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Human placenta-derived adherent cells induce tolerogenic immune responses.

Liu W, Morschauser A, Zhang X, Lu X, Gleason J, He S, Chen HJ, Jankovic V, Ye Q, Labazzo K, Herzberg U, Albert VR, Abbot SE, Liang B, Hariri R - Clin Transl Immunology (2014)

Bottom Line: These tolerogenic DC failed to induce allogeneic T-cell proliferation and differentiation toward Th1, but skewed T-cell differentiation toward Th2.Inhibition of cyclo-oxygenase-2 activity resulted in a significant, but not complete, abrogation of PDAC cells' effects on DC phenotype and function, implying a role for prostaglandin E2 in PDAC-mediated immunomodulation.This study identifies modulation of DC differentiation toward immune tolerance as a key mechanism underlying the immunomodulatory activities of PDAC cells.

View Article: PubMed Central - PubMed

Affiliation: Celgene Cellular Therapeutics , Warren, NJ, USA.

ABSTRACT
Human placenta-derived adherent cells (PDAC cells) are a culture expanded, undifferentiated mesenchymal-like population derived from full-term placental tissue, with immunomodulatory and anti-inflammatory properties. PDA-001 (cenplacel-L), an intravenous formulation of PDAC cells, is in clinical development for the treatment of autoimmune and inflammatory diseases. To elucidate the mechanisms underlying the immunoregulatory properties of PDAC cells, we investigated their effects on immune cell populations, including T cells and dendritic cells (DC) in vitro and in vivo. PDAC cells suppressed T-cell proliferation in an OT-II T-cell adoptive transfer model, reduced the severity of myelin oligodendrocyte glycoprotein peptide-induced experimental autoimmune encephalomyelitis and ameliorated inflammation in a delayed type hypersensitivity response model. In vitro, PDAC cells suppressed T-cell proliferation and inhibited Th1 and Th17 differentiation. Analysis of tissues derived from PDAC cell-treated animals revealed diminished CD86 expression on splenic DC, suggesting that they can also modulate DC populations. Furthermore, PDAC cells modulate the differentiation and maturation of mouse bone marrow-derived DC. Similarly, human DC differentiated from CD14(+) monocytes in the presence of PDAC cells acquired a tolerogenic phenotype. These tolerogenic DC failed to induce allogeneic T-cell proliferation and differentiation toward Th1, but skewed T-cell differentiation toward Th2. Inhibition of cyclo-oxygenase-2 activity resulted in a significant, but not complete, abrogation of PDAC cells' effects on DC phenotype and function, implying a role for prostaglandin E2 in PDAC-mediated immunomodulation. This study identifies modulation of DC differentiation toward immune tolerance as a key mechanism underlying the immunomodulatory activities of PDAC cells.

No MeSH data available.


Related in: MedlinePlus

PDAC-CM can induce tolerogenic human DC in vitro. (a) Flow cytometry analysis of cell surface expression of CD1a, CD123, CD14, CD86hi and PD-L1 by iDC. iDC were generated from peripheral blood monocytes after 4 days in culture with GM-CSF and IL-4, in the presence or absence of various concentrations of PDAC-CM as indicated. (b, c) iDC generated in the presence or absence of various concentrations of PDAC-CM were treated with LPS plus IFN-γ and analyzed by flow cytometry for cell surface CD86 and CD83 expression, intracellular IL-12 production (b) and secretion of IL-12, IL-23 and IL-10 (c). iDC: before LPS and IFN-γ treatment; mDC: LPS and IFN-γ-stimulated iDC; PDAC-iDC (%CM): iDC generated in the presence of indicated concentration of PDAC-CM; PDAC-mDC (%CM): LPS plus IFN-γ-stimulated PDAC-iDC. Dotted lines indicate isotype-matched control antibody staining. Numbers indicate percent positive cells. Results are expressed as mean±s.e.m. of cytokine levels in 1.5 × 105 ml−1 of mDC obtained from the analysis of three independent experiments. *P<0.05, **P<0.01, compared with mDC control. (d, e) Effects of PDAC-mDC on T-cell proliferation and differentiation in MLR. iDC (before LPS plus IFN-γ treatment), mDC (LPS plus IFN-γ stimulated iDC) and PDAC-mDC (LPS plus IFN-γ stimulated PDAC-iDC differentiated with 50% of PDAC-CM) were cocultured with allogeneic CD4+ naive T cells. (d) Percent proliferation of CFSE-labeled CD4+ T cells, analyzed after 5-day culture with the DC:T at a ratio of 1:100. Dotted lines indicate CFSE-labeled T cells without DC. Numbers in top left corners indicate percent proliferated cells. (e) IFN-γ+ and IL-4+ production by CD4+ allogeneic T cells analyzed via intracellular cytokine staining after a 6-day mixed culture with a DC:T ratio of 1:20. Plots are gated on CD4+ cells and quadrant frequencies are indicated.
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fig4: PDAC-CM can induce tolerogenic human DC in vitro. (a) Flow cytometry analysis of cell surface expression of CD1a, CD123, CD14, CD86hi and PD-L1 by iDC. iDC were generated from peripheral blood monocytes after 4 days in culture with GM-CSF and IL-4, in the presence or absence of various concentrations of PDAC-CM as indicated. (b, c) iDC generated in the presence or absence of various concentrations of PDAC-CM were treated with LPS plus IFN-γ and analyzed by flow cytometry for cell surface CD86 and CD83 expression, intracellular IL-12 production (b) and secretion of IL-12, IL-23 and IL-10 (c). iDC: before LPS and IFN-γ treatment; mDC: LPS and IFN-γ-stimulated iDC; PDAC-iDC (%CM): iDC generated in the presence of indicated concentration of PDAC-CM; PDAC-mDC (%CM): LPS plus IFN-γ-stimulated PDAC-iDC. Dotted lines indicate isotype-matched control antibody staining. Numbers indicate percent positive cells. Results are expressed as mean±s.e.m. of cytokine levels in 1.5 × 105 ml−1 of mDC obtained from the analysis of three independent experiments. *P<0.05, **P<0.01, compared with mDC control. (d, e) Effects of PDAC-mDC on T-cell proliferation and differentiation in MLR. iDC (before LPS plus IFN-γ treatment), mDC (LPS plus IFN-γ stimulated iDC) and PDAC-mDC (LPS plus IFN-γ stimulated PDAC-iDC differentiated with 50% of PDAC-CM) were cocultured with allogeneic CD4+ naive T cells. (d) Percent proliferation of CFSE-labeled CD4+ T cells, analyzed after 5-day culture with the DC:T at a ratio of 1:100. Dotted lines indicate CFSE-labeled T cells without DC. Numbers in top left corners indicate percent proliferated cells. (e) IFN-γ+ and IL-4+ production by CD4+ allogeneic T cells analyzed via intracellular cytokine staining after a 6-day mixed culture with a DC:T ratio of 1:20. Plots are gated on CD4+ cells and quadrant frequencies are indicated.

Mentions: The effects of PDAC cells on DC differentiation were further examined using human DC. Immature DC (iDC) were differentiated from human peripheral CD14+ monocytes in medium containing GM-CSF and IL-4, and cultured in the presence or absence of PDAC cells or PDAC-CM. iDC were then activated to develop into mature DC (mDC) by subsequent stimulation with LPS and IFN-γ. DC differentiation, indicated by the upregulation of DC markers and costimulatory molecules and downregulation of monocyte markers, was inhibited in the presence of PDAC cells. PDAC cells (Supplementary Online Figure 5a) or PDAC-CM (Figure 4a) reduced conventional DC (cDC) marker CD1a expression and enhanced expression of the plasmacytoid DC (pDC) marker CD123 and the monocyte lineage marker CD14 on iDC in a dose-dependent manner. DC differentiated in the presence of PDAC cells (PDAC-iDC) (Supplementary Online Figure 5a) or PDAC-CM also demonstrated dose-dependent reduction of costimulatory molecule CD86 expression and enhancement of PD-L1 expression (Figure 4a). Mature DC that were originally differentiated in the presence of PDAC cells or PDAC-CM (PDAC-mDC) were further impaired in their ability to express CD86 and induce the human DC maturation marker CD83, in response to LPS and IFN-γ stimulation (Supplementary Online Figure 5b and Figure 4b). Moreover, intracellular staining revealed reduced production of the Th1-polarizing cytokine IL-12 (Supplementary Online Figure 5b and Figure 4b). Secretory IL-12 and IL-23 production was also reduced along with an increase in IL-10 production in the culture supernatant of PDAC-mDC induced from PDAC-iDC that were originally differentiated in the presence of PDAC cells (Supplementary Online Figure 5c) or PDAC-CM (Figure 4c). Taken together, these findings suggest that PDAC cells can influence human MoDC differentiation toward a tolerogenic phenotype.


Human placenta-derived adherent cells induce tolerogenic immune responses.

Liu W, Morschauser A, Zhang X, Lu X, Gleason J, He S, Chen HJ, Jankovic V, Ye Q, Labazzo K, Herzberg U, Albert VR, Abbot SE, Liang B, Hariri R - Clin Transl Immunology (2014)

PDAC-CM can induce tolerogenic human DC in vitro. (a) Flow cytometry analysis of cell surface expression of CD1a, CD123, CD14, CD86hi and PD-L1 by iDC. iDC were generated from peripheral blood monocytes after 4 days in culture with GM-CSF and IL-4, in the presence or absence of various concentrations of PDAC-CM as indicated. (b, c) iDC generated in the presence or absence of various concentrations of PDAC-CM were treated with LPS plus IFN-γ and analyzed by flow cytometry for cell surface CD86 and CD83 expression, intracellular IL-12 production (b) and secretion of IL-12, IL-23 and IL-10 (c). iDC: before LPS and IFN-γ treatment; mDC: LPS and IFN-γ-stimulated iDC; PDAC-iDC (%CM): iDC generated in the presence of indicated concentration of PDAC-CM; PDAC-mDC (%CM): LPS plus IFN-γ-stimulated PDAC-iDC. Dotted lines indicate isotype-matched control antibody staining. Numbers indicate percent positive cells. Results are expressed as mean±s.e.m. of cytokine levels in 1.5 × 105 ml−1 of mDC obtained from the analysis of three independent experiments. *P<0.05, **P<0.01, compared with mDC control. (d, e) Effects of PDAC-mDC on T-cell proliferation and differentiation in MLR. iDC (before LPS plus IFN-γ treatment), mDC (LPS plus IFN-γ stimulated iDC) and PDAC-mDC (LPS plus IFN-γ stimulated PDAC-iDC differentiated with 50% of PDAC-CM) were cocultured with allogeneic CD4+ naive T cells. (d) Percent proliferation of CFSE-labeled CD4+ T cells, analyzed after 5-day culture with the DC:T at a ratio of 1:100. Dotted lines indicate CFSE-labeled T cells without DC. Numbers in top left corners indicate percent proliferated cells. (e) IFN-γ+ and IL-4+ production by CD4+ allogeneic T cells analyzed via intracellular cytokine staining after a 6-day mixed culture with a DC:T ratio of 1:20. Plots are gated on CD4+ cells and quadrant frequencies are indicated.
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fig4: PDAC-CM can induce tolerogenic human DC in vitro. (a) Flow cytometry analysis of cell surface expression of CD1a, CD123, CD14, CD86hi and PD-L1 by iDC. iDC were generated from peripheral blood monocytes after 4 days in culture with GM-CSF and IL-4, in the presence or absence of various concentrations of PDAC-CM as indicated. (b, c) iDC generated in the presence or absence of various concentrations of PDAC-CM were treated with LPS plus IFN-γ and analyzed by flow cytometry for cell surface CD86 and CD83 expression, intracellular IL-12 production (b) and secretion of IL-12, IL-23 and IL-10 (c). iDC: before LPS and IFN-γ treatment; mDC: LPS and IFN-γ-stimulated iDC; PDAC-iDC (%CM): iDC generated in the presence of indicated concentration of PDAC-CM; PDAC-mDC (%CM): LPS plus IFN-γ-stimulated PDAC-iDC. Dotted lines indicate isotype-matched control antibody staining. Numbers indicate percent positive cells. Results are expressed as mean±s.e.m. of cytokine levels in 1.5 × 105 ml−1 of mDC obtained from the analysis of three independent experiments. *P<0.05, **P<0.01, compared with mDC control. (d, e) Effects of PDAC-mDC on T-cell proliferation and differentiation in MLR. iDC (before LPS plus IFN-γ treatment), mDC (LPS plus IFN-γ stimulated iDC) and PDAC-mDC (LPS plus IFN-γ stimulated PDAC-iDC differentiated with 50% of PDAC-CM) were cocultured with allogeneic CD4+ naive T cells. (d) Percent proliferation of CFSE-labeled CD4+ T cells, analyzed after 5-day culture with the DC:T at a ratio of 1:100. Dotted lines indicate CFSE-labeled T cells without DC. Numbers in top left corners indicate percent proliferated cells. (e) IFN-γ+ and IL-4+ production by CD4+ allogeneic T cells analyzed via intracellular cytokine staining after a 6-day mixed culture with a DC:T ratio of 1:20. Plots are gated on CD4+ cells and quadrant frequencies are indicated.
Mentions: The effects of PDAC cells on DC differentiation were further examined using human DC. Immature DC (iDC) were differentiated from human peripheral CD14+ monocytes in medium containing GM-CSF and IL-4, and cultured in the presence or absence of PDAC cells or PDAC-CM. iDC were then activated to develop into mature DC (mDC) by subsequent stimulation with LPS and IFN-γ. DC differentiation, indicated by the upregulation of DC markers and costimulatory molecules and downregulation of monocyte markers, was inhibited in the presence of PDAC cells. PDAC cells (Supplementary Online Figure 5a) or PDAC-CM (Figure 4a) reduced conventional DC (cDC) marker CD1a expression and enhanced expression of the plasmacytoid DC (pDC) marker CD123 and the monocyte lineage marker CD14 on iDC in a dose-dependent manner. DC differentiated in the presence of PDAC cells (PDAC-iDC) (Supplementary Online Figure 5a) or PDAC-CM also demonstrated dose-dependent reduction of costimulatory molecule CD86 expression and enhancement of PD-L1 expression (Figure 4a). Mature DC that were originally differentiated in the presence of PDAC cells or PDAC-CM (PDAC-mDC) were further impaired in their ability to express CD86 and induce the human DC maturation marker CD83, in response to LPS and IFN-γ stimulation (Supplementary Online Figure 5b and Figure 4b). Moreover, intracellular staining revealed reduced production of the Th1-polarizing cytokine IL-12 (Supplementary Online Figure 5b and Figure 4b). Secretory IL-12 and IL-23 production was also reduced along with an increase in IL-10 production in the culture supernatant of PDAC-mDC induced from PDAC-iDC that were originally differentiated in the presence of PDAC cells (Supplementary Online Figure 5c) or PDAC-CM (Figure 4c). Taken together, these findings suggest that PDAC cells can influence human MoDC differentiation toward a tolerogenic phenotype.

Bottom Line: These tolerogenic DC failed to induce allogeneic T-cell proliferation and differentiation toward Th1, but skewed T-cell differentiation toward Th2.Inhibition of cyclo-oxygenase-2 activity resulted in a significant, but not complete, abrogation of PDAC cells' effects on DC phenotype and function, implying a role for prostaglandin E2 in PDAC-mediated immunomodulation.This study identifies modulation of DC differentiation toward immune tolerance as a key mechanism underlying the immunomodulatory activities of PDAC cells.

View Article: PubMed Central - PubMed

Affiliation: Celgene Cellular Therapeutics , Warren, NJ, USA.

ABSTRACT
Human placenta-derived adherent cells (PDAC cells) are a culture expanded, undifferentiated mesenchymal-like population derived from full-term placental tissue, with immunomodulatory and anti-inflammatory properties. PDA-001 (cenplacel-L), an intravenous formulation of PDAC cells, is in clinical development for the treatment of autoimmune and inflammatory diseases. To elucidate the mechanisms underlying the immunoregulatory properties of PDAC cells, we investigated their effects on immune cell populations, including T cells and dendritic cells (DC) in vitro and in vivo. PDAC cells suppressed T-cell proliferation in an OT-II T-cell adoptive transfer model, reduced the severity of myelin oligodendrocyte glycoprotein peptide-induced experimental autoimmune encephalomyelitis and ameliorated inflammation in a delayed type hypersensitivity response model. In vitro, PDAC cells suppressed T-cell proliferation and inhibited Th1 and Th17 differentiation. Analysis of tissues derived from PDAC cell-treated animals revealed diminished CD86 expression on splenic DC, suggesting that they can also modulate DC populations. Furthermore, PDAC cells modulate the differentiation and maturation of mouse bone marrow-derived DC. Similarly, human DC differentiated from CD14(+) monocytes in the presence of PDAC cells acquired a tolerogenic phenotype. These tolerogenic DC failed to induce allogeneic T-cell proliferation and differentiation toward Th1, but skewed T-cell differentiation toward Th2. Inhibition of cyclo-oxygenase-2 activity resulted in a significant, but not complete, abrogation of PDAC cells' effects on DC phenotype and function, implying a role for prostaglandin E2 in PDAC-mediated immunomodulation. This study identifies modulation of DC differentiation toward immune tolerance as a key mechanism underlying the immunomodulatory activities of PDAC cells.

No MeSH data available.


Related in: MedlinePlus