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2014 ; 2
(ä): 396-8
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Whole transcriptome analysis for T cell receptor-affinity and IRF4-regulated
clonal expansion of T cells
#MMPMID26484137
Shi W
; Man K
; Smyth GK
; Nutt SL
; Kallies A
Genom Data
2014[Dec]; 2
(ä): 396-8
PMID26484137
show ga
Clonal population expansion of T cells during an immune response is dependent on
the affinity of the T cell receptor (TCR) for its antigen [1]. However, there is
little understanding of how this process is controlled transcriptionally. We
found that the transcription factor IRF4 was induced in a manner dependent on
TCR-affinity and was critical for the clonal expansion and maintenance of
effector function of antigen-specific CD8(+) T cells. We performed a genome-wide
expression profiling experiment using RNA sequencing technology (RNA-seq) to
interrogate global expression changes when IRF4 was deleted in CD8(+) T cells
activated with either a low or high affinity peptide ligand. This allowed us not
only to determine IRF4-dependent transcriptional changes but also to identify
transcripts dependent on TCR-affinity [2]. Here we describe in detail the
analyses of the RNA-seq data, including quality control, read mapping,
quantification, normalization and assessment of differential gene expression. The
RNA-seq data can be accessed from Gene Expression Omnibus database (accession
number GSE49929).