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Critical role of extracellular vesicles in modulating the cellular effects of cytokines

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Abstract

Under physiological and pathological conditions, extracellular vesicles (EVs) are present in the extracellular compartment simultaneously with soluble mediators. We hypothesized that cytokine effects may be modulated by EVs, the recently recognized conveyors of intercellular messages. In order to test this hypothesis, human monocyte cells were incubated with CCRF acute lymphoblastic leukemia cell line-derived EVs with or without the addition of recombinant human TNF, and global gene expression changes were analyzed. EVs alone regulated the expression of numerous genes related to inflammation and signaling. In combination, the effects of EVs and TNF were additive, antagonistic, or independent. The differential effects of EVs and TNF or their simultaneous presence were also validated by Taqman assays and ELISA, and by testing different populations of purified EVs. In the case of the paramount chemokine IL-8, we were able to demonstrate a synergistic upregulation by purified EVs and TNF. Our data suggest that neglecting the modulating role of EVs on the effects of soluble mediators may skew experimental results. On the other hand, considering the combined effects of cytokines and EVs may prove therapeutically useful by targeting both compartments at the same time.

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Abbreviations

CNR2:

Cannabinoid receptor 2

EV:

Extracellular vesicle

Gex (HPRT):

Gene expression (relative to HPRT)

GSEA:

Gene set enrichment analysis

HPRT:

Hypoxanthine-guanine phosphoribosyltransferase

ICAM1:

Intracellular adhesion molecule 1

IL-8:

Interleukin 8

KEGG:

Kyoto Encyclopedia of Genes and Genomes

NPC1:

Niemann–Pick disease C1

MMP9:

Matrix metalloproteinase 9

SMPD3:

Sphingomyelin phosphodiesterase 3

TEM:

Transmission electron microscopy

TNF:

Tumor necrosis factor

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Acknowledgments

We are grateful for Ms. Andrea Orbán’s and Mrs. Rita Antónia Fekete Heszné’s excellent technical assistance and Eszter Tóth for her contribution to some of the experiments. This work was funded by OTKA NK 84043, OTKA PD104369, Baross Gábor (REG-KM-09-1-2009-0010), and Marie Curie Networks for Initial Training-ITN-FP7-PEOPLE-2011-ITN, PITN-GA-2011-289033. Furthermore, this research was supported by TÁMOP 4.2.4. A/1-11-1-2012-0001 “National Excellence Program––Elaborating and operating an inland student and researcher personal support system” and subsidized by the European Union and co-financed by the European Social Fund.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary

    Géza Tamás Szabó, Bettina Tarr, Krisztina Pálóczi, Katalin Éder, Eszter Lajkó, Sára Tóth, Bence György, Mária Pásztói, Andrea Németh, Xabier Osteikoetxea, Éva Pállinger, András Falus, Katalin Szabó-Taylor & Edit Irén Buzás

  2. Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary

    Ágnes Kittel

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  1. Géza Tamás Szabó
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Correspondence to Edit Irén Buzás.

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K. Szabó-Taylor and E. I. Buzás contributed equally.

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Szabó, G.T., Tarr, B., Pálóczi, K. et al. Critical role of extracellular vesicles in modulating the cellular effects of cytokines. Cell. Mol. Life Sci. 71, 4055–4067 (2014). https://doi.org/10.1007/s00018-014-1618-z

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