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Human CD103(+) dendritic cells promote the differentiation of Porphyromonas gingivalis heat shock protein peptide-specific regulatory T cells.

Kim MJ, Jeong EK, Kwon EY, Joo JY, Lee JY, Choi J - J Periodontal Implant Sci (2014)

Bottom Line: Exogenous interleukin 2 was added as a coculture supplement.RA enhanced the conversion of CD103(+) DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance.RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, Pusan University School of Dentistry, Yangsan, Korea.

ABSTRACT

Purpose: Regulatory T cells (Tregs), expressing CD4 and CD25 as well as Foxp3, are known to play a pivotal role in immunoregulatory function in autoimmune diseases, cancers, and graft rejection. Dendritic cells (DCs) are considered the major antigen-presenting cells (APCs) for initiating these T-cell immune responses, of which CD103(+) DCs are derived from precursor human peripheral blood mononuclear cells (PBMCs). The aim of the present study was to evaluate the capacity of these PBMC-derived CD103(+) DCs to promote the differentiation of antigen-specific Tregs.

Methods: Monocyte-derived DCs were induced from CD14(+) monocytes from the PBMCs of 10 healthy subjects. Once the CD103(+) DCs were purified, the cell population was enriched by adding retinoic acid (RA). Peptide numbers 14 and 19 of Porphyromonas gingivalis heat shock protein 60 (HSP60) were synthesized to pulse CD103(+) DCs as a tool for presenting the peptide antigens to stimulate CD3(+) T cells that were isolated from human PBMC. Exogenous interleukin 2 was added as a coculture supplement. The antigen-specific T-cell lines established were phenotypically identified for their expression of CD4, CD25, or Foxp3.

Results: When PBMCs were used as APCs, they demonstrated only a marginal capacity to stimulate peptide-specific Tregs, whereas CD103(+) DCs showed a potent antigen presenting capability to promote the peptide-specific Tregs, especially for peptide 14. RA enhanced the conversion of CD103(+) DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance.

Conclusions: We demonstrated that CD103(+) DCs can promote antigen-specific Tregs from naive T cells, when used as APCs for an epitope peptide from P. gingivalis HSP60. RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

No MeSH data available.


Related in: MedlinePlus

Comparison of the effect of retinoic acid (RA) treatment and nontreatment on regulatory T cells (Tregs) induction by peptide number 14 from Porphyromonas gingivalis heat shock protein 60 (HSP60). (A) T cells were stained for CD4, CD25, and Foxp3 and analyzed by fluorescence activated cell sorter. The percentage of peptide-specific Tregs induced increased from 23.70% to 32.54% (subject 1 in C), and it increased from 27.57% to 34.83% (subject 2 in C) for peptide number 14 from P. gingivalis HSP60 by the addition of RA. (B) The bar graph shows the percentage of peptide-specific Tregs among T cells in two subjects. The difference between cases with and without RA failed to reach statistical significance. UL: upper left panel, UR: upper right panel, LL: lower left panel, LR: lower right panel.
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Figure 6: Comparison of the effect of retinoic acid (RA) treatment and nontreatment on regulatory T cells (Tregs) induction by peptide number 14 from Porphyromonas gingivalis heat shock protein 60 (HSP60). (A) T cells were stained for CD4, CD25, and Foxp3 and analyzed by fluorescence activated cell sorter. The percentage of peptide-specific Tregs induced increased from 23.70% to 32.54% (subject 1 in C), and it increased from 27.57% to 34.83% (subject 2 in C) for peptide number 14 from P. gingivalis HSP60 by the addition of RA. (B) The bar graph shows the percentage of peptide-specific Tregs among T cells in two subjects. The difference between cases with and without RA failed to reach statistical significance. UL: upper left panel, UR: upper right panel, LL: lower left panel, LR: lower right panel.

Mentions: RA has been reported to stimulate the expression of CD103 by DCs when added to the culture medium after culturing the DCs for 6 days. As shown in Fig. 4, the morphology of the DCs exhibited the typical extension of dendritic processes characteristic of maturing DCs. The CD103+ cells were increased from 4.28% to 32.67%. This characteristic phenotype expression occurred in parallel with its functional capability of inducing Tregs. After adding RA, the percentage of induced Tregs increased from 21.12% to 22.89% in subject 1, and from 27.73% to 34.46% in subject 2 for peptide number 19, respectively (Fig. 5). In case of peptide 14, it increased from 23.70% to 32.54% in subject 1 and from 27.57% to 34.83% in subject 2 (Fig. 6). There were no statistical differences in Tregs stimulated by DCs and RA-stimulated DCs for both peptide 19 and 14.


Human CD103(+) dendritic cells promote the differentiation of Porphyromonas gingivalis heat shock protein peptide-specific regulatory T cells.

Kim MJ, Jeong EK, Kwon EY, Joo JY, Lee JY, Choi J - J Periodontal Implant Sci (2014)

Comparison of the effect of retinoic acid (RA) treatment and nontreatment on regulatory T cells (Tregs) induction by peptide number 14 from Porphyromonas gingivalis heat shock protein 60 (HSP60). (A) T cells were stained for CD4, CD25, and Foxp3 and analyzed by fluorescence activated cell sorter. The percentage of peptide-specific Tregs induced increased from 23.70% to 32.54% (subject 1 in C), and it increased from 27.57% to 34.83% (subject 2 in C) for peptide number 14 from P. gingivalis HSP60 by the addition of RA. (B) The bar graph shows the percentage of peptide-specific Tregs among T cells in two subjects. The difference between cases with and without RA failed to reach statistical significance. UL: upper left panel, UR: upper right panel, LL: lower left panel, LR: lower right panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Comparison of the effect of retinoic acid (RA) treatment and nontreatment on regulatory T cells (Tregs) induction by peptide number 14 from Porphyromonas gingivalis heat shock protein 60 (HSP60). (A) T cells were stained for CD4, CD25, and Foxp3 and analyzed by fluorescence activated cell sorter. The percentage of peptide-specific Tregs induced increased from 23.70% to 32.54% (subject 1 in C), and it increased from 27.57% to 34.83% (subject 2 in C) for peptide number 14 from P. gingivalis HSP60 by the addition of RA. (B) The bar graph shows the percentage of peptide-specific Tregs among T cells in two subjects. The difference between cases with and without RA failed to reach statistical significance. UL: upper left panel, UR: upper right panel, LL: lower left panel, LR: lower right panel.
Mentions: RA has been reported to stimulate the expression of CD103 by DCs when added to the culture medium after culturing the DCs for 6 days. As shown in Fig. 4, the morphology of the DCs exhibited the typical extension of dendritic processes characteristic of maturing DCs. The CD103+ cells were increased from 4.28% to 32.67%. This characteristic phenotype expression occurred in parallel with its functional capability of inducing Tregs. After adding RA, the percentage of induced Tregs increased from 21.12% to 22.89% in subject 1, and from 27.73% to 34.46% in subject 2 for peptide number 19, respectively (Fig. 5). In case of peptide 14, it increased from 23.70% to 32.54% in subject 1 and from 27.57% to 34.83% in subject 2 (Fig. 6). There were no statistical differences in Tregs stimulated by DCs and RA-stimulated DCs for both peptide 19 and 14.

Bottom Line: Exogenous interleukin 2 was added as a coculture supplement.RA enhanced the conversion of CD103(+) DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance.RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, Pusan University School of Dentistry, Yangsan, Korea.

ABSTRACT

Purpose: Regulatory T cells (Tregs), expressing CD4 and CD25 as well as Foxp3, are known to play a pivotal role in immunoregulatory function in autoimmune diseases, cancers, and graft rejection. Dendritic cells (DCs) are considered the major antigen-presenting cells (APCs) for initiating these T-cell immune responses, of which CD103(+) DCs are derived from precursor human peripheral blood mononuclear cells (PBMCs). The aim of the present study was to evaluate the capacity of these PBMC-derived CD103(+) DCs to promote the differentiation of antigen-specific Tregs.

Methods: Monocyte-derived DCs were induced from CD14(+) monocytes from the PBMCs of 10 healthy subjects. Once the CD103(+) DCs were purified, the cell population was enriched by adding retinoic acid (RA). Peptide numbers 14 and 19 of Porphyromonas gingivalis heat shock protein 60 (HSP60) were synthesized to pulse CD103(+) DCs as a tool for presenting the peptide antigens to stimulate CD3(+) T cells that were isolated from human PBMC. Exogenous interleukin 2 was added as a coculture supplement. The antigen-specific T-cell lines established were phenotypically identified for their expression of CD4, CD25, or Foxp3.

Results: When PBMCs were used as APCs, they demonstrated only a marginal capacity to stimulate peptide-specific Tregs, whereas CD103(+) DCs showed a potent antigen presenting capability to promote the peptide-specific Tregs, especially for peptide 14. RA enhanced the conversion of CD103(+) DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance.

Conclusions: We demonstrated that CD103(+) DCs can promote antigen-specific Tregs from naive T cells, when used as APCs for an epitope peptide from P. gingivalis HSP60. RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

No MeSH data available.


Related in: MedlinePlus