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Extensive surface protein profiles of extracellular vesicles from cancer cells may provide diagnostic signatures from blood samples.

Belov L, Matic KJ, Hallal S, Best OG, Mulligan SP, Christopherson RI - J Extracell Vesicles (2016)

Bottom Line: Biotinylated antibodies specific for EpCAM (CD326) and CD19, respectively, were used to detect captured particles by enhanced chemiluminescence.These EV expressed a subset (~40%) of the proteins detected on CLL cells from the same patients: moderate or high levels of CD5, CD19, CD31, CD44, CD55, CD62L, CD82, HLA-A,B,C, HLA-DR; low levels of CD21, CD49c, CD63.None of these proteins was detected on EV from the plasma of age- and gender-matched healthy individuals.

View Article: PubMed Central - PubMed

Affiliation: School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia; larissa.belov@sydney.edu.au.

ABSTRACT
Extracellular vesicles (EV) are membranous particles (30-1,000 nm in diameter) secreted by cells. Important biological functions have been attributed to 2 subsets of EV, the exosomes (bud from endosomal membranes) and the microvesicles (MV; bud from plasma membranes). Since both types of particles contain surface proteins derived from their cell of origin, their detection in blood may enable diagnosis and prognosis of disease. We have used an antibody microarray (DotScan) to compare the surface protein profiles of live cancer cells with those of their EV, based on their binding patterns to immobilized antibodies. Initially, EV derived from the cancer cell lines, LIM1215 (colorectal cancer) and MEC1 (B-cell chronic lymphocytic leukaemia; CLL), were used for assay optimization. Biotinylated antibodies specific for EpCAM (CD326) and CD19, respectively, were used to detect captured particles by enhanced chemiluminescence. Subsequently, this approach was used to profile CD19(+) EV from the plasma of CLL patients. These EV expressed a subset (~40%) of the proteins detected on CLL cells from the same patients: moderate or high levels of CD5, CD19, CD31, CD44, CD55, CD62L, CD82, HLA-A,B,C, HLA-DR; low levels of CD21, CD49c, CD63. None of these proteins was detected on EV from the plasma of age- and gender-matched healthy individuals.

No MeSH data available.


Related in: MedlinePlus

DotScan profiling of MEC1 cells (b) and their EV (c). The key (a) shows locations of antibodies (as for Fig. 1), with shaded antibodies indicating cell capture. Detection of captured cells was by optical scanning (b). EV (7.35×1010) were detected by ECL using biotinylated CD19 antibody, with a 5 min exposure on ECL film (c). NanoSight analysis shows the size distribution of MEC1 EV (d). A Venn diagram (e) compares surface profiles of MEC1 cells with their EV. The number above the peak represents mode size in nm.
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Figure 0002: DotScan profiling of MEC1 cells (b) and their EV (c). The key (a) shows locations of antibodies (as for Fig. 1), with shaded antibodies indicating cell capture. Detection of captured cells was by optical scanning (b). EV (7.35×1010) were detected by ECL using biotinylated CD19 antibody, with a 5 min exposure on ECL film (c). NanoSight analysis shows the size distribution of MEC1 EV (d). A Venn diagram (e) compares surface profiles of MEC1 cells with their EV. The number above the peak represents mode size in nm.

Mentions: Figure 2 shows a comparison of the surface profiles of MEC1 cells (Fig. 2b) and their EV (Fig. 2c). NanoSight analysis of MEC1 EV (Fig. 2d) showed particles with a mode size of 114 nm. Of the 36 proteins detected on the cells, 25 (69.4%) were also detected on their EV, as summarized in the Venn diagram (Fig. 2e). CD15 and CD31 were detected on the MEC1 EV, but not the cells.


Extensive surface protein profiles of extracellular vesicles from cancer cells may provide diagnostic signatures from blood samples.

Belov L, Matic KJ, Hallal S, Best OG, Mulligan SP, Christopherson RI - J Extracell Vesicles (2016)

DotScan profiling of MEC1 cells (b) and their EV (c). The key (a) shows locations of antibodies (as for Fig. 1), with shaded antibodies indicating cell capture. Detection of captured cells was by optical scanning (b). EV (7.35×1010) were detected by ECL using biotinylated CD19 antibody, with a 5 min exposure on ECL film (c). NanoSight analysis shows the size distribution of MEC1 EV (d). A Venn diagram (e) compares surface profiles of MEC1 cells with their EV. The number above the peak represents mode size in nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834364&req=5

Figure 0002: DotScan profiling of MEC1 cells (b) and their EV (c). The key (a) shows locations of antibodies (as for Fig. 1), with shaded antibodies indicating cell capture. Detection of captured cells was by optical scanning (b). EV (7.35×1010) were detected by ECL using biotinylated CD19 antibody, with a 5 min exposure on ECL film (c). NanoSight analysis shows the size distribution of MEC1 EV (d). A Venn diagram (e) compares surface profiles of MEC1 cells with their EV. The number above the peak represents mode size in nm.
Mentions: Figure 2 shows a comparison of the surface profiles of MEC1 cells (Fig. 2b) and their EV (Fig. 2c). NanoSight analysis of MEC1 EV (Fig. 2d) showed particles with a mode size of 114 nm. Of the 36 proteins detected on the cells, 25 (69.4%) were also detected on their EV, as summarized in the Venn diagram (Fig. 2e). CD15 and CD31 were detected on the MEC1 EV, but not the cells.

Bottom Line: Biotinylated antibodies specific for EpCAM (CD326) and CD19, respectively, were used to detect captured particles by enhanced chemiluminescence.These EV expressed a subset (~40%) of the proteins detected on CLL cells from the same patients: moderate or high levels of CD5, CD19, CD31, CD44, CD55, CD62L, CD82, HLA-A,B,C, HLA-DR; low levels of CD21, CD49c, CD63.None of these proteins was detected on EV from the plasma of age- and gender-matched healthy individuals.

View Article: PubMed Central - PubMed

Affiliation: School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia; larissa.belov@sydney.edu.au.

ABSTRACT
Extracellular vesicles (EV) are membranous particles (30-1,000 nm in diameter) secreted by cells. Important biological functions have been attributed to 2 subsets of EV, the exosomes (bud from endosomal membranes) and the microvesicles (MV; bud from plasma membranes). Since both types of particles contain surface proteins derived from their cell of origin, their detection in blood may enable diagnosis and prognosis of disease. We have used an antibody microarray (DotScan) to compare the surface protein profiles of live cancer cells with those of their EV, based on their binding patterns to immobilized antibodies. Initially, EV derived from the cancer cell lines, LIM1215 (colorectal cancer) and MEC1 (B-cell chronic lymphocytic leukaemia; CLL), were used for assay optimization. Biotinylated antibodies specific for EpCAM (CD326) and CD19, respectively, were used to detect captured particles by enhanced chemiluminescence. Subsequently, this approach was used to profile CD19(+) EV from the plasma of CLL patients. These EV expressed a subset (~40%) of the proteins detected on CLL cells from the same patients: moderate or high levels of CD5, CD19, CD31, CD44, CD55, CD62L, CD82, HLA-A,B,C, HLA-DR; low levels of CD21, CD49c, CD63. None of these proteins was detected on EV from the plasma of age- and gender-matched healthy individuals.

No MeSH data available.


Related in: MedlinePlus