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A new recombinant BCG vaccine induces specific Th17 and Th1 effector cells with higher protective efficacy against tuberculosis.

da Costa AC, Costa-Júnior Ade O, de Oliveira FM, Nogueira SV, Rosa JD, Resende DP, Kipnis A, Junqueira-Kipnis AP - PLoS ONE (2014)

Bottom Line: Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG.Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load.In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunopatologia das Doenças Infecciosas, Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, Goiás, Brazil.

ABSTRACT
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) that is a major public health problem. The vaccine used for TB prevention is Mycobacterium bovis bacillus Calmette-Guérin (BCG), which provides variable efficacy in protecting against pulmonary TB among adults. Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG. Here we constructed a new recombinant BCG (rBCG) vaccine expressing a fusion protein (CMX) composed of immune dominant epitopes from Ag85C, MPT51, and HspX and evaluated its immunogenicity and protection in a murine model of infection. The stability of the vaccine in vivo was maintained for up to 20 days post-vaccination. rBCG-CMX was efficiently phagocytized by peritoneal macrophages and induced nitric oxide (NO) production. Following mouse immunization, this vaccine induced a specific immune response in cells from lungs and spleen to the fusion protein and to each of the component recombinant proteins by themselves. Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load. In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination. This study describes the creation of a new promising vaccine for TB that we hope will be used in further studies to address its safety before proceeding to clinical trials.

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Related in: MedlinePlus

Levels of polyfunctional CD4+ T cells induced by BCG and rBCG-CMX vaccines.Spleen (A and C) and lung (B and D) cell suspensions from vaccinated and control mice stimulated with rCMX. (A–B) CD4+IL-2+IFN-γ+ cells or (C–D) CD4+TNF-α+IFN-γ+ cells were analyzed by flow cytometry. Lymphocytes were selected based on size and granularity. Gates were set to analyze CD4+ T cells, and then the fluorescence of antibodies detecting IL-2+ and IFN-γ+ or TNF-α+ and IFN-γ+ cells was recorded. These data are representative of two independent experiments (N = 6, *p<0.05).
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pone-0112848-g007: Levels of polyfunctional CD4+ T cells induced by BCG and rBCG-CMX vaccines.Spleen (A and C) and lung (B and D) cell suspensions from vaccinated and control mice stimulated with rCMX. (A–B) CD4+IL-2+IFN-γ+ cells or (C–D) CD4+TNF-α+IFN-γ+ cells were analyzed by flow cytometry. Lymphocytes were selected based on size and granularity. Gates were set to analyze CD4+ T cells, and then the fluorescence of antibodies detecting IL-2+ and IFN-γ+ or TNF-α+ and IFN-γ+ cells was recorded. These data are representative of two independent experiments (N = 6, *p<0.05).

Mentions: Although the above experiments determined that the rBCG-CMX vaccine generates Th1 (IFN-γ) and Th17 specific responses, it remained important to verify the induction of polyfunctional CD4+ T cells, since several publications have associated these cells with protection against Mtb [33], [37]. Ex vivo stimulation of spleen and lung cells with CMX increased the numbers of CD4+ T cells positive for both IL-2 and IFN-γ (Figs. 7A and C) as well as for both TNF-α and IFN-γ (Figs. 7B and D) in cells from rBCG-CMX vaccinated mice as compared to the levels in cells from BCG Moreau vaccinated mice.


A new recombinant BCG vaccine induces specific Th17 and Th1 effector cells with higher protective efficacy against tuberculosis.

da Costa AC, Costa-Júnior Ade O, de Oliveira FM, Nogueira SV, Rosa JD, Resende DP, Kipnis A, Junqueira-Kipnis AP - PLoS ONE (2014)

Levels of polyfunctional CD4+ T cells induced by BCG and rBCG-CMX vaccines.Spleen (A and C) and lung (B and D) cell suspensions from vaccinated and control mice stimulated with rCMX. (A–B) CD4+IL-2+IFN-γ+ cells or (C–D) CD4+TNF-α+IFN-γ+ cells were analyzed by flow cytometry. Lymphocytes were selected based on size and granularity. Gates were set to analyze CD4+ T cells, and then the fluorescence of antibodies detecting IL-2+ and IFN-γ+ or TNF-α+ and IFN-γ+ cells was recorded. These data are representative of two independent experiments (N = 6, *p<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232451&req=5

pone-0112848-g007: Levels of polyfunctional CD4+ T cells induced by BCG and rBCG-CMX vaccines.Spleen (A and C) and lung (B and D) cell suspensions from vaccinated and control mice stimulated with rCMX. (A–B) CD4+IL-2+IFN-γ+ cells or (C–D) CD4+TNF-α+IFN-γ+ cells were analyzed by flow cytometry. Lymphocytes were selected based on size and granularity. Gates were set to analyze CD4+ T cells, and then the fluorescence of antibodies detecting IL-2+ and IFN-γ+ or TNF-α+ and IFN-γ+ cells was recorded. These data are representative of two independent experiments (N = 6, *p<0.05).
Mentions: Although the above experiments determined that the rBCG-CMX vaccine generates Th1 (IFN-γ) and Th17 specific responses, it remained important to verify the induction of polyfunctional CD4+ T cells, since several publications have associated these cells with protection against Mtb [33], [37]. Ex vivo stimulation of spleen and lung cells with CMX increased the numbers of CD4+ T cells positive for both IL-2 and IFN-γ (Figs. 7A and C) as well as for both TNF-α and IFN-γ (Figs. 7B and D) in cells from rBCG-CMX vaccinated mice as compared to the levels in cells from BCG Moreau vaccinated mice.

Bottom Line: Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG.Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load.In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunopatologia das Doenças Infecciosas, Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, Goiás, Brazil.

ABSTRACT
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) that is a major public health problem. The vaccine used for TB prevention is Mycobacterium bovis bacillus Calmette-Guérin (BCG), which provides variable efficacy in protecting against pulmonary TB among adults. Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG. Here we constructed a new recombinant BCG (rBCG) vaccine expressing a fusion protein (CMX) composed of immune dominant epitopes from Ag85C, MPT51, and HspX and evaluated its immunogenicity and protection in a murine model of infection. The stability of the vaccine in vivo was maintained for up to 20 days post-vaccination. rBCG-CMX was efficiently phagocytized by peritoneal macrophages and induced nitric oxide (NO) production. Following mouse immunization, this vaccine induced a specific immune response in cells from lungs and spleen to the fusion protein and to each of the component recombinant proteins by themselves. Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load. In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination. This study describes the creation of a new promising vaccine for TB that we hope will be used in further studies to address its safety before proceeding to clinical trials.

Show MeSH
Related in: MedlinePlus