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Mouse aorta-derived mesenchymal progenitor cells contribute to and enhance the immune response of macrophage cells under inflammatory conditions.

Evans JF, Salvador V, George S, Trevino-Gutierrez C, Nunez C - Stem Cell Res Ther (2015)

Bottom Line: The resident mesenchymal progenitor cell is a potential contributor to vascular inflammation when in contact with inflamed and lipid-laden MΦ cells.This interaction represents an additional target in vascular disease treatment.The potential for resident cells to contribute to the local immune response should be considered when designing therapeutics targeting inflammatory vascular disease.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Research Core, Winthrop University Hospital, 222 Station Plaza North, Mineola, NY, 11501, USA. jevans@winthrop.org.

ABSTRACT

Introduction: Mesenchymal progenitor cells interact with immune cells and modulate inflammatory responses. The cellular characteristics required for this modulation are under fervent investigation. Upon interaction with macrophage cells, they can contribute to or suppress an inflammatory response. Current studies have focused on mesenchymal progenitors derived from bone marrow, adipose, and placenta. However, the arterial wall contains many mesenchymal progenitor cells, which during vascular disease progression have the potential to interact with macrophage cells. To examine the consequence of vascular-tissue progenitor cell-macrophage cell interactions in an inflammatory environment, we used a recently established mesenchymal progenitor cell line derived from the mouse aorta.

Methods: Mouse bone marrow-derived macrophage (MΦ) cells and mouse aorta-derived mesenchymal progenitor (mAo) cells were cultured alone or co-cultured directly and indirectly. Cells were treated with oxidized low-density lipoprotein (ox-LDL) or exposed to the inflammatory mediators lipopolysaccharide (LPS) and interferon-gamma (IFNγ) or both. A Toll-like receptor-4 (TLR4)-deficient macrophage cell line was used to determine the role of the mAo cells. To monitor inflammation, nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFα) secretions were measured.

Results: Mesenchymal progenitor cells isolated from aorta and cloned by high proliferative capacity (mAo) can differentiate into multiple mesenchymal lineages and are positive for several commonly used mouse mesenchymal stem cell markers (that is, CD29, CD44, CD105, CD106, and Sca-1) but are negative for CD73 and ecto-5'-nucleotidase. In co-culture with MΦ cells, they increase MΦ oxidized-LDL uptake by 52.2%. In an inflammatory environment, they synergistically and additively contribute to local production of both NO and IL-6. After exposure to ox-LDL, the inflammatory response of MΦ cells to LPS and LPS/IFNγ is muted. However, when lipid-laden MΦ cells are co-cultured with mAo cell progenitors, the muted response is recovered and the contribution by the mAo cell progenitor is dependent upon cell contact.

Conclusions: The resident mesenchymal progenitor cell is a potential contributor to vascular inflammation when in contact with inflamed and lipid-laden MΦ cells. This interaction represents an additional target in vascular disease treatment. The potential for resident cells to contribute to the local immune response should be considered when designing therapeutics targeting inflammatory vascular disease.

No MeSH data available.


Related in: MedlinePlus

Cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures. MΦ and mAo MSCs were grown separated by a 0.4-μm filter (A) and activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) for 24 hours. Supernatant of the mAo cells grown in the well and supernatant from the MΦ cells grown in the transwell were analyzed for NO content separately (B). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from both transwell and well compartments. IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; NO, nitric oxide.
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Fig2: Cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures. MΦ and mAo MSCs were grown separated by a 0.4-μm filter (A) and activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) for 24 hours. Supernatant of the mAo cells grown in the well and supernatant from the MΦ cells grown in the transwell were analyzed for NO content separately (B). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from both transwell and well compartments. IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; NO, nitric oxide.

Mentions: To determine whether cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures after activation, we used a transwell culture system (Figure 2A). mAo and MΦ cell populations were grown separated by a 0.4-μM filter which allows the passage of soluble factors between cell types but does not allow cell-cell contact. Both cell types were exposed to LPS or LPS and IFNγ, and the supernatants collected from the transwell compartment and the well compartment were analyzed for NO content separately with the expectation that NO concentrations in the compartments would be equivalent. Our results, however, reflect a gradient in NO diffusion, with NO concentrations greater near the MΦ cells in the transwell compartment. Regardless of the gradient, co-cultures grown in direct contact produce significantly greater NO than found in either compartment of the transwell culture system (Figure 2B).Figure 2


Mouse aorta-derived mesenchymal progenitor cells contribute to and enhance the immune response of macrophage cells under inflammatory conditions.

Evans JF, Salvador V, George S, Trevino-Gutierrez C, Nunez C - Stem Cell Res Ther (2015)

Cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures. MΦ and mAo MSCs were grown separated by a 0.4-μm filter (A) and activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) for 24 hours. Supernatant of the mAo cells grown in the well and supernatant from the MΦ cells grown in the transwell were analyzed for NO content separately (B). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from both transwell and well compartments. IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; NO, nitric oxide.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4414009&req=5

Fig2: Cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures. MΦ and mAo MSCs were grown separated by a 0.4-μm filter (A) and activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) for 24 hours. Supernatant of the mAo cells grown in the well and supernatant from the MΦ cells grown in the transwell were analyzed for NO content separately (B). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from both transwell and well compartments. IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; NO, nitric oxide.
Mentions: To determine whether cell-cell contact is required for the production of NO in MΦ/mAo cell co-cultures after activation, we used a transwell culture system (Figure 2A). mAo and MΦ cell populations were grown separated by a 0.4-μM filter which allows the passage of soluble factors between cell types but does not allow cell-cell contact. Both cell types were exposed to LPS or LPS and IFNγ, and the supernatants collected from the transwell compartment and the well compartment were analyzed for NO content separately with the expectation that NO concentrations in the compartments would be equivalent. Our results, however, reflect a gradient in NO diffusion, with NO concentrations greater near the MΦ cells in the transwell compartment. Regardless of the gradient, co-cultures grown in direct contact produce significantly greater NO than found in either compartment of the transwell culture system (Figure 2B).Figure 2

Bottom Line: The resident mesenchymal progenitor cell is a potential contributor to vascular inflammation when in contact with inflamed and lipid-laden MΦ cells.This interaction represents an additional target in vascular disease treatment.The potential for resident cells to contribute to the local immune response should be considered when designing therapeutics targeting inflammatory vascular disease.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Research Core, Winthrop University Hospital, 222 Station Plaza North, Mineola, NY, 11501, USA. jevans@winthrop.org.

ABSTRACT

Introduction: Mesenchymal progenitor cells interact with immune cells and modulate inflammatory responses. The cellular characteristics required for this modulation are under fervent investigation. Upon interaction with macrophage cells, they can contribute to or suppress an inflammatory response. Current studies have focused on mesenchymal progenitors derived from bone marrow, adipose, and placenta. However, the arterial wall contains many mesenchymal progenitor cells, which during vascular disease progression have the potential to interact with macrophage cells. To examine the consequence of vascular-tissue progenitor cell-macrophage cell interactions in an inflammatory environment, we used a recently established mesenchymal progenitor cell line derived from the mouse aorta.

Methods: Mouse bone marrow-derived macrophage (MΦ) cells and mouse aorta-derived mesenchymal progenitor (mAo) cells were cultured alone or co-cultured directly and indirectly. Cells were treated with oxidized low-density lipoprotein (ox-LDL) or exposed to the inflammatory mediators lipopolysaccharide (LPS) and interferon-gamma (IFNγ) or both. A Toll-like receptor-4 (TLR4)-deficient macrophage cell line was used to determine the role of the mAo cells. To monitor inflammation, nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFα) secretions were measured.

Results: Mesenchymal progenitor cells isolated from aorta and cloned by high proliferative capacity (mAo) can differentiate into multiple mesenchymal lineages and are positive for several commonly used mouse mesenchymal stem cell markers (that is, CD29, CD44, CD105, CD106, and Sca-1) but are negative for CD73 and ecto-5'-nucleotidase. In co-culture with MΦ cells, they increase MΦ oxidized-LDL uptake by 52.2%. In an inflammatory environment, they synergistically and additively contribute to local production of both NO and IL-6. After exposure to ox-LDL, the inflammatory response of MΦ cells to LPS and LPS/IFNγ is muted. However, when lipid-laden MΦ cells are co-cultured with mAo cell progenitors, the muted response is recovered and the contribution by the mAo cell progenitor is dependent upon cell contact.

Conclusions: The resident mesenchymal progenitor cell is a potential contributor to vascular inflammation when in contact with inflamed and lipid-laden MΦ cells. This interaction represents an additional target in vascular disease treatment. The potential for resident cells to contribute to the local immune response should be considered when designing therapeutics targeting inflammatory vascular disease.

No MeSH data available.


Related in: MedlinePlus