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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

Secretion of IFN-γ (a) and IL-10 (b) by human naive (grey bars) and memory (black bars) T cells following 24 h and 96 h coculture with PPD- (10 μg/mL), BCG- (1 : 1), rBCGIL-18- (1 : 1), or LPS- (1 μg/mL) pulsed autologous MoDCs (ratio MoDCs/T cells, 1 : 10). The cytokine levels in the cocultures were measured by ELISA. Data shown are the mean ± SEM of 20 independent donors. Statistical analyses were performed using the Kruskal-Wallis test and Mann-Whitney U test. *P < 0.05; **P < 0.01.
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fig7: Secretion of IFN-γ (a) and IL-10 (b) by human naive (grey bars) and memory (black bars) T cells following 24 h and 96 h coculture with PPD- (10 μg/mL), BCG- (1 : 1), rBCGIL-18- (1 : 1), or LPS- (1 μg/mL) pulsed autologous MoDCs (ratio MoDCs/T cells, 1 : 10). The cytokine levels in the cocultures were measured by ELISA. Data shown are the mean ± SEM of 20 independent donors. Statistical analyses were performed using the Kruskal-Wallis test and Mann-Whitney U test. *P < 0.05; **P < 0.01.

Mentions: In order to investigate whether rBCGhIL-18-pulsed DC can polarize T cells towards the Th1 profile, we analyzed the cytokine responses of the autologous naive and memory CD4+ T cells upon incubation with DCs stimulated with PPD, BCG, or rBCGhIL-18. As shown in Figure 7(a), naive T cells produced significantly more IFN-γ in response to rBCGhIL-18-stimulated DC coincubation, as compared to T cells incubation with DCs stimulated with PPD or BCG, both after 24 h and after 96 h of culture. PPD- or BCG-stimulated DCs induced only low levels of IFN-γ by the naive T cells. Similarly, rBCGhIL-18-pulsed DC also stimulated the secretion of IL-10 by naive T cells, whereas this was much less the case for PPD- or BCG-pulsed DCs (Figure 7(b)) In contrast to the naive T cells, memory T cells produced IFN-γ upon coculture with BCG-, rBCGhIL-18-, or PPD-pulsed DC at comparable levels. Similar results were obtained for the production of IL-10 by the T cells. Interestingly, both IFN-γ and IL-10 were produced at higher levels by naive T cells upon incubation with rBCGhIL-18-pulsed DC, compared to memory T, whereas the reverse was seen for IFN-γ upon coculture with PPD- or BCG-pulsed DCs. There were no differences in the IL-10 production by naive and memory T cells in response to PPD-pulsed MoDCs.


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)

Secretion of IFN-γ (a) and IL-10 (b) by human naive (grey bars) and memory (black bars) T cells following 24 h and 96 h coculture with PPD- (10 μg/mL), BCG- (1 : 1), rBCGIL-18- (1 : 1), or LPS- (1 μg/mL) pulsed autologous MoDCs (ratio MoDCs/T cells, 1 : 10). The cytokine levels in the cocultures were measured by ELISA. Data shown are the mean ± SEM of 20 independent donors. Statistical analyses were performed using the Kruskal-Wallis test and Mann-Whitney U test. *P < 0.05; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig7: Secretion of IFN-γ (a) and IL-10 (b) by human naive (grey bars) and memory (black bars) T cells following 24 h and 96 h coculture with PPD- (10 μg/mL), BCG- (1 : 1), rBCGIL-18- (1 : 1), or LPS- (1 μg/mL) pulsed autologous MoDCs (ratio MoDCs/T cells, 1 : 10). The cytokine levels in the cocultures were measured by ELISA. Data shown are the mean ± SEM of 20 independent donors. Statistical analyses were performed using the Kruskal-Wallis test and Mann-Whitney U test. *P < 0.05; **P < 0.01.
Mentions: In order to investigate whether rBCGhIL-18-pulsed DC can polarize T cells towards the Th1 profile, we analyzed the cytokine responses of the autologous naive and memory CD4+ T cells upon incubation with DCs stimulated with PPD, BCG, or rBCGhIL-18. As shown in Figure 7(a), naive T cells produced significantly more IFN-γ in response to rBCGhIL-18-stimulated DC coincubation, as compared to T cells incubation with DCs stimulated with PPD or BCG, both after 24 h and after 96 h of culture. PPD- or BCG-stimulated DCs induced only low levels of IFN-γ by the naive T cells. Similarly, rBCGhIL-18-pulsed DC also stimulated the secretion of IL-10 by naive T cells, whereas this was much less the case for PPD- or BCG-pulsed DCs (Figure 7(b)) In contrast to the naive T cells, memory T cells produced IFN-γ upon coculture with BCG-, rBCGhIL-18-, or PPD-pulsed DC at comparable levels. Similar results were obtained for the production of IL-10 by the T cells. Interestingly, both IFN-γ and IL-10 were produced at higher levels by naive T cells upon incubation with rBCGhIL-18-pulsed DC, compared to memory T, whereas the reverse was seen for IFN-γ upon coculture with PPD- or BCG-pulsed DCs. There were no differences in the IL-10 production by naive and memory T cells in response to PPD-pulsed MoDCs.

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