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Recombinant HBHA boosting effect on BCG-induced immunity against Mycobacterium tuberculosis infection.

Guerrero GG, Locht C - Clin. Dev. Immunol. (2011)

Bottom Line: However, nHBHA is characterized by a complex methylation pattern in its C-terminal domain, which is important for protective immunogenicity in primary vaccination.We found that while subcutaneous rHBHA boosting enhanced protection of BCG-primed mice against intranasal M. tuberculosis infection both in spleen and lungs, enhanced protection against aerosol infection was only seen in the spleen (0.72 logs; P < 0.05) but not in the lungs.This report thus provides evidence that rHBHA may be considered as a booster vaccine against disseminated tuberculosis.

View Article: PubMed Central - PubMed

Affiliation: INSERM U1019, Lille, France. gloriaguillermina@correo.unam.mx

ABSTRACT
Heterologous prime-boost regimens are effective strategies to promote long-term memory and strong cellular Th1 responses to Mycobacterium tuberculosis, when BCG is used in the priming step. Subcutaneous or intranasal boosting of BCG-vaccinated newborn mice with native heparin-binding haemagglutinin (nHBHA) significantly enhances protection against M. tuberculosis. However, nHBHA is characterized by a complex methylation pattern in its C-terminal domain, which is important for protective immunogenicity in primary vaccination. In this study we addressed the question whether boosting with recombinant, non-methylated HBHA (rHBHA) produced in Escherichia coli may enhance protection of BCG-primed newborn mice. We found that while subcutaneous rHBHA boosting enhanced protection of BCG-primed mice against intranasal M. tuberculosis infection both in spleen and lungs, enhanced protection against aerosol infection was only seen in the spleen (0.72 logs; P < 0.05) but not in the lungs. Thus, in BCG-primed mice the methylation of the C-terminal domain of HBHA is dispensable for the induction of enhanced protection in the lungs against intranasal but not aerosol infection, whereas it enhances protection in the spleen in both challenge models. This report thus provides evidence that rHBHA may be considered as a booster vaccine against disseminated tuberculosis.

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Related in: MedlinePlus

Spleen cell cytokine profile of BCG-vaccinated mice with or without rHBHA boost after M. tuberculosis infection. Eight weeks after i.n. (A) or aerosol (B) challenge spleen cells from control mice (white bars), BCG-vaccinated mice (black bars), or BCG-vaccinated/rHBHA-boosted mice (gray bars) were cultured in the presence of medium only, ConA (2.5 μg/mL), or rHBHA (5 μg/mL). Levels of cytokines were measured after 72 h culture in the supernatants by using the OptEIA kit (BD Biosciences). Values are expressed in pg/mL and represent media ± SEM of samples tested in duplicates from each group of mice. The data was considered statistically significant when P < 0.05 (*), P < 0.001 (**), and P < 0.0001 (***).
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fig1: Spleen cell cytokine profile of BCG-vaccinated mice with or without rHBHA boost after M. tuberculosis infection. Eight weeks after i.n. (A) or aerosol (B) challenge spleen cells from control mice (white bars), BCG-vaccinated mice (black bars), or BCG-vaccinated/rHBHA-boosted mice (gray bars) were cultured in the presence of medium only, ConA (2.5 μg/mL), or rHBHA (5 μg/mL). Levels of cytokines were measured after 72 h culture in the supernatants by using the OptEIA kit (BD Biosciences). Values are expressed in pg/mL and represent media ± SEM of samples tested in duplicates from each group of mice. The data was considered statistically significant when P < 0.05 (*), P < 0.001 (**), and P < 0.0001 (***).

Mentions: Splenocytes from i.n. infected mice after in vitro recall with rHBHA produced significant amounts of IL12p70 (3325 ± 403 pg/mL) (Figure 1A, gray bar), in comparison with PBS-immunized mice (850 ± 508 pg/mL) (Figure 1A, white bar) and BCG-vaccinated non-boosted mice (1540 ± 239 pg/mL) (Figure 1A, black bar). In contrast, the spleen cell TGF-β production in BCG-vaccinated/rHBHA-boosted mice after infection was similar (2174 ± 270 pg/mL) to that of spleen cells from the BCG-vaccinated i.n. infected mice without boost (2291 ± 203 pg/mL) (Figure 1A). BCG-vaccinated/rHBHA-boosted mice also produced slightly higher amounts of IFN-γ (505 ± 150 pg/mL, P < 0.05) (Figure 1A) after infection, in comparison with those induced by the BCG-vaccinated non-boosted mice (252 ± 14 pg/mL) (Figure 1A) and PBS control mice (236 ± 32) (Figure 1A). We also determined regulatory (IL-10) and Th17 type cytokines (IL-6, IL-17) and found that splenocytes from BCG-vaccinated/rHBHA-boosted i.n. infected mice induced a significant IL-10 production (1640 ± 100 pg/mL) in comparison with infected PBS control mice (76 ± 20 pg/mL) and infected BCG-vaccinated mice without boost (304 ± 14 pg/mL) (Figure 1A). Interestingly, the IL-6 spleen cell production from BCG-vaccinated/rHBHA-boosted i.n. infected mice (427 ± 89 pg/mL) was similar to that of the non-boosted group (419 ± 20 pg/mL), but significantly higher than that of the PBS control mice (96 ± 4 pg/mL) (Figure 1A). Spleen cell IL-17 production significantly changed between the three groups after i.n. infection (Figure 1A).


Recombinant HBHA boosting effect on BCG-induced immunity against Mycobacterium tuberculosis infection.

Guerrero GG, Locht C - Clin. Dev. Immunol. (2011)

Spleen cell cytokine profile of BCG-vaccinated mice with or without rHBHA boost after M. tuberculosis infection. Eight weeks after i.n. (A) or aerosol (B) challenge spleen cells from control mice (white bars), BCG-vaccinated mice (black bars), or BCG-vaccinated/rHBHA-boosted mice (gray bars) were cultured in the presence of medium only, ConA (2.5 μg/mL), or rHBHA (5 μg/mL). Levels of cytokines were measured after 72 h culture in the supernatants by using the OptEIA kit (BD Biosciences). Values are expressed in pg/mL and represent media ± SEM of samples tested in duplicates from each group of mice. The data was considered statistically significant when P < 0.05 (*), P < 0.001 (**), and P < 0.0001 (***).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3102518&req=5

fig1: Spleen cell cytokine profile of BCG-vaccinated mice with or without rHBHA boost after M. tuberculosis infection. Eight weeks after i.n. (A) or aerosol (B) challenge spleen cells from control mice (white bars), BCG-vaccinated mice (black bars), or BCG-vaccinated/rHBHA-boosted mice (gray bars) were cultured in the presence of medium only, ConA (2.5 μg/mL), or rHBHA (5 μg/mL). Levels of cytokines were measured after 72 h culture in the supernatants by using the OptEIA kit (BD Biosciences). Values are expressed in pg/mL and represent media ± SEM of samples tested in duplicates from each group of mice. The data was considered statistically significant when P < 0.05 (*), P < 0.001 (**), and P < 0.0001 (***).
Mentions: Splenocytes from i.n. infected mice after in vitro recall with rHBHA produced significant amounts of IL12p70 (3325 ± 403 pg/mL) (Figure 1A, gray bar), in comparison with PBS-immunized mice (850 ± 508 pg/mL) (Figure 1A, white bar) and BCG-vaccinated non-boosted mice (1540 ± 239 pg/mL) (Figure 1A, black bar). In contrast, the spleen cell TGF-β production in BCG-vaccinated/rHBHA-boosted mice after infection was similar (2174 ± 270 pg/mL) to that of spleen cells from the BCG-vaccinated i.n. infected mice without boost (2291 ± 203 pg/mL) (Figure 1A). BCG-vaccinated/rHBHA-boosted mice also produced slightly higher amounts of IFN-γ (505 ± 150 pg/mL, P < 0.05) (Figure 1A) after infection, in comparison with those induced by the BCG-vaccinated non-boosted mice (252 ± 14 pg/mL) (Figure 1A) and PBS control mice (236 ± 32) (Figure 1A). We also determined regulatory (IL-10) and Th17 type cytokines (IL-6, IL-17) and found that splenocytes from BCG-vaccinated/rHBHA-boosted i.n. infected mice induced a significant IL-10 production (1640 ± 100 pg/mL) in comparison with infected PBS control mice (76 ± 20 pg/mL) and infected BCG-vaccinated mice without boost (304 ± 14 pg/mL) (Figure 1A). Interestingly, the IL-6 spleen cell production from BCG-vaccinated/rHBHA-boosted i.n. infected mice (427 ± 89 pg/mL) was similar to that of the non-boosted group (419 ± 20 pg/mL), but significantly higher than that of the PBS control mice (96 ± 4 pg/mL) (Figure 1A). Spleen cell IL-17 production significantly changed between the three groups after i.n. infection (Figure 1A).

Bottom Line: However, nHBHA is characterized by a complex methylation pattern in its C-terminal domain, which is important for protective immunogenicity in primary vaccination.We found that while subcutaneous rHBHA boosting enhanced protection of BCG-primed mice against intranasal M. tuberculosis infection both in spleen and lungs, enhanced protection against aerosol infection was only seen in the spleen (0.72 logs; P < 0.05) but not in the lungs.This report thus provides evidence that rHBHA may be considered as a booster vaccine against disseminated tuberculosis.

View Article: PubMed Central - PubMed

Affiliation: INSERM U1019, Lille, France. gloriaguillermina@correo.unam.mx

ABSTRACT
Heterologous prime-boost regimens are effective strategies to promote long-term memory and strong cellular Th1 responses to Mycobacterium tuberculosis, when BCG is used in the priming step. Subcutaneous or intranasal boosting of BCG-vaccinated newborn mice with native heparin-binding haemagglutinin (nHBHA) significantly enhances protection against M. tuberculosis. However, nHBHA is characterized by a complex methylation pattern in its C-terminal domain, which is important for protective immunogenicity in primary vaccination. In this study we addressed the question whether boosting with recombinant, non-methylated HBHA (rHBHA) produced in Escherichia coli may enhance protection of BCG-primed newborn mice. We found that while subcutaneous rHBHA boosting enhanced protection of BCG-primed mice against intranasal M. tuberculosis infection both in spleen and lungs, enhanced protection against aerosol infection was only seen in the spleen (0.72 logs; P < 0.05) but not in the lungs. Thus, in BCG-primed mice the methylation of the C-terminal domain of HBHA is dispensable for the induction of enhanced protection in the lungs against intranasal but not aerosol infection, whereas it enhances protection in the spleen in both challenge models. This report thus provides evidence that rHBHA may be considered as a booster vaccine against disseminated tuberculosis.

Show MeSH
Related in: MedlinePlus