Abstract 20: Resolution Phenotype of Monocytes and Macrophages is Altered in Peripheral Arterial Disease

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Melinda S Schaller ◽  
Laura Menke ◽  
Mian Chen ◽  
Warren J Gasper ◽  
S. Marlene Grenon ◽  
...  
Keyword(s):  
Peripheral Arterial Disease ◽  
Cell Surface ◽  
Phagocytic Activity ◽  
Mononuclear Cells ◽  
Gradient Centrifugation ◽  
Arterial Disease ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Peripheral Blood Mononuclear ◽  
Peripheral Arterial

Introduction: Peripheral arterial disease (PAD) is a chronic disease characterized by systemic inflammation. Monocytes (Mo) and macrophages play a central role in vascular inflammation and its resolution. We hypothesize that impaired resolution in PAD results in poor clinical outcomes. Methods: Resolution phenotype was assessed by phagocytic activity of leukocytes, Mo cell surface markers, and cytokine profiling of Mo-derived macrophages (MDM). Phagocytosis and cell-surface markers were determined by flow cytometry. MDMs were generated from peripheral blood mononuclear cells via density gradient centrifugation. Cytokines were measured by ELISA following MDM differentiation and subsequent stimulation with LPS. Results: Circulating Mo and neutrophils (PMN) isolated from PAD patients (n=9) demonstrated significantly lower phagocytic activity (Mo: >30%, p<.001; PMN: >25%, p<.01, Fig. 1) as compared to healthy subjects (HS) (n=14). Cell-surface marker analysis demonstrated a higher proportion of the pro-inflammatory intermediate Mo subset (CD14 ++ 16 + , 1.8-fold, p=.04) in PAD compared to HS. MDM from PAD subjects retain their intrinsic inflammatory program by producing more IL-6 (PAD 3138±2676 ng/mL, HS 731±854 ng/mL p=.03) and IL-1β (PAD 244±236 ng/mL, HS 24.1±23.8 ng/mL p=.04) than those from HS. Upon stimulation with LPS, MDM from PAD subjects secrete more IL-6 (PAD 23353±22483 ng/mL, HS 5097±5836 ng/mL p=.05) than those from HS. Conclusions: Circulating Mo and PMN in patients with PAD have substantially lower phagocytic activity as well as a greater proportion of the pro-inflammatory intermediate Mo subset compared to HS. MDM preserve their elevated inflammatory state throughout culture and retain a heightened response upon latter stimulatory cues. Collectively these data demonstrate a heightened inflammatory and impaired resolution phenotype in PAD that has potential implications for disease progression and response to interventions.

Phenotypic characterisation of pro-inflammatory monocytes and dendritic cells in peripheral arterial disease

2012 ◽  
Vol 108 (12) ◽  
pp. 1198-1207 ◽  
Author(s):  
Viviane Obst ◽  
Christopher Doppler ◽  
Marie-Christine Radmacher ◽  
Martin Scheer ◽  
Markus Radsak ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Peripheral Arterial Disease ◽  
Intermittent Claudication ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Arterial Disease ◽  
Regulatory Function ◽  
Inflammatory Monocytes ◽  
Phenotypic Characterisation ◽  
Peripheral Arterial

SummaryAtherosclerosis is a chronic inflammatory process involving antigen-presenting cells like monocytes and dendritic cells (DC). The aim of this study was to perform a phenotypic characterisation of these cell types in patients with different degrees of peripheral arterial disease (PAD). Sixty patients with PAD [N= 30 intermittent claudication (IC), N= 30 critical limb ischemia (CLI)] and 30 controls were included. Peripheral blood leucocytes were analysed from peripheral blood by flow cytometry using different gating strategies to directly identify and analyse monocytes, myeloid DC, (mDC) and plasmacytoid DC (pDC). PAD patients showed a significantly higher proportion of proinflammatory CD14++CD16+ monocytes (p<0.0001) compared with healthy individuals. We found an increased number of mDC/ml and a reduced number of pDC/ml (both p<0.01) in PAD patients, leading to a shift in the mDC/pDC ratio (p<0.01). As compared to patients with intermittent claudication, CLI patients presented a reduced expression of HLA-DR (p<0.01), CD86 and CD40 on both mDCs and pDCs (p<0.01). Peripheral blood monocytes show a proinflammatory phenotype in PAD patients compared to controls. In contrast, CLI patients show a reduced expression of proinflammatory markers. We hypothesise that severe ischaemia and/or prolonged inflammation in CLI might lead to a paradoxical attenuation in the proinflammatory membrane pattern of circulating mononuclear cells, possibly hindering an adequate regulatory function of mDCs and pDCs and favouring the progression of disease.

No Clinical Benefit of Intramuscular Delivery of Bone Marrow-derived Mononuclear Cells in Nonreconstructable Peripheral Arterial Disease

2018 ◽  
Vol 268 (5) ◽  
pp. 756-761
Author(s):  
Jan H. N. Lindeman ◽  
Jaap Jan Zwaginga ◽  
Graziella Kallenberg-Lantrua ◽  
Rob C. van Wissen ◽  
Abbey Schepers ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Arterial Disease ◽  
Clinical Benefit ◽  
Mononuclear Cells ◽  
Arterial Disease ◽  
Peripheral Arterial ◽  
Intramuscular Delivery

scholarly journals Percutaneous Transluminal Angioplasty in Patients with Peripheral Arterial Disease Does Not Affect Circulating Monocyte Subpopulations

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Pawel Maga ◽  
Tomasz P. Mikolajczyk ◽  
Lukasz Partyka ◽  
Marek Krzanowski ◽  
Krzysztof P. Malinowski ◽  
...  
Keyword(s):  
Peripheral Arterial Disease ◽  
Percutaneous Transluminal Angioplasty ◽  
Mononuclear Cells ◽  
Arterial Disease ◽  
Transluminal Angioplasty ◽  
Observation Study ◽  
Surface Receptors ◽  
Monocyte Subpopulations ◽  
Peripheral Arterial ◽  
Percutaneous Transluminal

Monocytes are mononuclear cells characterized by distinct morphology and expression of CD14 and CD16 surface receptors. Classical, quiescent monocytes are positive for CD14 (lipopolysaccharide receptor) but do not express Fc gamma receptor III (CD16). Intermediate monocytes coexpress CD16 and CD14. Nonclassical monocytes with low expression of CD14 represent mature macrophage-like monocytes. Monocyte behavior in peripheral arterial disease (PAD) and during vessel wall directed treatment is not well defined. This observation study aimed at monitoring of acute changes in monocyte subpopulations during percutaneous transluminal angioplasty (PTA) in PAD patients. Patients with Rutherford 3 and 4 PAD with no signs of inflammatory process underwent PTA of iliac, femoral, or popliteal segments. Flow cytometry for CD14, CD16, HLA-DR, CD11b, CD11c, and CD45RA antigens allowed characterization of monocyte subpopulations in blood sampled before and after PTA (direct angioplasty catheter sampling). Patients were clinically followed up for 12 months. All 61 enrolled patients completed 12-month follow-up. Target vessel failure occurred in 12 patients. While absolute counts of monocyte were significantly lower after PTA, only subtle monocyte activation after PTA (CD45RA andβ-integrins) occurred. None of the monocyte parameters correlated with long-term adverse clinical outcome. Changes in absolute monocyte counts and subtle changes towards an activation phenotype after PTA may reflect local cell adhesion phenomenon in patients with Rutherford 3 or 4 peripheral arterial disease.

Abstract 17327: Ischemic Skeletal Muscle From Patients With Peripheral Arterial Disease Displays Marked Desialylation of Cell Surface Glycans: Implications for Ischemia-Targeted Gene Therapy

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Arabindra B Katwal ◽  
Eric T Mulkey ◽  
R John Lye ◽  
Brent A French ◽  
Brian H Annex
Keyword(s):  
Skeletal Muscle ◽  
Gene Therapy ◽  
Peripheral Arterial Disease ◽  
Cell Surface ◽  
Limb Ischemia ◽  
Arterial Disease ◽  
Fluorescence Signal ◽  
Differential Distribution ◽  
Peripheral Arterial ◽  
Cell Surface Glycans

Introduction: Ischemic skeletal muscle is preferentially targeted after IV injection of AAV9 in mouse models of hind-limb ischemia (HLI). Ischemia is known to induce desialylation of cell surface glycans, unmasking the cellular receptor for AAV9 and resulting in significantly higher levels of selective gene expression in ischemic vs non-ischemic muscle. To assess clinical relevance, we compared the differential distribution of sialylated and desialylated cell surface glycans in skeletal muscle biopsies from peripheral arterial disease (PAD) patients and normal controls. Methods: Gastrocnemius (GA) muscle samples from PAD patients and normals (n=29 each) were biopsied under IRB-approved protocols. Muscle sections were stained using the lectins MAL-I and ECL. MAL-I binds to α2,3-sialylated glycans whereas ECL binds to desialylated galactose residues on cell surface glycans. Lectins were labeled with Streptavidin-Alexa-488 (red) for MAL-I and SA-555 (green) for ECL, and visualized using fluorescence microscopy. Quantification of mean fluorescence signal intensity of lectin staining was performed on each section. Results: Visually, MAL-I staining was markedly higher and ECL staining lower in non-ischemic GA from normals when compared to ischemic GA from PAD patients (Figure). The ratio of MAL-I to ECL integrated fluorescence intensity in the ischemic muscles from PAD patients (1.13 ± 0.13) was significantly lower than the same ratio in non-ischemic muscles from normals (1.58 ± 0.14, p<0.02). Conclusions: Desialylation of cell surface glycans is significantly higher in ischemic skeletal muscle from PAD patients, consistent with previous work in a mouse model of HLI. This desialylation can potentially be exploited to treat human PAD with gene therapy, since these desialylated cell surface glycans serve as cell surface receptors for AAV9 and provide for efficient and selective gene transfer to ischemic skeletal muscle following systemic or local delivery.

Management of Peripheral Arterial Disease

2006 ◽  
Vol 39 (3) ◽  
pp. 44
Author(s):  
WILLIAM E. GOLDEN ◽  
ROBERT H. HOPKINS
Keyword(s):  
Peripheral Arterial Disease ◽  
Arterial Disease ◽  
Peripheral Arterial
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