Limits...
Increased platelet reactivity in idiopathic pulmonary fibrosis is mediated by a plasma factor.

Crooks MG, Fahim A, Naseem KM, Morice AH, Hart SP - PLoS ONE (2014)

Bottom Line: Platelet P-selectin expression was significantly greater in IPF patients than controls following stimulation with 0.1 µM ADP (1.9% positive ±0.5 (mean ± SEM) versus 0.7%±0.1; p = 0.03), 1 µM ADP (9.8%±1.3 versus 3.3%±0.8; p<0.01) and 10 µM ADP (41.3%±4.2 versus 22.5%±2.6; p<0.01).Re-suspension of control platelets in plasma taken from subjects with IPF resulted in increased platelet activation compared to control plasma.IPF patients exhibit increased platelet reactivity compared with controls.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular and Metabolic Research, Hull York Medical School, Cottingham, United Kingdom.

ABSTRACT

Introduction: Idiopathic Pulmonary Fibrosis (IPF) is a progressive, incurable fibrotic interstitial lung disease with a prognosis worse than many cancers. Its pathogenesis is poorly understood. Activated platelets can release pro-fibrotic mediators that have the potential to contribute to lung fibrosis. We determine platelet reactivity in subjects with IPF compared to age-matched controls.

Methods: Whole blood flow cytometry was used to measure platelet-monocyte aggregate formation, platelet P-selectin expression and platelet fibrinogen binding at basal levels and following stimulation with platelet agonists. A plasma swap approach was used to assess the effect of IPF plasma on control platelets.

Results: Subjects with IPF showed greater platelet reactivity than controls. Platelet P-selectin expression was significantly greater in IPF patients than controls following stimulation with 0.1 µM ADP (1.9% positive ±0.5 (mean ± SEM) versus 0.7%±0.1; p = 0.03), 1 µM ADP (9.8%±1.3 versus 3.3%±0.8; p<0.01) and 10 µM ADP (41.3%±4.2 versus 22.5%±2.6; p<0.01). Platelet fibrinogen binding was also increased, and platelet activation resulted in increased platelet-monocyte aggregate formation in IPF patients. Re-suspension of control platelets in plasma taken from subjects with IPF resulted in increased platelet activation compared to control plasma.

Conclusions: IPF patients exhibit increased platelet reactivity compared with controls. This hyperactivity may result from the plasma environment since control platelets exhibit increased activation when exposed to IPF plasma.

Show MeSH

Related in: MedlinePlus

Platelet-monocyte aggregate formation in IPF patients and controls.A. Scatter plot demonstrating the different cell populations on whole blood flow cytometry, gated on monocytes. B. Quadrant plot demonstrating platelet-monocyte aggregates in the right upper quadrant. X-axis (FL1) shows CD42b staining and y-axis (FL2) shows CD14 staining. C. Percentage of monocytes forming aggregates with platelets at basal levels and in response to the platelet agonists ADP (0.1–10 µM) or TFLLR (1–10 µM). * P≤0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4206466&req=5

pone-0111347-g001: Platelet-monocyte aggregate formation in IPF patients and controls.A. Scatter plot demonstrating the different cell populations on whole blood flow cytometry, gated on monocytes. B. Quadrant plot demonstrating platelet-monocyte aggregates in the right upper quadrant. X-axis (FL1) shows CD42b staining and y-axis (FL2) shows CD14 staining. C. Percentage of monocytes forming aggregates with platelets at basal levels and in response to the platelet agonists ADP (0.1–10 µM) or TFLLR (1–10 µM). * P≤0.01.

Mentions: Patients with IPF and controls were matched for age and gender. When whole blood was stimulated with ADP (1–10 µM) or TFLLR (1–10 µM), subjects with IPF demonstrated significantly greater platelet-monocyte aggregate formation compared with controls (Figure 1 and table S3 in file S1). The percentage of monocytes with bound platelets was significantly greater in IPF patients following stimulation with 1 µM ADP (29.4%±4.1 (mean ± SEM) versus 16.1%±1.5 in controls; p = <0.01) and 10 µM ADP (44.8%±3.3 versus 32.1%±3.5; p = 0.01). Platelet-monocyte aggregate formation in blood from IPF patients was also significantly increased following stimulation with 5 µM and 10 µM TFLLR (figure 1) when compared to controls. Under basal (unstimulated) conditions, blood from IPF patients exhibited greater platelet-monocyte aggregate formation than controls but this did not reach statistical significance (18.2%±3.9 versus 13.7%±1.4; p = 0.3). The calcium-dependent nature of the interaction between platelets and monocytes was confirmed by addition of EDTA, which resulted in a marked reduction in aggregate formation that did not increase in response to agonist stimulation and did not vary significantly between the groups (figure 2).


Increased platelet reactivity in idiopathic pulmonary fibrosis is mediated by a plasma factor.

Crooks MG, Fahim A, Naseem KM, Morice AH, Hart SP - PLoS ONE (2014)

Platelet-monocyte aggregate formation in IPF patients and controls.A. Scatter plot demonstrating the different cell populations on whole blood flow cytometry, gated on monocytes. B. Quadrant plot demonstrating platelet-monocyte aggregates in the right upper quadrant. X-axis (FL1) shows CD42b staining and y-axis (FL2) shows CD14 staining. C. Percentage of monocytes forming aggregates with platelets at basal levels and in response to the platelet agonists ADP (0.1–10 µM) or TFLLR (1–10 µM). * P≤0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206466&req=5

pone-0111347-g001: Platelet-monocyte aggregate formation in IPF patients and controls.A. Scatter plot demonstrating the different cell populations on whole blood flow cytometry, gated on monocytes. B. Quadrant plot demonstrating platelet-monocyte aggregates in the right upper quadrant. X-axis (FL1) shows CD42b staining and y-axis (FL2) shows CD14 staining. C. Percentage of monocytes forming aggregates with platelets at basal levels and in response to the platelet agonists ADP (0.1–10 µM) or TFLLR (1–10 µM). * P≤0.01.
Mentions: Patients with IPF and controls were matched for age and gender. When whole blood was stimulated with ADP (1–10 µM) or TFLLR (1–10 µM), subjects with IPF demonstrated significantly greater platelet-monocyte aggregate formation compared with controls (Figure 1 and table S3 in file S1). The percentage of monocytes with bound platelets was significantly greater in IPF patients following stimulation with 1 µM ADP (29.4%±4.1 (mean ± SEM) versus 16.1%±1.5 in controls; p = <0.01) and 10 µM ADP (44.8%±3.3 versus 32.1%±3.5; p = 0.01). Platelet-monocyte aggregate formation in blood from IPF patients was also significantly increased following stimulation with 5 µM and 10 µM TFLLR (figure 1) when compared to controls. Under basal (unstimulated) conditions, blood from IPF patients exhibited greater platelet-monocyte aggregate formation than controls but this did not reach statistical significance (18.2%±3.9 versus 13.7%±1.4; p = 0.3). The calcium-dependent nature of the interaction between platelets and monocytes was confirmed by addition of EDTA, which resulted in a marked reduction in aggregate formation that did not increase in response to agonist stimulation and did not vary significantly between the groups (figure 2).

Bottom Line: Platelet P-selectin expression was significantly greater in IPF patients than controls following stimulation with 0.1 µM ADP (1.9% positive ±0.5 (mean ± SEM) versus 0.7%±0.1; p = 0.03), 1 µM ADP (9.8%±1.3 versus 3.3%±0.8; p<0.01) and 10 µM ADP (41.3%±4.2 versus 22.5%±2.6; p<0.01).Re-suspension of control platelets in plasma taken from subjects with IPF resulted in increased platelet activation compared to control plasma.IPF patients exhibit increased platelet reactivity compared with controls.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular and Metabolic Research, Hull York Medical School, Cottingham, United Kingdom.

ABSTRACT

Introduction: Idiopathic Pulmonary Fibrosis (IPF) is a progressive, incurable fibrotic interstitial lung disease with a prognosis worse than many cancers. Its pathogenesis is poorly understood. Activated platelets can release pro-fibrotic mediators that have the potential to contribute to lung fibrosis. We determine platelet reactivity in subjects with IPF compared to age-matched controls.

Methods: Whole blood flow cytometry was used to measure platelet-monocyte aggregate formation, platelet P-selectin expression and platelet fibrinogen binding at basal levels and following stimulation with platelet agonists. A plasma swap approach was used to assess the effect of IPF plasma on control platelets.

Results: Subjects with IPF showed greater platelet reactivity than controls. Platelet P-selectin expression was significantly greater in IPF patients than controls following stimulation with 0.1 µM ADP (1.9% positive ±0.5 (mean ± SEM) versus 0.7%±0.1; p = 0.03), 1 µM ADP (9.8%±1.3 versus 3.3%±0.8; p<0.01) and 10 µM ADP (41.3%±4.2 versus 22.5%±2.6; p<0.01). Platelet fibrinogen binding was also increased, and platelet activation resulted in increased platelet-monocyte aggregate formation in IPF patients. Re-suspension of control platelets in plasma taken from subjects with IPF resulted in increased platelet activation compared to control plasma.

Conclusions: IPF patients exhibit increased platelet reactivity compared with controls. This hyperactivity may result from the plasma environment since control platelets exhibit increased activation when exposed to IPF plasma.

Show MeSH
Related in: MedlinePlus