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Bone marrow-derived progenitor cells in end-stage lung disease patients.

Gilpin SE, Lung K, de Couto GT, Cypel M, Sato M, Singer LG, Keshavjee S, Waddell TK - BMC Pulm Med (2013)

Bottom Line: An increase in CD45⁺Collagen-1⁺ fibrocytes was found in pulmonary fibrosis and bronchiolitis obliterans patients.Plasma cytokine levels differed between disease groups, with a significant correlation between SCGF-β and CCSP⁺ cells and between Monocyte Chemotactic Protein-1 and fibrocytes.An increase in the novel CCSP⁺ epithelial-like progenitors in cystic fibrosis patients was found.

View Article: PubMed Central - HTML - PubMed

Affiliation: Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, Toronto General Hospital, University Health Network, University of Toronto, North Wing, 9N - 949, 200 Elizabeth Street, Toronto, ON M5G 2C4, Canada.

ABSTRACT

Background: Chronic lung diseases are marked by progressive inflammation, tissue damage and remodelling. Bone marrow-derived progenitor cells may contribute to these processes. The objectives of this study were to (1) to quantify CD45⁺Collagen-1⁺ fibrocytes and a novel epithelial-like population of bone marrow-derived cells, which express Clara Cell Secretory Protein, in patients at the time of lung transplant and (2) to evaluate mediators that may act to recruit these cells during injury.

Methods: Using an observational design, progenitor cells were quantified by flow cytometry from both bone marrow (BM) and peripheral blood (PB). Migration was tested using in vitro transwell assays. Multiplex bead-based assays were used to quantify plasma cytokines.

Results: An increase in CD45⁺Collagen-1⁺ fibrocytes was found in pulmonary fibrosis and bronchiolitis obliterans patients. Cystic fibrosis patients had an increase in CCSP⁺ cells in both the BM and PB. The proportion of CCSP⁺ cells in the BM and PB was correlated. CCSP+ cells express the chemokine receptors CCR2, CCR4, CXCR3, and CXCR4, and significantly migrated in vitro toward Stromal Derived Factor-1 (SDF-1) and Stem Cell Growth Factor-β (SCGF-β). Plasma cytokine levels differed between disease groups, with a significant correlation between SCGF-β and CCSP⁺ cells and between Monocyte Chemotactic Protein-1 and fibrocytes.

Conclusions: Different bone marrow-derived cells are found in various lung diseases. Increased fibrocytes were associated with fibrotic lung diseases. An increase in the novel CCSP⁺ epithelial-like progenitors in cystic fibrosis patients was found. These differences may be mediated by alterations in plasma cytokines responsible for cell recruitment.

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In vitro migration assay. Migration of freshly isolated peripheral blood mononuclear cells (PBMCs) or bone marrow cells (BMCs) in response to chemotactic stimuli Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES) (n = 9 control, 11 recipient PBMC, 7 BMC), Interferon gamma-induced protein 10 (IP-10) (n = 9 control, 11 recipient PBMC, 7 BMC), Stromal Derived Factor-1 (SDF-1) (n = 9 control, 13 recipient PBMC, 9 BMC), or Stem Cell Growth Factor-beta (SCGF-β) (n = 4 control, 4 recipient PBMC, 4 BMC), compared to untreated cells. Migrated cells were analyzed for CCSP+ expression and normalized to total CCSP+ cells in the starting sample. Kruskal-Wallis test with Dunn’s multiple comparison post-hoc analysis, with significance tested against untreated samples (* = p < 0.05, ** = p < .01). Boxes show the median, 25th and 75th percentiles. Whiskers represent the 2.5 and 97.5 percentiles.
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Figure 6: In vitro migration assay. Migration of freshly isolated peripheral blood mononuclear cells (PBMCs) or bone marrow cells (BMCs) in response to chemotactic stimuli Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES) (n = 9 control, 11 recipient PBMC, 7 BMC), Interferon gamma-induced protein 10 (IP-10) (n = 9 control, 11 recipient PBMC, 7 BMC), Stromal Derived Factor-1 (SDF-1) (n = 9 control, 13 recipient PBMC, 9 BMC), or Stem Cell Growth Factor-beta (SCGF-β) (n = 4 control, 4 recipient PBMC, 4 BMC), compared to untreated cells. Migrated cells were analyzed for CCSP+ expression and normalized to total CCSP+ cells in the starting sample. Kruskal-Wallis test with Dunn’s multiple comparison post-hoc analysis, with significance tested against untreated samples (* = p < 0.05, ** = p < .01). Boxes show the median, 25th and 75th percentiles. Whiskers represent the 2.5 and 97.5 percentiles.

Mentions: To further investigate the ability of CCSP+ cells to migrate in response to chemotactic mediators, in vitro transwell assays were utilized. Migration of bone marrow or peripheral blood cells (BMCs) freshly isolated from end-stage lung disease patients was investigated in response to the chemotactic stimuli RANTES, IP-10, SDF-1, or SCGF-β and compared to untreated cells (Figure 6). A significant migratory response of CCSP+ cells toward SDF-1 was identified for CCSP+ PBMCs from control and lung recipient samples, as well as from BMCs from lung recipients, compared to untreated cells in the absence of any chemotactic stimuli. In addition, significant migration in response to SCGF-β was also found for CCSP+ BMCs and PBMCs isolated from end-stage lung disease patients (p < 0.05), while no significant migratory response was found for CCSP+ PBMCs isolated from healthy controls (Figure 6).


Bone marrow-derived progenitor cells in end-stage lung disease patients.

Gilpin SE, Lung K, de Couto GT, Cypel M, Sato M, Singer LG, Keshavjee S, Waddell TK - BMC Pulm Med (2013)

In vitro migration assay. Migration of freshly isolated peripheral blood mononuclear cells (PBMCs) or bone marrow cells (BMCs) in response to chemotactic stimuli Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES) (n = 9 control, 11 recipient PBMC, 7 BMC), Interferon gamma-induced protein 10 (IP-10) (n = 9 control, 11 recipient PBMC, 7 BMC), Stromal Derived Factor-1 (SDF-1) (n = 9 control, 13 recipient PBMC, 9 BMC), or Stem Cell Growth Factor-beta (SCGF-β) (n = 4 control, 4 recipient PBMC, 4 BMC), compared to untreated cells. Migrated cells were analyzed for CCSP+ expression and normalized to total CCSP+ cells in the starting sample. Kruskal-Wallis test with Dunn’s multiple comparison post-hoc analysis, with significance tested against untreated samples (* = p < 0.05, ** = p < .01). Boxes show the median, 25th and 75th percentiles. Whiskers represent the 2.5 and 97.5 percentiles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750607&req=5

Figure 6: In vitro migration assay. Migration of freshly isolated peripheral blood mononuclear cells (PBMCs) or bone marrow cells (BMCs) in response to chemotactic stimuli Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES) (n = 9 control, 11 recipient PBMC, 7 BMC), Interferon gamma-induced protein 10 (IP-10) (n = 9 control, 11 recipient PBMC, 7 BMC), Stromal Derived Factor-1 (SDF-1) (n = 9 control, 13 recipient PBMC, 9 BMC), or Stem Cell Growth Factor-beta (SCGF-β) (n = 4 control, 4 recipient PBMC, 4 BMC), compared to untreated cells. Migrated cells were analyzed for CCSP+ expression and normalized to total CCSP+ cells in the starting sample. Kruskal-Wallis test with Dunn’s multiple comparison post-hoc analysis, with significance tested against untreated samples (* = p < 0.05, ** = p < .01). Boxes show the median, 25th and 75th percentiles. Whiskers represent the 2.5 and 97.5 percentiles.
Mentions: To further investigate the ability of CCSP+ cells to migrate in response to chemotactic mediators, in vitro transwell assays were utilized. Migration of bone marrow or peripheral blood cells (BMCs) freshly isolated from end-stage lung disease patients was investigated in response to the chemotactic stimuli RANTES, IP-10, SDF-1, or SCGF-β and compared to untreated cells (Figure 6). A significant migratory response of CCSP+ cells toward SDF-1 was identified for CCSP+ PBMCs from control and lung recipient samples, as well as from BMCs from lung recipients, compared to untreated cells in the absence of any chemotactic stimuli. In addition, significant migration in response to SCGF-β was also found for CCSP+ BMCs and PBMCs isolated from end-stage lung disease patients (p < 0.05), while no significant migratory response was found for CCSP+ PBMCs isolated from healthy controls (Figure 6).

Bottom Line: An increase in CD45⁺Collagen-1⁺ fibrocytes was found in pulmonary fibrosis and bronchiolitis obliterans patients.Plasma cytokine levels differed between disease groups, with a significant correlation between SCGF-β and CCSP⁺ cells and between Monocyte Chemotactic Protein-1 and fibrocytes.An increase in the novel CCSP⁺ epithelial-like progenitors in cystic fibrosis patients was found.

View Article: PubMed Central - HTML - PubMed

Affiliation: Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, Toronto General Hospital, University Health Network, University of Toronto, North Wing, 9N - 949, 200 Elizabeth Street, Toronto, ON M5G 2C4, Canada.

ABSTRACT

Background: Chronic lung diseases are marked by progressive inflammation, tissue damage and remodelling. Bone marrow-derived progenitor cells may contribute to these processes. The objectives of this study were to (1) to quantify CD45⁺Collagen-1⁺ fibrocytes and a novel epithelial-like population of bone marrow-derived cells, which express Clara Cell Secretory Protein, in patients at the time of lung transplant and (2) to evaluate mediators that may act to recruit these cells during injury.

Methods: Using an observational design, progenitor cells were quantified by flow cytometry from both bone marrow (BM) and peripheral blood (PB). Migration was tested using in vitro transwell assays. Multiplex bead-based assays were used to quantify plasma cytokines.

Results: An increase in CD45⁺Collagen-1⁺ fibrocytes was found in pulmonary fibrosis and bronchiolitis obliterans patients. Cystic fibrosis patients had an increase in CCSP⁺ cells in both the BM and PB. The proportion of CCSP⁺ cells in the BM and PB was correlated. CCSP+ cells express the chemokine receptors CCR2, CCR4, CXCR3, and CXCR4, and significantly migrated in vitro toward Stromal Derived Factor-1 (SDF-1) and Stem Cell Growth Factor-β (SCGF-β). Plasma cytokine levels differed between disease groups, with a significant correlation between SCGF-β and CCSP⁺ cells and between Monocyte Chemotactic Protein-1 and fibrocytes.

Conclusions: Different bone marrow-derived cells are found in various lung diseases. Increased fibrocytes were associated with fibrotic lung diseases. An increase in the novel CCSP⁺ epithelial-like progenitors in cystic fibrosis patients was found. These differences may be mediated by alterations in plasma cytokines responsible for cell recruitment.

Show MeSH
Related in: MedlinePlus