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HLADR expression marks recently divided memory CD4 T cells in latent tuberculosis infected subjects upon in vitro stimulation with Mycobacterium tuberculosis antigens

ABSTRACT: HLADR expression marks recently divided memory CD4 T cells in latent tuberculosis infected subjects upon in vitro stimulation with Mycobacterium tuberculosis antigens

PROVIDER: PRJNA682752 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13202400_1.fastq.gz	Fastqsanger.gz
	SRR13202400_2.fastq.gz	Fastqsanger.gz
	SRR13202401_1.fastq.gz	Fastqsanger.gz
	SRR13202401_2.fastq.gz	Fastqsanger.gz
	SRR13202402_1.fastq.gz	Fastqsanger.gz

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HLADR expression marks recently divided memory CD4 T cells in latent tuberculosis infected subjects upon in vitro stimulation with Mycobacterium tuberculosis antigens

Project description:Upon antigen encounter, T cells can rapidly divide and form short-lived effector cells which plays an important role fighting an active infection. However, little is known about the molecular markers that distinguish such effector cells from other T cell populations in humans. Due to previous exposure with Mycobacterium tuberculosis (Mtb), we expect latent tuberculosis infected (LTBI) subjects to have Mtb-specific memory CD4 T cells that are capable of rapid expansion upon antigen re-encounter. To investigate this further we designed an in vitro MTP-AE (Memory T cell Proliferation upon Antigen Exposure) assay to track and isolate proliferated CD4 T cells in response to antigen stimulation. PBMCs from LTBI subjects were cultured with the MTB300 (Mtb specific peptide pool, PMID: 27409590) for 8 days before sorting for HLADR+ vs HLADR- and divided vs undivided CD4 T cells. We found that HLADR is a marker of recently divided memory CD4 T cells and that differentially expessed genes in HLADR+/recently divided cells were enriched for cytotoxic markers and cytokines compared to HLADR-/undivided cells.

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Project description:For further identify the differentiation between latent and clinical tuberculosis (TB), we employed whole genome microarray expression profiling to study genes with significant expression change in peripheral CD4+T cells between healthy control, latent tuberculosis (LTB) and clinical tuberculosis (TB). Our experiment included 4 groups: healthy donor (HD), latent TB1 (LTB1) with low IFN-gamma release level, latent TB2 (LTB2) with high IFN-gamma release level, and tuberculosis (TB) with high IFN-gamma release level. Human peripheral blood mononuclear cells were collected, from which CD4+T cells were isolated. Total RNA of each individuals of each group was extracted from peripheral CD4+T cells. One μg of RNA mixture, pooled equivalently by each individual total RNA of each group, was administrated microarray test. Compared with HD, through analyzing enriched-Gene Ontology (GO) terms and KEGG pathways of each group, we found peripheral CD4+T cells might had different ability for mycobacterium tuberculosis infection in LTB1, LTB2 and TB. Finally we detected that TNFSF13/APRIL and TNFSF13B/BAFF was significant up-regulation in both CD4+T cells and serum of TB by real time PCR and ELISA, respectively.

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