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SRX8472224: GSM4591271: R294: CFSE high CD69 pos; Erythrocebus patas; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2500) run: 34.1M spots, 3.4G bases, 1.9Gb downloads

Submitted by: NCBI (GEO)
Study: SIVagm coevolutions lead to altered epigenetic control of CD4 expression [RNA-seq]
show Abstracthide Abstract
African green monkeys (AGMs) are natural hosts of Simian immunodeficiency virus (SIV) that post-thymically down-regulate CD4 to maintain a large population of CD4-CD8aa+ virus-resistant cells with T-helper functionality. AGMs can become aviremic and seemingly cured of SIV by down-regulating CD4 in all CD4+ T cells in vivo. To understand the mechanisms of this process we performed genome-wide transcriptional analysis on T cells induced to down-regulate CD4 in vitro from AGMs and closely-related Patas monkeys, and T cells that maintain CD4 expression from rhesus macaques. In cells that down-regulated CD4, pathway analysis of genes common to AGM and Patas natural hosts yet unique from divided rhesus CD4+ T cells revealed an atypical regulation of the DNA methylation machinery, which de-repressed CD4 expression when pharmacologically targeted with 5-aza-2 deoxycytidine. This signature was driven largely by the dioxygenase TET3 that became down-regulated with loss of CD4 expression. CpG motifs within the AGM CD4 promoter region became methylated during CD4 downregulation in vitro and were stably imprinted in AGM CD4-CD8aa+ T cells sorted directly ex vivo. These results suggest AGMs employ epigenetic mechanisms to durably silence the CD4 gene. Manipulation of these mechanisms could provide avenues for modulating SIV and human immunodeficiency virus (HIV)-1 entry receptor expression in hosts that become progressively SIV-infected, which could lead to novel therapeutic interventions aimed to reduce HIV viremia in vivo. Overall design: Examination of genes uniquely regulated in natural host CD4 T cells induced to down-regulate CD4 in vitro when compared to progressive host CD4 T cells that maintain CD4 expression upon in vitro stimulation. Cells were sorted on the basis of divided or non-divided (ie CFSE high/low), CD69 expression and CD4 expression after 5 days of stimulation with SEB. Upon sample processing, all samples were assigned the abbreviation "JB" (for Jason Brenchley) and randomized 1-54. The abbreviation "AG" in title column specifies all samples from African green monkeys and number specifies animal ID. All samples beginning with "R" are for Patas monkeys. All other animal ID's specify samples from rhesus.
Sample: R294: CFSE high CD69 pos
SAMN15102158 • SRS6772837 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was isolated following the Qiagen RNeasy Micro Kit protocol. Samples were purified through Qiagen columns with on-column DNase treatment for 15 minutes . Libraries were made from purified total RNA using the Illumina TruSeq Stranded Total RNA Sample Preparation Kit protocol. rRNA was removed from samples by denaturing and then purified on rRNA removal beads. RNA was then fragmented and primed with random hexamers for first-strand synthesis in a single step on a thermocycler for 8 minutes at 94°C. First-strand synthesis immediately followed using Superscript II polymerase (Invitrogen) on a thermocycler with the following parameters: 25°C for 10mins, 42°C for 15 mins and 70°C 15 mins. In order to determine which strand the transcript was transcribed from, dUTP's were used instead of dTTP's during the second strand synthesis. Single adenines were added to the blunted ds cDNA. Illumina adapters containing unique dual-indices were ligated to each library. This allows the samples to be multiplexed on the sequencer. Libraries containing Illumina adapters were enriched using 15-18 cycles of PCR. Libraries were sequenced on an Illumina HiSeq 2500 (Illumina. San Diego, CA).
Experiment attributes:
GEO Accession: GSM4591271
Links:
Runs: 1 run, 34.1M spots, 3.4G bases, 1.9Gb
Run# of Spots# of BasesSizePublished
SRR1192657834,052,0963.4G1.9Gb2020-10-07

ID:
11012989

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