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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

Stability of rBCG-CMX in vivo.(A) Images of plates showing the mycobacterial growth of rBCG-CMX recovered from the dorsal region of mice 5, 10 and 15 days after subcutaneous immunization, and plated on media with kanamycin (kan) or without (W/o). (B) CFU counts recovered at different time points from the dorsal region of mice after immunization. (C) CMX gene detection by PCR for three isolated colonies from plates W/o kanamycin (Lanes 3–5). Lanes 1: M: molecular weight marker; 2: Negative control: NC: water.
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pone-0112848-g002: Stability of rBCG-CMX in vivo.(A) Images of plates showing the mycobacterial growth of rBCG-CMX recovered from the dorsal region of mice 5, 10 and 15 days after subcutaneous immunization, and plated on media with kanamycin (kan) or without (W/o). (B) CFU counts recovered at different time points from the dorsal region of mice after immunization. (C) CMX gene detection by PCR for three isolated colonies from plates W/o kanamycin (Lanes 3–5). Lanes 1: M: molecular weight marker; 2: Negative control: NC: water.

Mentions: In order to verify the stability of the plasmid within the recombinant vaccine rBCG-CMX invivo without antibiotic selective pressure, mice were vaccinated subcutaneously and the tissue of the site of infection was macerated at different time points and plated on media with or without the selective antibiotic kanamycin. As shown in Figure 2, the number of CFU recovered from media with or without antibiotic was similar, indicating that the recombinant vaccine recovered from mice retained the plasmids up to 15 days after immunization (Figs. 2A and B). The presence of plasmid was further confirmed by performing PCR specific to the CMX gene (Fig. 2C).


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)

Stability of rBCG-CMX in vivo.(A) Images of plates showing the mycobacterial growth of rBCG-CMX recovered from the dorsal region of mice 5, 10 and 15 days after subcutaneous immunization, and plated on media with kanamycin (kan) or without (W/o). (B) CFU counts recovered at different time points from the dorsal region of mice after immunization. (C) CMX gene detection by PCR for three isolated colonies from plates W/o kanamycin (Lanes 3–5). Lanes 1: M: molecular weight marker; 2: Negative control: NC: water.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0112848-g002: Stability of rBCG-CMX in vivo.(A) Images of plates showing the mycobacterial growth of rBCG-CMX recovered from the dorsal region of mice 5, 10 and 15 days after subcutaneous immunization, and plated on media with kanamycin (kan) or without (W/o). (B) CFU counts recovered at different time points from the dorsal region of mice after immunization. (C) CMX gene detection by PCR for three isolated colonies from plates W/o kanamycin (Lanes 3–5). Lanes 1: M: molecular weight marker; 2: Negative control: NC: water.
Mentions: In order to verify the stability of the plasmid within the recombinant vaccine rBCG-CMX invivo without antibiotic selective pressure, mice were vaccinated subcutaneously and the tissue of the site of infection was macerated at different time points and plated on media with or without the selective antibiotic kanamycin. As shown in Figure 2, the number of CFU recovered from media with or without antibiotic was similar, indicating that the recombinant vaccine recovered from mice retained the plasmids up to 15 days after immunization (Figs. 2A and B). The presence of plasmid was further confirmed by performing PCR specific to the CMX gene (Fig. 2C).

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