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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

With exposure to ox-LDL, MΦ and mAo cell interaction restores significant LPS- and LPS/IFNγ-induced TNFα production by MΦ cells. Secreted TNFα measured in culture supernatants of MΦ cells (A) and TLR4-MΦ cells (B) alone and in co-culture with mAo cells after being treated with or without ox-LDL (50 μg/mL) for 24 hours and then activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) is shown. MΦ cells after being exposed to ox-LDL (50 μg/mL) for 24 hours were also cultured in transwells with mAo cells present in the well as depicted in (C). Both MΦ cells in the transwell and mAo cells in the well were activated with LPS or LPS + IFNγ, and supernatants from the transwell and well were assayed for TNFα content separately (C). Ox-LDL-treated mAo and MΦ cells were exposed to CM (collected after ox-LDL exposure and LPS and LPS + IFNγ (L/I) activation) from the opposing cell type. TNFα levels in the CM were subtracted from the supernatant values, and the net production is presented in (D). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from MΦ or TLR4-MΦ cells alone under same conditions; #significantly different from non-ox-LDL-treated counterpart; †significantly different from well and transwell supernatant; ‡significantly different from cultures exposed to CM. CM, conditioned medium; IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; ox-LDL, oxidized low-density lipoprotein; TLR4-MΦ, Toll-like receptor-4-deficient macrophage; TNFα, tumor necrosis factor-alpha.
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Fig7: With exposure to ox-LDL, MΦ and mAo cell interaction restores significant LPS- and LPS/IFNγ-induced TNFα production by MΦ cells. Secreted TNFα measured in culture supernatants of MΦ cells (A) and TLR4-MΦ cells (B) alone and in co-culture with mAo cells after being treated with or without ox-LDL (50 μg/mL) for 24 hours and then activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) is shown. MΦ cells after being exposed to ox-LDL (50 μg/mL) for 24 hours were also cultured in transwells with mAo cells present in the well as depicted in (C). Both MΦ cells in the transwell and mAo cells in the well were activated with LPS or LPS + IFNγ, and supernatants from the transwell and well were assayed for TNFα content separately (C). Ox-LDL-treated mAo and MΦ cells were exposed to CM (collected after ox-LDL exposure and LPS and LPS + IFNγ (L/I) activation) from the opposing cell type. TNFα levels in the CM were subtracted from the supernatant values, and the net production is presented in (D). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from MΦ or TLR4-MΦ cells alone under same conditions; #significantly different from non-ox-LDL-treated counterpart; †significantly different from well and transwell supernatant; ‡significantly different from cultures exposed to CM. CM, conditioned medium; IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; ox-LDL, oxidized low-density lipoprotein; TLR4-MΦ, Toll-like receptor-4-deficient macrophage; TNFα, tumor necrosis factor-alpha.

Mentions: We also confirmed the cytokine Proteome Profiler results for TNFα and determined the contribution of the mAo cells to the restoration of local TNFα secretion in ox-LDL-exposed cultures by using ELISA. MSCs have been reported to be unable to produce TNFα [28,29], and we confirmed these findings in the mAo cell cultures. The TNFα protein was undetectable in the supernatant of these cells, and transcript expression was also out of detection range in mAo cells with or without treatment with inflammatory mediators (Additional file 4). After uptake of ox-LDL, LPS- and LPS/IFNγ-induced TNFα production is significantly suppressed in MΦ cell cultures by 155- and 4.6-fold, respectively. Under the same conditions, interaction of MΦ and mAo cells returns TNFα to 21.7% and 63% of levels observed in non-ox-LDL-treated MΦ/mAo cell co-cultures after treatment with LPS or LPS/IFNγ, respectively (Figure 7A).Figure 7


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)

With exposure to ox-LDL, MΦ and mAo cell interaction restores significant LPS- and LPS/IFNγ-induced TNFα production by MΦ cells. Secreted TNFα measured in culture supernatants of MΦ cells (A) and TLR4-MΦ cells (B) alone and in co-culture with mAo cells after being treated with or without ox-LDL (50 μg/mL) for 24 hours and then activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) is shown. MΦ cells after being exposed to ox-LDL (50 μg/mL) for 24 hours were also cultured in transwells with mAo cells present in the well as depicted in (C). Both MΦ cells in the transwell and mAo cells in the well were activated with LPS or LPS + IFNγ, and supernatants from the transwell and well were assayed for TNFα content separately (C). Ox-LDL-treated mAo and MΦ cells were exposed to CM (collected after ox-LDL exposure and LPS and LPS + IFNγ (L/I) activation) from the opposing cell type. TNFα levels in the CM were subtracted from the supernatant values, and the net production is presented in (D). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from MΦ or TLR4-MΦ cells alone under same conditions; #significantly different from non-ox-LDL-treated counterpart; †significantly different from well and transwell supernatant; ‡significantly different from cultures exposed to CM. CM, conditioned medium; IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; ox-LDL, oxidized low-density lipoprotein; TLR4-MΦ, Toll-like receptor-4-deficient macrophage; TNFα, tumor necrosis factor-alpha.
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Related In: Results  -  Collection

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Fig7: With exposure to ox-LDL, MΦ and mAo cell interaction restores significant LPS- and LPS/IFNγ-induced TNFα production by MΦ cells. Secreted TNFα measured in culture supernatants of MΦ cells (A) and TLR4-MΦ cells (B) alone and in co-culture with mAo cells after being treated with or without ox-LDL (50 μg/mL) for 24 hours and then activated with LPS (100 ng/mL) or with LPS (100 ng/mL) + IFNγ (250 ng/mL) is shown. MΦ cells after being exposed to ox-LDL (50 μg/mL) for 24 hours were also cultured in transwells with mAo cells present in the well as depicted in (C). Both MΦ cells in the transwell and mAo cells in the well were activated with LPS or LPS + IFNγ, and supernatants from the transwell and well were assayed for TNFα content separately (C). Ox-LDL-treated mAo and MΦ cells were exposed to CM (collected after ox-LDL exposure and LPS and LPS + IFNγ (L/I) activation) from the opposing cell type. TNFα levels in the CM were subtracted from the supernatant values, and the net production is presented in (D). Data are presented as mean ± standard error of the mean and are representative of three experiments each with n = 4. *Significantly different from MΦ or TLR4-MΦ cells alone under same conditions; #significantly different from non-ox-LDL-treated counterpart; †significantly different from well and transwell supernatant; ‡significantly different from cultures exposed to CM. CM, conditioned medium; IFNγ, interferon-gamma; LPS, lipopolysaccharide; MΦ, bone marrow-derived macrophage; mAo, mouse aorta-derived mesenchymal progenitor; MSC, mesenchymal stem cell; ox-LDL, oxidized low-density lipoprotein; TLR4-MΦ, Toll-like receptor-4-deficient macrophage; TNFα, tumor necrosis factor-alpha.
Mentions: We also confirmed the cytokine Proteome Profiler results for TNFα and determined the contribution of the mAo cells to the restoration of local TNFα secretion in ox-LDL-exposed cultures by using ELISA. MSCs have been reported to be unable to produce TNFα [28,29], and we confirmed these findings in the mAo cell cultures. The TNFα protein was undetectable in the supernatant of these cells, and transcript expression was also out of detection range in mAo cells with or without treatment with inflammatory mediators (Additional file 4). After uptake of ox-LDL, LPS- and LPS/IFNγ-induced TNFα production is significantly suppressed in MΦ cell cultures by 155- and 4.6-fold, respectively. Under the same conditions, interaction of MΦ and mAo cells returns TNFα to 21.7% and 63% of levels observed in non-ox-LDL-treated MΦ/mAo cell co-cultures after treatment with LPS or LPS/IFNγ, respectively (Figure 7A).Figure 7

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