Limits...
Dendritic Cell Activity Driven by Recombinant Mycobacterium bovis BCG Producing Human IL-18, in Healthy BCG Vaccinated Adults.

Szpakowski P, Biet F, Locht C, Paszkiewicz M, Rudnicka W, Druszczyńska M, Allain F, Fol M, Pestel J, Kowalewicz-Kulbat M - J Immunol Res (2015)

Bottom Line: We found that the costimulatory CD86 and CD80 molecules were significantly upregulated on rBCGhIL-18-infected DCs, whereas the stimulation of DCs with nonrecombinant BCG was less effective.In contrast, both BCG strains decreased the DC-SIGN expression on human DCs.The rBCGhIL-18 increased IL-23, IL-10, and IP-10 production by DCs to a greater extent than nonrecombinant BCG.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, University of Lodz, Banacha Street 12/19, 90-237 Lodz, Poland.

ABSTRACT
Tuberculosis remains an enormous global burden, despite wide vaccination coverage with the Bacillus Calmette-Guérin (BCG), the only vaccine available against this disease, indicating that BCG-driven immunity is insufficient to protect the human population against tuberculosis. In this study we constructed recombinant BCG producing human IL-18 (rBCGhIL-18) and investigated whether human IL-18 produced by rBCGhIL-18 modulates DC functions and enhances Th1 responses to mycobacterial antigens in humans. We found that the costimulatory CD86 and CD80 molecules were significantly upregulated on rBCGhIL-18-infected DCs, whereas the stimulation of DCs with nonrecombinant BCG was less effective. In contrast, both BCG strains decreased the DC-SIGN expression on human DCs. The rBCGhIL-18 increased IL-23, IL-10, and IP-10 production by DCs to a greater extent than nonrecombinant BCG. In a coculture system of CD4(+) T cells and loaded DCs, rBCGhIL-18 favoured strong IFN-γ but also IL-10 production by naive T cells but not by memory T cells. This was much less the case for nonrecombinant BCG. Thus the expression of IL-18 by recombinant BCG increases IL-23, IP-10, and IL-10 expression by human DCs and enhances their ability to induce IFN-γ and IL-10 expression by naive T cells, without affecting the maturation phenotype of the DCs.

No MeSH data available.


Related in: MedlinePlus

IL-12p70, IL-10, IL-23, and IP-10 production by stimulated MoDCs. Human MoDCs were stimulated with PPD (10 μg/mL), BCG (1 : 1), rBCGIL-18 (1 : 1), or LPS (1 μg/mL) for 24 h or were left untreated (DC). The cytokine and chemokine levels in the culture were measured by ELISA. Data shown are the mean ± SEM of 40 independent donors. Statistical analyses were performed using the Kruskal-Wallis test. *P < 0.05; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4539176&req=5

fig6: IL-12p70, IL-10, IL-23, and IP-10 production by stimulated MoDCs. Human MoDCs were stimulated with PPD (10 μg/mL), BCG (1 : 1), rBCGIL-18 (1 : 1), or LPS (1 μg/mL) for 24 h or were left untreated (DC). The cytokine and chemokine levels in the culture were measured by ELISA. Data shown are the mean ± SEM of 40 independent donors. Statistical analyses were performed using the Kruskal-Wallis test. *P < 0.05; **P < 0.01.

Mentions: Next we analyzed the cytokine profile of the mycobacterial-pulsed DC that might affect the T-cell polarization. In particular, we analyzed IL-12, IL-23, and IL-10 in the supernatants of the DCs incubated with PPD, BCG, or rBCGhIL-18. In all blood donors, LPS-primed MoDCs produced significant levels of IL-12p70. In contrast, DCs infected with BCG and rBCGhIL-18 or stimulated with PPD, failed to secrete IL-12p70 in 6-hour cultures (data not shown) as well as in 24 hour cultures (Figure 6). Since bioactive IL-12 consists of p35 and p40 subunits, and the p40 subunit is also present in other IL-12 family members, such as IL-23, we determined the amount of secreted IL-23 in the supernatants of mycobacterial-stimulated DC cultures. Among the mycobacterial products, only rBCGhIL-18 induced significant IL-23 production compare to unpulsed DCs (P = 0.001), albeit much less than LPS (roughly 1/3 less). A trend was also seen for nonrecombinant BCG, but did not reach statistical significance, leading therefore to statistical significance between BCG and rBCGhIL-18 (Figure 6). In addition to IL-23, rBCGhIL-18 also induced the production of IL-10 by the DCs, although significant amounts of IL-10 were also produced upon stimulation with BCG, but not with PPD. However, rBCGhIL-18 induced significantly more IL-10 than BCG (P = 0.001), albeit again much less than LPS. Finally, rBCGhIL-18 also induced significant levels of IP-10 (CXCL10), known to be involved in Th1 cell recruitment, whereas this was not the case for PPD or nonrecombinant BCG, suggesting that rBCGhIL-18 might favor T cell towards a Th1 profile.


Dendritic Cell Activity Driven by Recombinant Mycobacterium bovis BCG Producing Human IL-18, in Healthy BCG Vaccinated Adults.

Szpakowski P, Biet F, Locht C, Paszkiewicz M, Rudnicka W, Druszczyńska M, Allain F, Fol M, Pestel J, Kowalewicz-Kulbat M - J Immunol Res (2015)

IL-12p70, IL-10, IL-23, and IP-10 production by stimulated MoDCs. Human MoDCs were stimulated with PPD (10 μg/mL), BCG (1 : 1), rBCGIL-18 (1 : 1), or LPS (1 μg/mL) for 24 h or were left untreated (DC). The cytokine and chemokine levels in the culture were measured by ELISA. Data shown are the mean ± SEM of 40 independent donors. Statistical analyses were performed using the Kruskal-Wallis test. *P < 0.05; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4539176&req=5

fig6: IL-12p70, IL-10, IL-23, and IP-10 production by stimulated MoDCs. Human MoDCs were stimulated with PPD (10 μg/mL), BCG (1 : 1), rBCGIL-18 (1 : 1), or LPS (1 μg/mL) for 24 h or were left untreated (DC). The cytokine and chemokine levels in the culture were measured by ELISA. Data shown are the mean ± SEM of 40 independent donors. Statistical analyses were performed using the Kruskal-Wallis test. *P < 0.05; **P < 0.01.
Mentions: Next we analyzed the cytokine profile of the mycobacterial-pulsed DC that might affect the T-cell polarization. In particular, we analyzed IL-12, IL-23, and IL-10 in the supernatants of the DCs incubated with PPD, BCG, or rBCGhIL-18. In all blood donors, LPS-primed MoDCs produced significant levels of IL-12p70. In contrast, DCs infected with BCG and rBCGhIL-18 or stimulated with PPD, failed to secrete IL-12p70 in 6-hour cultures (data not shown) as well as in 24 hour cultures (Figure 6). Since bioactive IL-12 consists of p35 and p40 subunits, and the p40 subunit is also present in other IL-12 family members, such as IL-23, we determined the amount of secreted IL-23 in the supernatants of mycobacterial-stimulated DC cultures. Among the mycobacterial products, only rBCGhIL-18 induced significant IL-23 production compare to unpulsed DCs (P = 0.001), albeit much less than LPS (roughly 1/3 less). A trend was also seen for nonrecombinant BCG, but did not reach statistical significance, leading therefore to statistical significance between BCG and rBCGhIL-18 (Figure 6). In addition to IL-23, rBCGhIL-18 also induced the production of IL-10 by the DCs, although significant amounts of IL-10 were also produced upon stimulation with BCG, but not with PPD. However, rBCGhIL-18 induced significantly more IL-10 than BCG (P = 0.001), albeit again much less than LPS. Finally, rBCGhIL-18 also induced significant levels of IP-10 (CXCL10), known to be involved in Th1 cell recruitment, whereas this was not the case for PPD or nonrecombinant BCG, suggesting that rBCGhIL-18 might favor T cell towards a Th1 profile.

Bottom Line: We found that the costimulatory CD86 and CD80 molecules were significantly upregulated on rBCGhIL-18-infected DCs, whereas the stimulation of DCs with nonrecombinant BCG was less effective.In contrast, both BCG strains decreased the DC-SIGN expression on human DCs.The rBCGhIL-18 increased IL-23, IL-10, and IP-10 production by DCs to a greater extent than nonrecombinant BCG.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, University of Lodz, Banacha Street 12/19, 90-237 Lodz, Poland.

ABSTRACT
Tuberculosis remains an enormous global burden, despite wide vaccination coverage with the Bacillus Calmette-Guérin (BCG), the only vaccine available against this disease, indicating that BCG-driven immunity is insufficient to protect the human population against tuberculosis. In this study we constructed recombinant BCG producing human IL-18 (rBCGhIL-18) and investigated whether human IL-18 produced by rBCGhIL-18 modulates DC functions and enhances Th1 responses to mycobacterial antigens in humans. We found that the costimulatory CD86 and CD80 molecules were significantly upregulated on rBCGhIL-18-infected DCs, whereas the stimulation of DCs with nonrecombinant BCG was less effective. In contrast, both BCG strains decreased the DC-SIGN expression on human DCs. The rBCGhIL-18 increased IL-23, IL-10, and IP-10 production by DCs to a greater extent than nonrecombinant BCG. In a coculture system of CD4(+) T cells and loaded DCs, rBCGhIL-18 favoured strong IFN-γ but also IL-10 production by naive T cells but not by memory T cells. This was much less the case for nonrecombinant BCG. Thus the expression of IL-18 by recombinant BCG increases IL-23, IP-10, and IL-10 expression by human DCs and enhances their ability to induce IFN-γ and IL-10 expression by naive T cells, without affecting the maturation phenotype of the DCs.

No MeSH data available.


Related in: MedlinePlus