Proteomic characterization of thymocyte-derived microvesicles and apoptotic bodies in BALB/c mice

Turiák, Lilla; Misják, Petra; Szabó, Tamás; Aradi, Borbála; Pálóczi, Krisztina; Ozohanics, Olivér; Drahos, László; Kittel, Ágnes; Falus, András; Buzás, Edit I.; Vékey, Károly

doi:10.1016/j.jprot.2011.05.023

Cited by 130 publications

(107 citation statements)

References 35 publications

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“…*P , 0.05 compared with control (24 h). ORIGINAL RESEARCH structure and the size heterogeneity of the MPs generated from lung ECs are similar to MPs reported for human umbilical vein ECs (33), Jurkat cells (34), and thymocytes (35). Despite the significant association of MPs with inflammatory diseases, few studies have addressed MP participation in the pathogenesis of ALI/VILI or as markers of lung injury.…”

Section: Anti-cd31supporting

confidence: 63%

Pathologic Mechanical Stress and Endotoxin Exposure Increases Lung Endothelial Microparticle Shedding

Letsiou¹,

Sammani

Zhang

et al. 2015

Am J Respir Cell Mol Biol

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Acute lung injury (ALI) results from infectious challenges and from pathologic lung distention produced by excessive tidal volume delivered during mechanical ventilation (ventilator-induced lung injury [VILI]) and is characterized by extensive alveolar and vascular dysfunction. Identification of novel ALI therapies is hampered by the lack of effective ALI/VILI biomarkers. We explored endothelial cell (EC)-derived microparticles (EMPs) (0.1-1 mm) as potentially important markers and potential mediators of lung vascular injury in preclinical models of ALI and VILI. We characterized EMPs (annexin V and CD31 immunoreactivity) produced from human lung ECs exposed to physiologic or pathologic mechanical stress (5 or 18% cyclic stretch [CS]) or to endotoxin (LPS). EC exposure to 18% CS or to LPS resulted in increased EMP shedding compared with static cells (z 4-fold and z 2.5-fold increases, respectively). Proteomic analysis revealed unique 18% CS-derived (n = 10) and LPS-derived EMP proteins (n = 43). VILI-challenged mice (40 ml/kg, 4 h) exhibited increased plasma and bronchoalveolar lavage CD62E (E-selectin)-positive MPs compared with control mice. Finally, mice receiving intratracheal instillation of 18% CS-derived EMPs displayed significant lung inflammation and injury. These findings indicate that ALI/VILI-producing stimuli induce significant shedding of distinct EMP populations that may serve as potential ALI biomarkers and contribute to the severity of lung injury.Keywords: microparticles; ventilator-induced lung injury; cyclic stretch; LPS; proteomics Clinical RelevanceAcute lung injury (ALI) results from infectious challenges and pathologic lung distention produced by excessive tidal volume delivered during mechanical ventilation (ventilator-induced lung injury [VILI]) and is characterized by extensive alveolar and vascular dysfunction. Identification of novel ALI therapies is hampered by the lack of effective ALI/VILI biomarkers. We explored endothelial cell (EC)-derived microparticles (EMPs) (0.1-1 mm) as potentially important markers and potential mediators of lung vascular injury in preclinical models of ALI and VILI. Our findings indicate that ALI/VILIproducing stimuli induce significant shedding of distinct EMP populations that may serve as potential ALI biomarkers and contribute to the severity of lung injury.

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Section: Anti-cd31supporting

confidence: 63%

Pathologic Mechanical Stress and Endotoxin Exposure Increases Lung Endothelial Microparticle Shedding

Letsiou¹,

Sammani

Zhang

et al. 2015

Am J Respir Cell Mol Biol

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“…The major difference between apoptotic vesicles and other cell-derived vesicles is their size. In all studies so far, the diameter of apoptotic vesicles is reported to range between 1 and 5 m (Kerr et al, 1972;Hristov et al, 2004;Turiá k et al, 2011). This is in the precise size range of platelets in the human blood.…”

Section: Apoptotic Vesiclesmentioning

confidence: 88%

Classification, Functions, and Clinical Relevance of Extracellular Vesicles

et al. 2012

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“…Cell viability was >90-95% as confirmed by using annexin V FITC and propidium iodide (both from BD Biosciences) staining as described before. 3,12 EV-containing conditioned serum-free media were subjected to centrifugation at 300g to deplete cells, and stored at −80°C prior to analysis by TRPS. For transmission electron microscopy and flow cytometry three different EV subpopulations were isolated including APOs, MVs, and EXOs by size filtration and differential centrifugation.…”

Section: Ev Isolation From Cell Culture Supernatantsmentioning

confidence: 99%

Differential detergent sensitivity of extracellular vesicle subpopulations

et al. 2015

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a Extracellular vesicles (including exosomes, microvesicles and apoptotic bodies) are currently attracting rapidly increasing attention from various fields of biology due to their ability to carry complex information and act as autocrine, paracrine and even endocrine intercellular messengers. In the present study we investigated the sensitivity of size-based subpopulations of extracellular vesicles to different concentrations of detergents including sodium dodecyl sulphate, Triton X-100, Tween 20 and deoxycholate. We determined the required detergent concentration that lysed each of the vesicle subpopulations secreted by Jurkat, THP-1, MiaPaCa and U937 human cell lines. We characterized the vesicles by tunable resistive pulse sensing, flow cytometry and transmission electron microscopy. Microvesicles and apoptotic bodies were found to be more sensitive to detergent lysis than exosomes. Furthermore, we found evidence that sodium dodecyl sulphate and Triton X-100 were more effective in vesicle lysis at low concentrations than deoxycholate or Tween 20. Taken together, our data suggest that a combination of differential detergent lysis with tunable resistive pulse sensing or flow cytometry may prove useful for simple and fast differentiation between exosomes and other extracellular vesicle subpopulations as well as between vesicular and non-vesicular structures.

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Proteomic characterization of thymocyte-derived microvesicles and apoptotic bodies in BALB/c mice

Cited by 130 publications

References 35 publications

Pathologic Mechanical Stress and Endotoxin Exposure Increases Lung Endothelial Microparticle Shedding

Pathologic Mechanical Stress and Endotoxin Exposure Increases Lung Endothelial Microparticle Shedding

Classification, Functions, and Clinical Relevance of Extracellular Vesicles

Differential detergent sensitivity of extracellular vesicle subpopulations

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