Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.

Exosomes are membrane-bound vesicles found in all biological fluids. AML patients' plasma collected at diagnosis contains elevated exosome levels relative to normal donor (ND) plasma. The molecular profile of AML exosomes changes in the course of therapy and may serve as a measure of disease pr...

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Autores principales:	Chang Sook Hong, Laurent Muller, Michael Boyiadzis, Theresa L Whiteside
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/57dc9061dc854703ac25174633bdcea9
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spelling	oai:doaj.org-article:57dc9061dc854703ac25174633bdcea92021-11-25T06:06:03ZIsolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.1932-620310.1371/journal.pone.0103310https://doaj.org/article/57dc9061dc854703ac25174633bdcea92014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25093329/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Exosomes are membrane-bound vesicles found in all biological fluids. AML patients' plasma collected at diagnosis contains elevated exosome levels relative to normal donor (ND) plasma. The molecular profile of AML exosomes changes in the course of therapy and may serve as a measure of disease progression or response to therapy. However, plasma contains a mix of exosomes derived from various cell types. To be able to utilize blast-derived exosomes as biomarkers for AML, we have developed an immunoaffinity-based capture method utilizing magnetic microbeads coated with anti-CD34 antibody (Ab). This Ab is specific for CD34, a unique marker of AML blasts. The capture procedure was developed using CD34+ exosomes derived from Kasumi-1 AML cell culture supernatants. The capture capacity of CD34microbeads was shown to linearly correlate with the input exosomes. A 10 uL aliquot of CD34 microbeads was able to capture all of CD34+ exosomes present in 100-1,000 uL of AML plasma. The levels of immunocaptured CD34+ exosomes correlated with the percentages of CD34+ blasts in the AML patients' peripheral blood. The immunocaptured exosomes had a typical cup-shaped morphology by transmission electron microscopy, and their molecular cargo was similar to that of parental blasts. These exosomes were biologically-active. Upon co-incubation with natural killer (NK) cells, captured blast-derived exosomes down-regulated surface NKG2D expression, while non-captured exosomes reduced expression levels of NKp46. Our data provide a proof-of-principle that blast-derived exosomes can be quantitatively recovered from AML patients' plasma, their molecular profile recapitulates that of autologous blasts and they retain the ability to mediate immune suppression. These data suggest that immunocaptured blast-derived exosomes might be useful in diagnosis and/or prognosis of AML in the future.Chang Sook HongLaurent MullerMichael BoyiadzisTheresa L WhitesidePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 8, p e103310 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Chang Sook Hong Laurent Muller Michael Boyiadzis Theresa L Whiteside Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
description	Exosomes are membrane-bound vesicles found in all biological fluids. AML patients' plasma collected at diagnosis contains elevated exosome levels relative to normal donor (ND) plasma. The molecular profile of AML exosomes changes in the course of therapy and may serve as a measure of disease progression or response to therapy. However, plasma contains a mix of exosomes derived from various cell types. To be able to utilize blast-derived exosomes as biomarkers for AML, we have developed an immunoaffinity-based capture method utilizing magnetic microbeads coated with anti-CD34 antibody (Ab). This Ab is specific for CD34, a unique marker of AML blasts. The capture procedure was developed using CD34+ exosomes derived from Kasumi-1 AML cell culture supernatants. The capture capacity of CD34microbeads was shown to linearly correlate with the input exosomes. A 10 uL aliquot of CD34 microbeads was able to capture all of CD34+ exosomes present in 100-1,000 uL of AML plasma. The levels of immunocaptured CD34+ exosomes correlated with the percentages of CD34+ blasts in the AML patients' peripheral blood. The immunocaptured exosomes had a typical cup-shaped morphology by transmission electron microscopy, and their molecular cargo was similar to that of parental blasts. These exosomes were biologically-active. Upon co-incubation with natural killer (NK) cells, captured blast-derived exosomes down-regulated surface NKG2D expression, while non-captured exosomes reduced expression levels of NKp46. Our data provide a proof-of-principle that blast-derived exosomes can be quantitatively recovered from AML patients' plasma, their molecular profile recapitulates that of autologous blasts and they retain the ability to mediate immune suppression. These data suggest that immunocaptured blast-derived exosomes might be useful in diagnosis and/or prognosis of AML in the future.
format	article
author	Chang Sook Hong Laurent Muller Michael Boyiadzis Theresa L Whiteside
author_facet	Chang Sook Hong Laurent Muller Michael Boyiadzis Theresa L Whiteside
author_sort	Chang Sook Hong
title	Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
title_short	Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
title_full	Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
title_fullStr	Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
title_full_unstemmed	Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.
title_sort	isolation and characterization of cd34+ blast-derived exosomes in acute myeloid leukemia.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/57dc9061dc854703ac25174633bdcea9
work_keys_str_mv	AT changsookhong isolationandcharacterizationofcd34blastderivedexosomesinacutemyeloidleukemia AT laurentmuller isolationandcharacterizationofcd34blastderivedexosomesinacutemyeloidleukemia AT michaelboyiadzis isolationandcharacterizationofcd34blastderivedexosomesinacutemyeloidleukemia AT theresalwhiteside isolationandcharacterizationofcd34blastderivedexosomesinacutemyeloidleukemia
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Isolation and characterization of CD34+ blast-derived exosomes in acute myeloid leukemia.

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