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P14 CD8 T cells during acute LCMV infection

ABSTRACT: P14 CD8 T cells during acute LCMV infection

PROVIDER: PRJNA632045 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11781400.fastq.gz	Fastqsanger.gz
	SRR11781401.fastq.gz	Fastqsanger.gz
	SRR11781402.fastq.gz	Fastqsanger.gz
	SRR11781403.fastq.gz	Fastqsanger.gz
	SRR11781404.fastq.gz	Fastqsanger.gz

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RNA-seq of P14 CD8 T cells during acute LCMV infection

Project description:We performed RNA-seq, ATAC-seq, and H3K27Ac ChIP-seq on P14 CD8 T cells during acute infection with LCMV Armstrong on days 0 (naïve cells), 9 (memory precursor effector cells (MPEC) and short-lived effector cells (SLECs)), and 40+ (memory cells) post infection.

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Project description:Understanding the response of memory CD8 T cells to persistent antigen re-stimulation and the role of CD4 T cell help is critical to the design of successful vaccines for chronic diseases. However, studies comparing the protective abilities and qualities of memory and naïve cells have been mostly performed in acute infections, and little is known about their roles during chronic infections. Herein, we show that memory cells dominate over naïve cells and are protective when present in large enough numbers to quickly reduce infection. In contrast, when infection is not rapidly reduced, memory cells are quickly lost, unlike naïve cells. This loss of memory cells is due to (i) an early block in cell proliferation, (ii) selective regulation by the inhibitory receptor 2B4, and (iii) increased reliance on CD4 T cell help. These findings have important implications towards the design of T cell vaccines against chronic infections and tumors. 16 samples are analyzed: 3 replicates of secondary effector CD8 P14 T cells at day 8 post-acute lymphocytic choriomeningitis virus (LCMV) infection; 4 replicates of secondary effector CD8 P14 T cells at day 8 post-chronic LCMV infection; 4 replicates of primary effector CD8 P14 T cells at day 8 post-acute LCMV infection; and 5 replicates of primary effector CD8 P14 T cells at day 8 post-chronic LCMV infection.

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Project description:The PI3K/Akt signaling pathway impacts various aspects of CD8 T cell homeostasis, such as effect versus memory cell differentiation, during viral infection. We used microarrays to determine which downstream molecules were affected and what other signaling pathways were interconnected with the Akt pathway by constitutive activation of Akt in LCMV-infected CD8 T cells. Splenocytes from naive P14/WT or P14/Akt mice were stained with anti-CD8 and anti-Ly5.1, and CD8 T cells were sorted using a FACSAria II instrument. Purified Ly5.1+ CD8 T cells from P14/WT or P14/Akt mice were transferred into B6 mice, which were subsequently infected with LCMV Armstrong. At day 8 post infection, splenocytes were stained with anti-CD8, anti-Ly5.1, anti-KLRG1, and anti-CD127. Following staining, short-lived effector cells (SLECs) and memory precursor effector cells (MPECs) were sorted using the FACSAria II instrument; the purity of the sorted cells was >95%. A total of 5 samples were analyzed, including WT naive, WT SLEC, WT MPEC, Akt naive and Akt SLEC.

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H3K27Ac ChIP-seq of P14 CD8 T cells during acute LCMV infection

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