Expression of B-Cell Surface Antigens in Subpopulations of Exosomes Released From B-Cell Lymphoma Cells

Oksvold, Morten P.; Kullmann, A.; Forfang, Lise; Kierulf, Bente; Li, Mu; Brech, Andreas; Vlassov, Alexander V.; Smeland, Erlend B.; Neurauter, Axl; Pedersen, Ketil W.

doi:10.1016/j.clinthera.2014.05.010

Cited by 138 publications

(116 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some groups have recently performed comparative proteomic analyses of medium-sized and small EVs (with or without floatation into a gradient) from platelets (19) or tumor cells (26)(27)(28)(29). Another group has compared the expression of a few surface proteins on B lymphoma-EVs immunoprecipitated with either CD9, CD63, or CD81 (30). Our results are consistent with these other recent works, especially in identifying sEV-enriched proteins (e.g., TSG101 and CD81).…”

Section: Discussionsupporting

confidence: 90%

See 1 more Smart Citation

Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Kowal

Arras

Colombo

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

2,781

188

2,965

View full text Add to dashboard Cite

Extracellular vesicles (EVs) have become the focus of rising interest because of their numerous functions in physiology and pathology. Cells release heterogeneous vesicles of different sizes and intracellular origins, including small EVs formed inside endosomal compartments (i.e., exosomes) and EVs of various sizes budding from the plasma membrane. Specific markers for the analysis and isolation of different EV populations are missing, imposing important limitations to understanding EV functions. Here, EVs from human dendritic cells were first separated by their sedimentation speed, and then either by their behavior upon upward floatation into iodixanol gradients or by immuno-isolation. Extensive quantitative proteomic analysis allowing comparison of the isolated populations showed that several classically used exosome markers, like major histocompatibility complex, flotillin, and heatshock 70-kDa proteins, are similarly present in all EVs. We identified proteins specifically enriched in small EVs, and define a set of five protein categories displaying different relative abundance in distinct EV populations. We demonstrate the presence of exosomal and nonexosomal subpopulations within small EVs, and propose their differential separation by immuno-isolation using either CD63, CD81, or CD9. Our work thus provides guidelines to define subtypes of EVs for future functional studies.exosomes | microvesicles | ectosomes | dendritic cells | extracellular vesicles

show abstract

Section: Discussionsupporting

confidence: 90%

“…Some cell types, however, secrete sEVs devoid of CD63 (30,33,34), and for these it will be necessary to analyze instead either CD81-or CD9-bearing sEVs. In addition, given its presence in large EVs, CD63 cannot be used as specific exosome-isolation tool without a first step of elimination of these large EVs.…”

Section: Discussionmentioning

confidence: 99%

Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Kowal

Arras

Colombo

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

2,781

188

2,965

View full text Add to dashboard Cite

show abstract

“…Size-based separation cannot distinguish among subpopulations of EVs. For example, if the goal is to distinguish the CD81+ subpopulation of EVs from the CD63+ subpopulation, immunoaffinity is recommended [64]. Many studies have discovered EV biogenesis and subpopulations by exploiting the interaction of surface proteins with specific antibodies.…”

Section: Immunoaffinitymentioning

confidence: 99%

Methods to isolate extracellular vesicles for diagnosis

Kang

Kim

Park

2017

Micro and Nano Syst Lett

View full text Add to dashboard Cite

Extracellular vesicles (EVs) are small membrane-bound bodies that are released into extracellular space by diverse cells, and are found in body fluids like blood, urine and saliva. EVs contain RNA, DNA and proteins, which can be biomarkers for diagnosis. EVs can be obtained by minimally-invasive biopsy, so they are useful in disease diagnosis. High yield and purity contribute to precise diagnosis of disease, but damaged EVs and impurities can cause confused results. However, EV isolation methods have different yields and purities. Furthermore, the isolation method that is most suitable to maximize EV recovery efficiency depends on the experimental conditions. This review focuses on merits and demerits of several types of EV isolation methods, and provides examples of how to diagnose disease by exploiting information obtained by analysis of EVs.

show abstract

“…Because bulk assays analyze an ensemble of EV, bulk assays are generally more sensitive and faster than single EV techniques. A typical disadvantage of bulk assays is that information on the EV concentration, biochemical heterogeneity, and polydispersity is lost [1,2]. We will limit the scope of this review by exclusion of proteomics, lipidomics, nucleotide sequencing assays, and polymerase chain reaction [3][4][5][6][7][8], enabling us to focus on bulk immunoassays (BIA).…”

Section: Introductionmentioning

confidence: 99%

Bulk immunoassays for analysis of extracellular vesicles

2017

View full text Add to dashboard Cite

There is increasing clinical interest in extracellular vesicles (EV) for diagnostic and treatment purposes. This review provides an overview of bulk immunoassays to analyse EV. Western blot and enzyme-linked immunosorbent assay are still the two predominant bulk immunoassays. Recently, new assays have become available that can detect exposure to EV concentrations that are up to 10,000-fold lower. This is advantageous for applications that detect rare EV. Other important parameters are the detectable concentration range, the required sample volume, whether simultaneous presence of different antigens on a single EV can be detected, size selectivity of each assay and practical considerations. In this review, we will explain the working principles of the traditional and novel assays together with their performance parameters. The most sensitive assays are micro-nuclear magnetic resonance, surface plasmon resonance, and time-resolved fluorescent immunoassay.

show abstract

Expression of B-Cell Surface Antigens in Subpopulations of Exosomes Released From B-Cell Lymphoma Cells

Cited by 138 publications

References 54 publications

Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Methods to isolate extracellular vesicles for diagnosis

Bulk immunoassays for analysis of extracellular vesicles

Contact Info

Product

Resources

About