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| Status |
Public on Jan 10, 2015 |
| Title |
microRNA expression profiling of different T cell subset in human tonsils |
| Organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
Tight control of follicular helper T(Tfh) cells is required for optimal maturation of the germinal center (GC) response. The molecular mechansims controlling Tfh cell differentiation remain incompletely understood. Here we sought to to identfiy miRNAs that might regulate Tfh cell development and/or function.
To identfiy miRNAs that are differentially expressed in Tfh cells, we performed Agilent microRNA microarray analysis comparing Tfh cells with the other main T cell subsets (naive T cells, non-Tfh effector or non-Tfh memory T cells, CD57+ Tfh cells and CD57- Tfh cells).
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| Overall design |
RNA was extracted from 4 separate human tonsils (biological replicates) using mirVANA RNA isolation kit (Ambion) according to the manufacturer’s protocol. Each biological sample was analysed individually. Live lymphocytes were sorted based on negative 7AAD staining, and thefollowing four subsets were purified: Naive T cells (CD4+ CD45RO-); CD57+ Tfh cells (CD4+ CD45RO+ PD-1_high CXCR5_high CD57+); CD57- Tfh cells (CD4+ CD45RO+ PD-1_high CXCR5_high CD57-) and non-Tfh effector or memory T cells (CD4+ CD45RO+ PD-1_neg CXCR5_neg). RNA quantity and quality was determined using a Nanodrop 1000 and an Agilent 2100 Bioanalyzer respectively. Hybridization on Agilent Human miRNA Microarray (V1) Kit, 8x15K and was performed at The Ramaciotti Center for Genomics (University of New South Wales, Australia). The mean of 3-4 biological replicates from each population of each population were calculated and values were plotted using Prism.
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| Contributor(s) |
Srivastava M, Pratama A, Rigby R, Vinuesa CG |
| Citation(s) |
25743066 |
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| Submission date |
Jan 09, 2015 |
| Last update date |
Oct 20, 2015 |
| Contact name |
Monika Srivastava |
| E-mail(s) |
[email protected]
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| Phone |
0061-2-61255056
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| Organization name |
JCSMR, ANU
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| Department |
Pathogen and Immunity
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| Street address |
Building 131, Garran Road
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| City |
Canberra |
| State/province |
ACT |
| ZIP/Postal code |
2601 |
| Country |
Australia |
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| Platforms (1) |
| GPL19622 |
Agilent-019118 Human miRNA Microarray V1 G4470A [miRBase release 9.1 miRNA ID version] |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA272140 |
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