Limits...
Jagged-1 is required for the expansion of CD4+ CD25+ FoxP3+ regulatory T cells and tolerogenic dendritic cells by murine mesenchymal stromal cells.

Cahill EF, Tobin LM, Carty F, Mahon BP, English K - Stem Cell Res Ther (2015)

Bottom Line: MSC, but not Jagged-1 knock down MSC, reduced pathology in a mouse model of allergic airway inflammation.Protection mediated by MSC was associated with enhanced Treg in the lung and significantly increased production of interleukin (IL)-10 in splenocytes re-stimulated with allergen.The current study suggests that MSC-mediated immune modulation involves the education and expansion of regulatory immune cells in a Jagged-1 dependent manner and provides the first report of the importance of Jagged-1 signalling in MSC protection against inflammation in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Institute of Immunology, Maynooth University, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland. emerfc@gmail.com.

ABSTRACT

Introduction: Mesenchymal stromal cells (MSC) have well defined immunomodulatory properties including the suppression of lymphocyte proliferation and inhibition of dendritic cell (DC) maturation involving both cell contact and soluble factors. These properties have made MSC attractive candidates for cellular therapy. However, the mechanism underlying these characteristics remains unclear. This study sought to investigate the mechanisms by which MSC induce a regulatory environment.

Method: Allogeneic bone marrow mesenchymal stromal cells were cultured with T cells or dendritic cells in the presence or absence of gamma secretase inhibitor to block Notch receptor signalling. T cells and dendritic cells were examined by flow cytometry for changes in phenotype marker expression. Stable knock down MSC were generated to examine the influence of Jagged 1 signalling by MSC. Both wildtype and knockdown MSC were subsequently used in vivo in an animal model of allergic airway inflammation.

Results: The Notch ligand Jagged-1 was demonstrated to be involved in MSC expansion of regulatory T cells (Treg). Additionally, MSC-induced a functional semi-mature DC phenotype, which further required Notch signalling for the expansion of Treg. MSC, but not Jagged-1 knock down MSC, reduced pathology in a mouse model of allergic airway inflammation. Protection mediated by MSC was associated with enhanced Treg in the lung and significantly increased production of interleukin (IL)-10 in splenocytes re-stimulated with allergen. Significantly less Treg and IL-10 was observed in mice treated with Jagged-1 knock down MSC.

Conclusions: The current study suggests that MSC-mediated immune modulation involves the education and expansion of regulatory immune cells in a Jagged-1 dependent manner and provides the first report of the importance of Jagged-1 signalling in MSC protection against inflammation in vivo.

No MeSH data available.


Related in: MedlinePlus

Notch is required for the induction of functionally tolerogenic DC by MSC. A two-stage proliferation assay was carried out. Stage one: BALB/c DC (0.5 × 106/ml) were pulsed with OVA (20 μg/ml) in the presence or absence of allogeneic MSC (1.5 × 105/ml), GSI (1 μM) or DMSO (vehicle control) for 24 hours. Stage two: DC were harvested and placed into a second proliferation assay with DO11.10 CD4+ T cells (1:4, 1 × 105 DC: 4 × 105/ml T cells for 72 hours. Proliferation was analysed using thymidine (5 μCi/ml) incorporation quantified as mean counts per minute (± SEM) by liquid scintillation. Data represent three studies (*, P < 0.05; **, P < 0.01). DC, dendritic cells; DMSO, dimethyl sulfoxide; GSI, gamma secretase inhibitor; MSC, mesenchymal stem cells; OVA, ovalbumin; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4414370&req=5

Fig2: Notch is required for the induction of functionally tolerogenic DC by MSC. A two-stage proliferation assay was carried out. Stage one: BALB/c DC (0.5 × 106/ml) were pulsed with OVA (20 μg/ml) in the presence or absence of allogeneic MSC (1.5 × 105/ml), GSI (1 μM) or DMSO (vehicle control) for 24 hours. Stage two: DC were harvested and placed into a second proliferation assay with DO11.10 CD4+ T cells (1:4, 1 × 105 DC: 4 × 105/ml T cells for 72 hours. Proliferation was analysed using thymidine (5 μCi/ml) incorporation quantified as mean counts per minute (± SEM) by liquid scintillation. Data represent three studies (*, P < 0.05; **, P < 0.01). DC, dendritic cells; DMSO, dimethyl sulfoxide; GSI, gamma secretase inhibitor; MSC, mesenchymal stem cells; OVA, ovalbumin; SEM, standard error of the mean.

Mentions: Murine MSC suppress the maturation, migration and antigen presentation of DC in vitro ([21] and Additional file 4: Figure S4A,B) leading to the generation of a tDC population. Although wild-type MSC prevented DC antigen presentation, knocking down Jagged-1 in MSC partially reversed this effect (Additional file 4: Figure S4C). The importance of Notch signalling in allogeneic MSC induction of tDC was investigated using a co-culture system. BALB/c DC were pulsed with OVA in the presence or absence of C57BL/6 MSC. DC were then harvested by gentle aspiration from adherent MSC and cultured with OVA specific DO11.10 CD4+ T cells. DC pulsed with OVA that had not encountered MSC, were capable of supporting DO11.10 CD4+ T cell proliferation (Figure 2). In contrast, DC pulsed with OVA in the presence of MSC were significantly less proficient in supporting CD4+ T cell proliferation (Figure 2) indicating that MSC encounter impairs DC capacity to promote T cell proliferation.Figure 2


Jagged-1 is required for the expansion of CD4+ CD25+ FoxP3+ regulatory T cells and tolerogenic dendritic cells by murine mesenchymal stromal cells.

Cahill EF, Tobin LM, Carty F, Mahon BP, English K - Stem Cell Res Ther (2015)

Notch is required for the induction of functionally tolerogenic DC by MSC. A two-stage proliferation assay was carried out. Stage one: BALB/c DC (0.5 × 106/ml) were pulsed with OVA (20 μg/ml) in the presence or absence of allogeneic MSC (1.5 × 105/ml), GSI (1 μM) or DMSO (vehicle control) for 24 hours. Stage two: DC were harvested and placed into a second proliferation assay with DO11.10 CD4+ T cells (1:4, 1 × 105 DC: 4 × 105/ml T cells for 72 hours. Proliferation was analysed using thymidine (5 μCi/ml) incorporation quantified as mean counts per minute (± SEM) by liquid scintillation. Data represent three studies (*, P < 0.05; **, P < 0.01). DC, dendritic cells; DMSO, dimethyl sulfoxide; GSI, gamma secretase inhibitor; MSC, mesenchymal stem cells; OVA, ovalbumin; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4414370&req=5

Fig2: Notch is required for the induction of functionally tolerogenic DC by MSC. A two-stage proliferation assay was carried out. Stage one: BALB/c DC (0.5 × 106/ml) were pulsed with OVA (20 μg/ml) in the presence or absence of allogeneic MSC (1.5 × 105/ml), GSI (1 μM) or DMSO (vehicle control) for 24 hours. Stage two: DC were harvested and placed into a second proliferation assay with DO11.10 CD4+ T cells (1:4, 1 × 105 DC: 4 × 105/ml T cells for 72 hours. Proliferation was analysed using thymidine (5 μCi/ml) incorporation quantified as mean counts per minute (± SEM) by liquid scintillation. Data represent three studies (*, P < 0.05; **, P < 0.01). DC, dendritic cells; DMSO, dimethyl sulfoxide; GSI, gamma secretase inhibitor; MSC, mesenchymal stem cells; OVA, ovalbumin; SEM, standard error of the mean.
Mentions: Murine MSC suppress the maturation, migration and antigen presentation of DC in vitro ([21] and Additional file 4: Figure S4A,B) leading to the generation of a tDC population. Although wild-type MSC prevented DC antigen presentation, knocking down Jagged-1 in MSC partially reversed this effect (Additional file 4: Figure S4C). The importance of Notch signalling in allogeneic MSC induction of tDC was investigated using a co-culture system. BALB/c DC were pulsed with OVA in the presence or absence of C57BL/6 MSC. DC were then harvested by gentle aspiration from adherent MSC and cultured with OVA specific DO11.10 CD4+ T cells. DC pulsed with OVA that had not encountered MSC, were capable of supporting DO11.10 CD4+ T cell proliferation (Figure 2). In contrast, DC pulsed with OVA in the presence of MSC were significantly less proficient in supporting CD4+ T cell proliferation (Figure 2) indicating that MSC encounter impairs DC capacity to promote T cell proliferation.Figure 2

Bottom Line: MSC, but not Jagged-1 knock down MSC, reduced pathology in a mouse model of allergic airway inflammation.Protection mediated by MSC was associated with enhanced Treg in the lung and significantly increased production of interleukin (IL)-10 in splenocytes re-stimulated with allergen.The current study suggests that MSC-mediated immune modulation involves the education and expansion of regulatory immune cells in a Jagged-1 dependent manner and provides the first report of the importance of Jagged-1 signalling in MSC protection against inflammation in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Institute of Immunology, Maynooth University, National University of Ireland Maynooth, Maynooth, Co. Kildare, Ireland. emerfc@gmail.com.

ABSTRACT

Introduction: Mesenchymal stromal cells (MSC) have well defined immunomodulatory properties including the suppression of lymphocyte proliferation and inhibition of dendritic cell (DC) maturation involving both cell contact and soluble factors. These properties have made MSC attractive candidates for cellular therapy. However, the mechanism underlying these characteristics remains unclear. This study sought to investigate the mechanisms by which MSC induce a regulatory environment.

Method: Allogeneic bone marrow mesenchymal stromal cells were cultured with T cells or dendritic cells in the presence or absence of gamma secretase inhibitor to block Notch receptor signalling. T cells and dendritic cells were examined by flow cytometry for changes in phenotype marker expression. Stable knock down MSC were generated to examine the influence of Jagged 1 signalling by MSC. Both wildtype and knockdown MSC were subsequently used in vivo in an animal model of allergic airway inflammation.

Results: The Notch ligand Jagged-1 was demonstrated to be involved in MSC expansion of regulatory T cells (Treg). Additionally, MSC-induced a functional semi-mature DC phenotype, which further required Notch signalling for the expansion of Treg. MSC, but not Jagged-1 knock down MSC, reduced pathology in a mouse model of allergic airway inflammation. Protection mediated by MSC was associated with enhanced Treg in the lung and significantly increased production of interleukin (IL)-10 in splenocytes re-stimulated with allergen. Significantly less Treg and IL-10 was observed in mice treated with Jagged-1 knock down MSC.

Conclusions: The current study suggests that MSC-mediated immune modulation involves the education and expansion of regulatory immune cells in a Jagged-1 dependent manner and provides the first report of the importance of Jagged-1 signalling in MSC protection against inflammation in vivo.

No MeSH data available.


Related in: MedlinePlus