored by lysine auxotroph of BCG.

Saubi N, Gea-Mallorquí E, Ferrer P, Hurtado C, Sánchez-Úbeda S, Eto Y, Gatell JM, Hanke T, Joseph J - Mol Ther Methods Clin Dev (2014)

BCG.HIVA2auxo prime and MVA.HIVA boost safety in adult mice. (a) Adult mice were either left unimmunized or immunized with 106 cfu of BCG wild type or BCG.HIVA2auxo by intradermal route and subsequently given a booster dose of 106 pfu of MVA.HIVA at week 12 by intramuscular route. (b) The body weight was recorded over time, and the mean for each group of mice is shown (n = 10 for group 1 and n = 5 for groups 2, 3, and 4). Data from naive mice are presented as mean ± 2 SD (n = 5, dashed lines). The weight differences between vaccinated and naive mice group were analyzed in all monitored time points by analysis of variance test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362382&req=5

fig5: BCG.HIVA2auxo prime and MVA.HIVA boost safety in adult mice. (a) Adult mice were either left unimmunized or immunized with 106 cfu of BCG wild type or BCG.HIVA2auxo by intradermal route and subsequently given a booster dose of 106 pfu of MVA.HIVA at week 12 by intramuscular route. (b) The body weight was recorded over time, and the mean for each group of mice is shown (n = 10 for group 1 and n = 5 for groups 2, 3, and 4). Data from naive mice are presented as mean ± 2 SD (n = 5, dashed lines). The weight differences between vaccinated and naive mice group were analyzed in all monitored time points by analysis of variance test.
Mentions: As shown in Figure 5b, the body mass was monitored over time and recorded to depict any adverse events and body mass loss due to vaccination. In order to detect vaccine-derived adverse events, a 12-week period between BCG-prime and MVA boost was established for this trial (Figure 5a). Importantly, no statistically significant difference was observed between the vaccinated mice groups and the naive mice group in all monitored time points. Furthermore, between weeks 1 and 14, the body mass monitored in all vaccinated mice groups was found to lie between the mean ± 2 SD body mass curve in naive mice (Figure 5b). It is also important to mention that no mice died during the trial, and no local adverse events and associated systemic reactions were observed.

Bottom Line: HIVA was safe and induced HIV-1 and Mycobacterium tuberculosis-specific interferon-γ-producing T-cell responses in adult BALB/c mice.Thus, we engineered a novel, safer, good laboratory practice-compatible BCG-vectored vaccine using prototype immunogen HIVA.This antibiotic-free plasmid selection system based on "double" auxotrophic complementation might be a new mycobacterial vaccine platform to develop not only recombinant BCG-based vaccines expressing second generation of HIV-1 immunogens but also other major pediatric pathogens to prime protective response soon after birth.

View Article: PubMed Central - PubMed

Affiliation: AIDS Research Group, Hospital Clinic/HIVACAT, School of Medicine, University of Barcelona , Barcelona, Catalonia, Spain.

ABSTRACT
In this study, we have engineered a new mycobacterial vaccine design by using an antibiotic-free plasmid selection system. We assembled a novel Escherichia coli (E. coli)-mycobacterial shuttle plasmid p2auxo.HIVA, expressing the HIV-1 clade A immunogen HIVA. This shuttle vector employs an antibiotic resistance-free mechanism for plasmid selection and maintenance based on glycine complementation in E. coli and lysine complementation in mycobacteria. This plasmid was first transformed into glycine auxotroph of E. coli strain and subsequently transformed into lysine auxotroph of Mycobacterium bovis BCG strain to generate vaccine BCG.HIVA(2auxo). We demonstrated that the episomal plasmid p2auxo.HIVA was stable in vivo over a 7-week period and genetically and phenotypically characterized the BCG.HIVA(2auxo) vaccine strain. The BCG.HIVA(2auxo) vaccine in combination with modified vaccinia virus Ankara (MVA). HIVA was safe and induced HIV-1 and Mycobacterium tuberculosis-specific interferon-γ-producing T-cell responses in adult BALB/c mice. Polyfunctional HIV-1-specific CD8+ T cells, which produce interferon-γ and tumor necrosis factor-α and express the degranulation marker CD107a, were induced. Thus, we engineered a novel, safer, good laboratory practice-compatible BCG-vectored vaccine using prototype immunogen HIVA. This antibiotic-free plasmid selection system based on "double" auxotrophic complementation might be a new mycobacterial vaccine platform to develop not only recombinant BCG-based vaccines expressing second generation of HIV-1 immunogens but also other major pediatric pathogens to prime protective response soon after birth.

No MeSH data available.


Related in: MedlinePlus