HUMAN HEMATOPOIETIC STEM CELLS GENERATE EXOSOMES CONTAINING ACTIVE PROTEASOMES BUT NOT CASPASES
I.M. Prudnikov, V.M. Tsyvkin, A.M. Smirnov, I.V. Pristash
O.O. Bogomoletsz Institute of Physiology of NAS of Ukraine, Kyiv, Ukraine
DOI: https://doi.org/10.15407/fz66.06.013
Abstract
Activities of two classes of neutral proteases were studied:
caspases and proteasomes, which are contained in extracellular
vesicles generated by mesenchymal and hematopoietic human
stem cells (MSCs and HSCs, respectively). The formation
of apoptosomes induced by the cytochrome C and dATP
did not occurred in these cells. The presence of TNFa in the
culture medium of HSCs causes the appearance of caspase-3
intracellular activity. Herewith caspase activity was detected
in exosomes also. This activity was completely inhibited by
a non-substrate caspase inhibitor, emricasane, and it was
not sensitive to proteasome inhibitors. It is assumed that the
caspases’ activity from the cultural medium is the sum of
membrane (outer side of exosome membrane), intracellular
and extracellular activities. Apparently, this activity refers to
apoptosomes as it was revealed by gel filtration of cultural
medium with TNFa free from exosomes. Thus, TNFa induces
the appearance of neutrophils in the culture of HSCs or the
generation of other differentiated cells that are capable of
apoptosis, in contrast to HSCs or MSCs. Proteasomal activity
increased noticeably in exosomes from HSCs after the addition
of TNFa. In contrast to exosomes from MSCs, all activity of
proteasomes in HSCs-produced exosomes had a membrane
localization.
Keywords:
apoptosis; proteasomes; human stem cells; exosomes
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