Role of Proinflammatory CD68
            <sup>+</sup>
            Mannose Receptor
            <sup>−</sup>
            Macrophages in Peroxiredoxin-1 Expression and in Abdominal Aortic Aneurysms in Humans

Boytard, Ludovic; Spear, Rafaëlle; Chinetti-Gbaguidi, Giulia; Acosta‐Martin, Adelina E.; Vanhoutte, Jonathan; Lamblin, Nicolas; Staels, Bart; Amouyel, Philippe; Haulon, Stéphan; Pinet, Florence

doi:10.1161/atvbaha.112.300663

Cited by 70 publications

(72 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Clearly, we observed a distinct morphology of macrophages M1 and M2 after 12 days of primary culture. As previously published, we verified by RT-PCR the presence of mRNA coding for TNF and IL1B in M1 macrophages and for MRC1 and CCL18 in M2 macrophages 14 (not shown).…”

Section: Representative Resultssupporting

confidence: 81%

Proteomic Profiling of Macrophages by 2D Electrophoresis

Bouvet¹,

Turkieh²,

Acosta‐Martin³

et al. 2014

JoVE

Self Cite

View full text Add to dashboard Cite

The goal of the two-dimensional (2D) electrophoresis protocol described here is to show how to analyse the phenotype of human cultured macrophages. The key role of macrophages has been shown in various pathological disorders such as inflammatory, immunological, and infectious diseases. In this protocol, we use primary cultures of human monocyte-derived macrophages that can be differentiated into the M1 (pro-inflammatory) or the M2 (anti-inflammatory) phenotype. This in vitro model is reliable for studying the biological activities of M1 and M2 macrophages and also for a proteomic approach. Proteomic techniques are useful for comparing the phenotype and behaviour of M1 and M2 macrophages during host pathogenicity. 2D gel electrophoresis is a powerful proteomic technique for mapping large numbers of proteins or polypeptides simultaneously. We describe the protocol of 2D electrophoresis using fluorescent dyes, named 2D Differential Gel Electrophoresis (DIGE). The M1 and M2 macrophages proteins are labelled with cyanine dyes before separation by isoelectric focusing, according to their isoelectric point in the first dimension, and their molecular mass, in the second dimension. Separated protein or polypeptidic spots are then used to detect differences in protein or polypeptide expression levels. The proteomic approaches described here allows the investigation of the macrophage protein changes associated with various disorders like host pathogenicity or microbial toxins. Video LinkThe video component of this article can be found at

show abstract

Section: Representative Resultssupporting

confidence: 81%

Proteomic Profiling of Macrophages by 2D Electrophoresis

Bouvet¹,

Turkieh²,

Acosta‐Martin³

et al. 2014

JoVE

Self Cite

View full text Add to dashboard Cite

show abstract

“…[24]. In the adventitia, B lymphocytes and mast cells predominated, together with the proinflammatory CD68 + MR − (Mannose Receptor) macrophage subtype as previously shown [10] and neutrophils and T lymphocytes predominated, together with the anti-inflammatory subtype CD68 + MR + macrophages as previously shown in the intraluminal thrombus [10]. …”

Section: Resultsmentioning

confidence: 59%

“…Numerous inflammatory cells are involved in AAA formation and growth [7,9]. Recently, we showed that the two subtypes of macrophages, M1 with proinflammatory properties and M2, with anti-inflammatory properties are distributed differently in the aneurysmal wall [10]. Neutrophils are known to be involved in AAA [11] by recruiting other inflammatory cells [9] or producing MMPs such as MMP9 [12].…”

Section: Introductionmentioning

confidence: 99%

“…We therefore first analyzed the specific distribution of inflammatory cells (neutrophils, B and T lymphocytes and mast cells) as we did previously for the two subtypes of macrophages [10] in the different layers of AAA. The expression of the selected miRNAs detected in ATLOs was then evaluated by quantitative RT-PCR (qRT-PCR) in LCM-isolated ATLOs, M1 and M2 macrophages as well as in entire aneurysmal and control aorta samples.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Adventitial Tertiary Lymphoid Organs as Potential Source of MicroRNA Biomarkers for Abdominal Aortic Aneurysm

Spear

Boytard

Blervaque

et al. 2015

IJMS

Self Cite

View full text Add to dashboard Cite

Abdominal aortic aneurysm (AAA) is an inflammatory disease associated with marked changes in the cellular composition of the aortic wall. This study aims to identify microRNA (miRNA) expression in aneurysmal inflammatory cells isolated by laser microdissection from human tissue samples. The distribution of inflammatory cells (neutrophils, B and T lymphocytes, mast cells) was evaluated in human AAA biopsies. We observed in half of the samples that adventitial tertiary lymphoid organs (ATLOs) with a thickness from 0.5 to 2 mm were located exclusively in the adventitia. Out of the 850 miRNA that were screened by microarray in isolated ATLOs (n = 2), 164 miRNAs were detected in ATLOs. The three miRNAs (miR-15a-3p, miR-30a-5p and miR-489-3p) with the highest expression levels were chosen and their expression quantified by RT-PCR in isolated ATLOs (n = 4), M1 (n = 2) and M2 macrophages (n = 2) and entire aneurysmal biopsies (n = 3). Except for the miR-30a-5p, a similar modulation was found in ATLOs and the two subtypes of macrophages. The modulated miRNAs were then evaluated in the plasma of AAA patients for their potential as AAA biomarkers. Our data emphasize the potential of miR-15a-3p and miR-30a-5p as biomarkers of AAA but also as triggers of ATLO evolution. Further investigations will be required to evaluate their targets in order to better understand AAA pathophysiology.

show abstract

“…Many clinical applications have been investigated by this method, including biomarker identification of breast cancer (Thakur et al, 2011;Zanni and Chan, 2011;Zhang et al, 2005), pancreatic ductal carcinoma (Shekouh et al, 2003), nasopharyngeal carcinoma (Cheng et al, 2008), hepatocellular carcinoma (Wong and Luk, 2012), and abdominal aortic aneurysms (Boytard et al, 2013). In this last study, Boytard and colleagues highlighted that CD68( + )MR(-) macrophages could contribute to aneurysmal pathology, on the basis of the analyses of 2DE separated proteins originated from different types of macrophages.…”

Section: Laser Capture Microdissection (Lcm)mentioning

confidence: 99%

Tissue Proteomics for the Next Decade? Towards a Molecular Dimension in Histology

Longuespée

Fléron

Pottier

et al. 2014

OMICS: A Journal of Integrative Biology

View full text Add to dashboard Cite

show abstract

Role of Proinflammatory CD68 ⁺ Mannose Receptor ⁻ Macrophages in Peroxiredoxin-1 Expression and in Abdominal Aortic Aneurysms in Humans

Cited by 70 publications

References 47 publications

Proteomic Profiling of Macrophages by 2D Electrophoresis

Proteomic Profiling of Macrophages by 2D Electrophoresis

Adventitial Tertiary Lymphoid Organs as Potential Source of MicroRNA Biomarkers for Abdominal Aortic Aneurysm

Tissue Proteomics for the Next Decade? Towards a Molecular Dimension in Histology

Contact Info

Product

Resources

About

Role of Proinflammatory CD68 + Mannose Receptor − Macrophages in Peroxiredoxin-1 Expression and in Abdominal Aortic Aneurysms in Humans

Cited by 70 publications

References 47 publications

Proteomic Profiling of Macrophages by 2D Electrophoresis

Proteomic Profiling of Macrophages by 2D Electrophoresis

Adventitial Tertiary Lymphoid Organs as Potential Source of MicroRNA Biomarkers for Abdominal Aortic Aneurysm

Tissue Proteomics for the Next Decade? Towards a Molecular Dimension in Histology

Contact Info

Product

Resources

About

Role of Proinflammatory CD68 ⁺ Mannose Receptor ⁻ Macrophages in Peroxiredoxin-1 Expression and in Abdominal Aortic Aneurysms in Humans