Physical and functional characterisation of apoptotic cell-derived extracellular vesicles

Lois Hawkins, Khaled Alghareeb, Vinod Nadella, Parbata Chauhan, Charlotte E. Bland, Andrew Devitt

Research output: Contribution to conferencePoster

Abstract

Damaged, aged or unwanted cells are removed from the body by an active process known as apoptosis. This highly orchestrated programme results in the exposure of 'flags' at the dying cell surface and the release of attractive signals to recruit phagocytes. Together these changes ensure efficient phagocytic removal of dying cells and prevention of inflammatory and autoimmune disorders.
Extracellular vesicles (EV) are released from a variety of cells (both viable and apoptotic) and they serve as a novel means of intercellular communication. They range in size: 70-100nm ('exosomes') through 100-1000nm ('microparticles') to large vesicles released from dying cells ('apoptotic bodies'). Release of apoptotic cell-derived extracellular vesicles (acdEV) of less than 1000nm is an important mechanism by which phagocytes are attracted to sites of cell death. Using a variety of approaches we characterize the release, physical characteristics and function of acdEV. Using fluorescence microscopy we demonstrate release of ICAM-3 on acdEV from dying leukocytes and, through the use of resistive pulse technology (qNano, IZON Science), we accurately size and quantitate acdEV release. The function of acdEV is revealed through the use of both horizontal chemotaxis assays (Dunn chambers) and vertical transwell migration assays (Cell-IQ, CM Technologies). These assays reveal potent chemoattractive capacity of acdEV and associated ICAM-3. Additionally we demonstrate an additional novel function of acdEV as anti-inflammatory immune-modulators. These data support an integrated approach to the physical and functional analyses of EV.
LanguageEnglish
Publication statusPublished - 2014
EventBritish Society for Immunology (BSI) annual congress - UK, Brighton, United Kingdom
Duration: 1 Dec 20144 Dec 2014

Congress

CongressBritish Society for Immunology (BSI) annual congress
Abbreviated titleBSI Congress
CountryUnited Kingdom
CityBrighton
Period1/12/144/12/14

Fingerprint

Phagocytes
Cell Migration Assays
Extracellular Vesicles
Exosomes
Technology
Chemotaxis
Fluorescence Microscopy
Leukocytes
Cell Death
Anti-Inflammatory Agents
Apoptosis
Cell Body

Cite this

Hawkins, L., Alghareeb, K., Nadella, V., Chauhan, P., Bland, C. E., & Devitt, A. (2014). Physical and functional characterisation of apoptotic cell-derived extracellular vesicles. Poster session presented at British Society for Immunology (BSI) annual congress, Brighton, United Kingdom.
Hawkins, Lois ; Alghareeb, Khaled ; Nadella, Vinod ; Chauhan, Parbata ; Bland, Charlotte E. ; Devitt, Andrew. / Physical and functional characterisation of apoptotic cell-derived extracellular vesicles. Poster session presented at British Society for Immunology (BSI) annual congress, Brighton, United Kingdom.
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Hawkins, L, Alghareeb, K, Nadella, V, Chauhan, P, Bland, CE & Devitt, A 2014, 'Physical and functional characterisation of apoptotic cell-derived extracellular vesicles' British Society for Immunology (BSI) annual congress, Brighton, United Kingdom, 1/12/14 - 4/12/14, .

Physical and functional characterisation of apoptotic cell-derived extracellular vesicles. / Hawkins, Lois; Alghareeb, Khaled; Nadella, Vinod; Chauhan, Parbata; Bland, Charlotte E.; Devitt, Andrew.

2014. Poster session presented at British Society for Immunology (BSI) annual congress, Brighton, United Kingdom.

Research output: Contribution to conferencePoster

TY - CONF

T1 - Physical and functional characterisation of apoptotic cell-derived extracellular vesicles

AU - Hawkins, Lois

AU - Alghareeb, Khaled

AU - Nadella, Vinod

AU - Chauhan, Parbata

AU - Bland, Charlotte E.

AU - Devitt, Andrew

PY - 2014

Y1 - 2014

N2 - Damaged, aged or unwanted cells are removed from the body by an active process known as apoptosis. This highly orchestrated programme results in the exposure of 'flags' at the dying cell surface and the release of attractive signals to recruit phagocytes. Together these changes ensure efficient phagocytic removal of dying cells and prevention of inflammatory and autoimmune disorders.Extracellular vesicles (EV) are released from a variety of cells (both viable and apoptotic) and they serve as a novel means of intercellular communication. They range in size: 70-100nm ('exosomes') through 100-1000nm ('microparticles') to large vesicles released from dying cells ('apoptotic bodies'). Release of apoptotic cell-derived extracellular vesicles (acdEV) of less than 1000nm is an important mechanism by which phagocytes are attracted to sites of cell death. Using a variety of approaches we characterize the release, physical characteristics and function of acdEV. Using fluorescence microscopy we demonstrate release of ICAM-3 on acdEV from dying leukocytes and, through the use of resistive pulse technology (qNano, IZON Science), we accurately size and quantitate acdEV release. The function of acdEV is revealed through the use of both horizontal chemotaxis assays (Dunn chambers) and vertical transwell migration assays (Cell-IQ, CM Technologies). These assays reveal potent chemoattractive capacity of acdEV and associated ICAM-3. Additionally we demonstrate an additional novel function of acdEV as anti-inflammatory immune-modulators. These data support an integrated approach to the physical and functional analyses of EV.

AB - Damaged, aged or unwanted cells are removed from the body by an active process known as apoptosis. This highly orchestrated programme results in the exposure of 'flags' at the dying cell surface and the release of attractive signals to recruit phagocytes. Together these changes ensure efficient phagocytic removal of dying cells and prevention of inflammatory and autoimmune disorders.Extracellular vesicles (EV) are released from a variety of cells (both viable and apoptotic) and they serve as a novel means of intercellular communication. They range in size: 70-100nm ('exosomes') through 100-1000nm ('microparticles') to large vesicles released from dying cells ('apoptotic bodies'). Release of apoptotic cell-derived extracellular vesicles (acdEV) of less than 1000nm is an important mechanism by which phagocytes are attracted to sites of cell death. Using a variety of approaches we characterize the release, physical characteristics and function of acdEV. Using fluorescence microscopy we demonstrate release of ICAM-3 on acdEV from dying leukocytes and, through the use of resistive pulse technology (qNano, IZON Science), we accurately size and quantitate acdEV release. The function of acdEV is revealed through the use of both horizontal chemotaxis assays (Dunn chambers) and vertical transwell migration assays (Cell-IQ, CM Technologies). These assays reveal potent chemoattractive capacity of acdEV and associated ICAM-3. Additionally we demonstrate an additional novel function of acdEV as anti-inflammatory immune-modulators. These data support an integrated approach to the physical and functional analyses of EV.

M3 - Poster

ER -

Hawkins L, Alghareeb K, Nadella V, Chauhan P, Bland CE, Devitt A. Physical and functional characterisation of apoptotic cell-derived extracellular vesicles. 2014. Poster session presented at British Society for Immunology (BSI) annual congress, Brighton, United Kingdom.