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A microRNA profile of human CD8(+) regulatory T cells and characterization of the effects of microRNAs on Treg cell-associated genes.

Jebbawi F, Fayyad-Kazan H, Merimi M, Lewalle P, Verougstraete JC, Leo O, Romero P, Burny A, Badran B, Martiat P, Rouas R - J Transl Med (2014)

Bottom Line: We used the 'TargetScan' and 'miRBase' bioinformatics programs to identify potential target sites for these microRNAs in the 3'-UTR of important Treg cell-associated genes.We are examining the biological relevance of this 'signature' by studying its impact on other important Treg cell-associated genes.These efforts could result in a better understanding of the regulation of Treg cell function and might reveal new targets for immunotherapy in immune disorders and cancer.

View Article: PubMed Central - PubMed

Affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. Fadi.Jebbawi@ulb.ac.be.

ABSTRACT

Background: Recently, regulatory T (Treg) cells have gained interest in the fields of immunopathology, transplantation and oncoimmunology. Here, we investigated the microRNA expression profile of human natural CD8(+)CD25(+) Treg cells and the impact of microRNAs on molecules associated with immune regulation.

Methods: We purified human natural CD8(+) Treg cells and assessed the expression of FOXP3 and CTLA-4 by flow cytometry. We have also tested the ex vivo suppressive capacity of these cells in mixed leukocyte reactions. Using TaqMan low-density arrays and microRNA qPCR for validation, we could identify a microRNA 'signature' for CD8(+)CD25(+)FOXP3(+)CTLA-4(+) natural Treg cells. We used the 'TargetScan' and 'miRBase' bioinformatics programs to identify potential target sites for these microRNAs in the 3'-UTR of important Treg cell-associated genes.

Results: The human CD8(+)CD25(+) natural Treg cell microRNA signature includes 10 differentially expressed microRNAs. We demonstrated an impact of this signature on Treg cell biology by showing specific regulation of FOXP3, CTLA-4 and GARP gene expression by microRNA using site-directed mutagenesis and a dual-luciferase reporter assay. Furthermore, we used microRNA transduction experiments to demonstrate that these microRNAs impacted their target genes in human primary Treg cells ex vivo.

Conclusions: We are examining the biological relevance of this 'signature' by studying its impact on other important Treg cell-associated genes. These efforts could result in a better understanding of the regulation of Treg cell function and might reveal new targets for immunotherapy in immune disorders and cancer.

No MeSH data available.


Related in: MedlinePlus

Differential expression of miR-24, −335, −155, −31, −210, −449, −509, −214, −205 and −9 between CD8+CD25+nTregs and CD8+CD25−T cells. Data obtained by qRT-PCR amplification of miRs are plotted. p values for each miRNA relative expression are presented. Boxes represent SE; and Error bars SD Pooled data from five independent experiments are shown. (*p < 0.05; **p < 0.01 CD8+CD25+ nTregs versus CD8+CD25− T cells (Student's t test).
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Fig3: Differential expression of miR-24, −335, −155, −31, −210, −449, −509, −214, −205 and −9 between CD8+CD25+nTregs and CD8+CD25−T cells. Data obtained by qRT-PCR amplification of miRs are plotted. p values for each miRNA relative expression are presented. Boxes represent SE; and Error bars SD Pooled data from five independent experiments are shown. (*p < 0.05; **p < 0.01 CD8+CD25+ nTregs versus CD8+CD25− T cells (Student's t test).

Mentions: We first studied miRNAs from five independent CD8+CD25+ natural Treg cell and CD8+CD25− T cell samples using the TLDA technique. We could identify several miRNAs that were differentially expressed between CD8+CD25+ natural Treg cells and CD8+CD25− T cells. Differentially expressed miRNAs were validated by real-time PCR (Figure 3). A Treg cell miRNA signature was identified that included 10 significantly differentially expressed miRNAs: miR-9, −24, −31, −155, −210, −335 and −449 were downregulated in CD8+ natural Treg cells, whereas miR-214, −205 and −509 were overexpressed.Figure 3


A microRNA profile of human CD8(+) regulatory T cells and characterization of the effects of microRNAs on Treg cell-associated genes.

Jebbawi F, Fayyad-Kazan H, Merimi M, Lewalle P, Verougstraete JC, Leo O, Romero P, Burny A, Badran B, Martiat P, Rouas R - J Transl Med (2014)

Differential expression of miR-24, −335, −155, −31, −210, −449, −509, −214, −205 and −9 between CD8+CD25+nTregs and CD8+CD25−T cells. Data obtained by qRT-PCR amplification of miRs are plotted. p values for each miRNA relative expression are presented. Boxes represent SE; and Error bars SD Pooled data from five independent experiments are shown. (*p < 0.05; **p < 0.01 CD8+CD25+ nTregs versus CD8+CD25− T cells (Student's t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4440568&req=5

Fig3: Differential expression of miR-24, −335, −155, −31, −210, −449, −509, −214, −205 and −9 between CD8+CD25+nTregs and CD8+CD25−T cells. Data obtained by qRT-PCR amplification of miRs are plotted. p values for each miRNA relative expression are presented. Boxes represent SE; and Error bars SD Pooled data from five independent experiments are shown. (*p < 0.05; **p < 0.01 CD8+CD25+ nTregs versus CD8+CD25− T cells (Student's t test).
Mentions: We first studied miRNAs from five independent CD8+CD25+ natural Treg cell and CD8+CD25− T cell samples using the TLDA technique. We could identify several miRNAs that were differentially expressed between CD8+CD25+ natural Treg cells and CD8+CD25− T cells. Differentially expressed miRNAs were validated by real-time PCR (Figure 3). A Treg cell miRNA signature was identified that included 10 significantly differentially expressed miRNAs: miR-9, −24, −31, −155, −210, −335 and −449 were downregulated in CD8+ natural Treg cells, whereas miR-214, −205 and −509 were overexpressed.Figure 3

Bottom Line: We used the 'TargetScan' and 'miRBase' bioinformatics programs to identify potential target sites for these microRNAs in the 3'-UTR of important Treg cell-associated genes.We are examining the biological relevance of this 'signature' by studying its impact on other important Treg cell-associated genes.These efforts could result in a better understanding of the regulation of Treg cell function and might reveal new targets for immunotherapy in immune disorders and cancer.

View Article: PubMed Central - PubMed

Affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. Fadi.Jebbawi@ulb.ac.be.

ABSTRACT

Background: Recently, regulatory T (Treg) cells have gained interest in the fields of immunopathology, transplantation and oncoimmunology. Here, we investigated the microRNA expression profile of human natural CD8(+)CD25(+) Treg cells and the impact of microRNAs on molecules associated with immune regulation.

Methods: We purified human natural CD8(+) Treg cells and assessed the expression of FOXP3 and CTLA-4 by flow cytometry. We have also tested the ex vivo suppressive capacity of these cells in mixed leukocyte reactions. Using TaqMan low-density arrays and microRNA qPCR for validation, we could identify a microRNA 'signature' for CD8(+)CD25(+)FOXP3(+)CTLA-4(+) natural Treg cells. We used the 'TargetScan' and 'miRBase' bioinformatics programs to identify potential target sites for these microRNAs in the 3'-UTR of important Treg cell-associated genes.

Results: The human CD8(+)CD25(+) natural Treg cell microRNA signature includes 10 differentially expressed microRNAs. We demonstrated an impact of this signature on Treg cell biology by showing specific regulation of FOXP3, CTLA-4 and GARP gene expression by microRNA using site-directed mutagenesis and a dual-luciferase reporter assay. Furthermore, we used microRNA transduction experiments to demonstrate that these microRNAs impacted their target genes in human primary Treg cells ex vivo.

Conclusions: We are examining the biological relevance of this 'signature' by studying its impact on other important Treg cell-associated genes. These efforts could result in a better understanding of the regulation of Treg cell function and might reveal new targets for immunotherapy in immune disorders and cancer.

No MeSH data available.


Related in: MedlinePlus