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Status |
Public on Aug 26, 2014 |
Title |
Molecular and cellular characterization of Cord Blood derived, IDO expressing human fibrocystic Myeloid Derived Suppressor Cells |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by array
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Summary |
By restraining T cell activation and promoting regulatory T cell (Treg) expansion, myeloid-derived suppressor cells (MDSC) and tolerogenic dendritic cells (DC) (tDC) can control self-reactive and anti-graft effector T cells in autoimmunity and transplantation. Their therapeutic use and characterization, however, is limited by their scarce availability in the peripheral blood of tumor-free donors. In the present study we describe and characterize a novel population of myeloid suppressor cells, named fibrocytic MDSC (f-MDSC), that are differentiated from umbilical cord blood (UCB) precursors by a 4 day culture with FDA approved cytokines (rh-GM-CSF and rh-G-CSF). This MDSC subset, characterized by the expression of MDSC-, DC-, and fybrocyte-s associated markers, promotes Treg regulatory cells expansion and promote normoglycemia in a xenogeneic model of Type 1 Diabetes (T1D). In order to exert their pro-tolerogenic function, fibrocytic MDSC require direct contact with activated T cells, which leads to the induction and secretion of indoleamine 2,3 deoxygenase (IDO). This new myeloid subset may represent an important tool for the in vitro and in vivo production of Treg for the treatment of autoimmune diseases, and either prevention or control of allograft rejection.
Keywords: expression profiling by array
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Overall design |
Human Umbilical cord blood (UCB) samples were cultured for 4 days with GM-CSF and G-CSF after Ficoll-Paque gradient separation and red cell lysis. CD33+/IL-4Rα+ cells were sterilely sorted and part stored in Trizol for RNA extraction (BEFORE CONTACT), and part co-cultured with autologous PHA activated CD3+ T cells either in cell to cell contact (CONTACT) or using a transwell device to separate the two popyulations (TRANSWELL). Three days later, T cells were removed from the co-culture either by repeated washes with PBS or by simply removing the transwell device. Adherent cells were lysed with Trizol and stored at -80’C. RNA extraction was carry on with miRNAeasy mini kit from Quiagen; RNA QC was assessed by Agilent BioAnalyzer 6000 and samples were analyzed with Affymetrix Human 2.0 ST array
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Contributor(s) |
Zoso A, Mazza E, Bicciato S, Mandruzzato S, Bronte V, Serafini P, Inverardi L |
Citation(s) |
25113564, 26484135 |
Submission date |
Nov 14, 2013 |
Last update date |
Jul 29, 2019 |
Contact name |
Silvio Bicciato |
E-mail(s) |
silvio.bicciato@unipd.it
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Phone |
+39-049-827-6108
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Organization name |
University of Padova
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Department |
Molecular Medicine
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Street address |
via U. Bassi 59/b
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City |
Padova |
ZIP/Postal code |
35131 |
Country |
Italy |
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Platforms (1) |
GPL17930 |
[HuGene-2_0-st] Affymetrix Human Gene 2.0 ST Array [HuGene20stv1_Hs_ENTREZG_17.0.0] |
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Samples (23)
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GSM1264249 |
f-MDSC, before contact, replicate 4 |
GSM1264250 |
f-MDSC, before contact, replicate 5 |
GSM1264251 |
f-MDSC, before contact, replicate 6 |
GSM1264252 |
f-MDSC, before contact, replicate 7 |
GSM1264253 |
f-MDSC, contact, replicate 1 |
GSM1264254 |
f-MDSC, contact, replicate 2 |
GSM1264255 |
f-MDSC, contact, replicate 3 |
GSM1264256 |
f-MDSC, contact, replicate 4 |
GSM1264257 |
f-MDSC, contact, replicate 5 |
GSM1264258 |
f-MDSC, contact, replicate 6 |
GSM1264259 |
f-MDSC, contact, replicate 7 |
GSM1264260 |
HEK-293 cell line, replicate 1 |
GSM1264261 |
HEK-293 cell line, replicate 2 |
GSM1264262 |
f-MDSC, transwell replicate 1 |
GSM1264263 |
f-MDSC, transwell replicate 2 |
GSM1264264 |
f-MDSC, transwell replicate 3 |
GSM1264265 |
f-MDSC, transwell replicate 4 |
GSM1264266 |
f-MDSC, transwell replicate 5 |
GSM1264267 |
f-MDSC, transwell replicate 6 |
GSM1264268 |
f-MDSC, transwell replicate 7 |
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Relations |
BioProject |
PRJNA227735 |
Supplementary file |
Size |
Download |
File type/resource |
GSE52376_RAW.tar |
209.0 Mb |
(http)(custom) |
TAR (of CEL) |
Processed data included within Sample table |
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