TY - JOUR
T1 - Stimulated release of intraluminal vesicles from Weibel-Palade bodies
AU - Streetley, James
AU - Fonseca, Ana Violeta
AU - Turner, Jack
AU - Kiskin, Nikolai I.
AU - Knipe, Laura
AU - Rosenthal, Peter B.
AU - Carter, Tom
N1 - Funding Information:
The anti-CD63 monoclonal antibody H5C6, developed by J. T. August and J. E. K. Hildreth, was obtained from the Developmental Studies Hybridoma Bank, created by the Eunice Kennedy Shriver National Institute of Child Health and Human Development of the National Institutes of Health and maintained at the Department of Biology at the University of Iowa. T.C. was funded by the UK Medical Research Council under program grant MC_PC_13053. P.B.R. is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001143), the UK Medical Research Council (FC001143), and the Wellcome Trust (FC001143).
Funding Information:
T.C. was funded by the UK Medical Research Council under program grant MC_PC_13053. P.B.R. is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001143), the UK Medical Research Council (FC001143), and the Wellcome Trust (FC001143).
Publisher Copyright:
© 2019 by The American Society of Hematology
PY - 2019/6/20
Y1 - 2019/6/20
N2 - Weibel-Palade bodies (WPBs) are secretory granules that contain von Willebrand factor and P-selectin, molecules that regulate hemostasis and inflammation, respectively. The presence of CD63/LAMP3 in the limiting membrane of WPBs has led to their classification as lysosome-related organelles. Many lysosome-related organelles contain intraluminal vesicles (ILVs) enriched in CD63 that are secreted into the extracellular environment during cell activation to mediate intercellular communication. To date, there are no reports that WPBs contain or release ILVs. By light microscopy and live-cell imaging, we show that CD63 is enriched in microdomains within WPBs. Extracellular antibody recycling studies showed that CD63 in WPB microdomains can originate from the plasma membrane. By cryo-electron tomography of frozen-hydrated endothelial cells, we identify internal vesicles as novel structural features of the WPB lumen. By live-cell fluorescence microscopy, we directly observe the exocytotic release of EGFP-CD63 ILVs as discrete particles from individual WPBs. WPB exocytosis provides a novel route for release of ILVs during endothelial cell stimulation.
AB - Weibel-Palade bodies (WPBs) are secretory granules that contain von Willebrand factor and P-selectin, molecules that regulate hemostasis and inflammation, respectively. The presence of CD63/LAMP3 in the limiting membrane of WPBs has led to their classification as lysosome-related organelles. Many lysosome-related organelles contain intraluminal vesicles (ILVs) enriched in CD63 that are secreted into the extracellular environment during cell activation to mediate intercellular communication. To date, there are no reports that WPBs contain or release ILVs. By light microscopy and live-cell imaging, we show that CD63 is enriched in microdomains within WPBs. Extracellular antibody recycling studies showed that CD63 in WPB microdomains can originate from the plasma membrane. By cryo-electron tomography of frozen-hydrated endothelial cells, we identify internal vesicles as novel structural features of the WPB lumen. By live-cell fluorescence microscopy, we directly observe the exocytotic release of EGFP-CD63 ILVs as discrete particles from individual WPBs. WPB exocytosis provides a novel route for release of ILVs during endothelial cell stimulation.
UR - http://www.scopus.com/inward/record.url?scp=85068367432&partnerID=8YFLogxK
U2 - 10.1182/blood-2018-09-874552
DO - 10.1182/blood-2018-09-874552
M3 - Article
C2 - 30760452
AN - SCOPUS:85068367432
SN - 0006-4971
VL - 133
SP - 2707
EP - 2717
JO - Blood
JF - Blood
IS - 25
ER -