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Gene expression signature of ex vivo expanded NK(CD3-CD56+) cells for 14 days

ABSTRACT: Gene expression signature of ex vivo expanded NK(CD3-CD56+) cells for 14 days

PROVIDER: PRJNA686645 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13275812_1.fastq.gz	Fastqsanger.gz
	SRR13275812_2.fastq.gz	Fastqsanger.gz
	SRR13275813_1.fastq.gz	Fastqsanger.gz
	SRR13275813_2.fastq.gz	Fastqsanger.gz
	SRR13275814_1.fastq.gz	Fastqsanger.gz

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Gene expression signature of ex vivo expanded NK(CD3-CD56+) cells for 14 days

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Gene expression analysis for CD56- T, NK, CD56+ T cells, and iNKT cells

Project description:A small subset of T cells also expresses kiler-cell immunoglobulin-like receptors (KIRs). We find that KIR+ T cells primarily reside in the CD56+ T population. However, little is known on how these cells are different from the conventional CD56- T, NK, and iNKT cells. We used microarray profiling to compare and determine the distinctive differences of CD56+ T cell and its KIR subsets when compared to the conventional CD56- T, NK and iNKT cells. Lymphocyte subsets were sorted from human peripheral blood mononuclear cells with FACSAriaII (BD Biosciences, San Jose, CA) using anti-CD3, anti-CD56, anti-CD14, anti-KIR2DL1, anti-KIR2DL2/3, anti-KIR3DL1 and anti-TCRValpha24 antibodies. The purity of CD3+CD56- T cells, CD3-CD56+ NK cells, CD3+CD56+ T cells, KIR-CD3+CD56+ T cells, and KIR+CD3+CD56+ T cells were more than 98% in all experiments. The purities of iNKT cells for TCRValpha24 and CD1d-tetramer were >95% and >90%, respectively. RNA pre-amplification, labeling and hybridization on Human Genome U133Plus 2.0 GeneChip array were performed in the St. Jude Hartwell Center for Bioinformatics & Biotechnology microarray core facility according to the manufacturer’s instructions (Affymetrix, Santa Clara, CA).

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Project description:we performed RNA sequencing to employ an unbiased approach to identify the altered gene expression enhancing the function of ex vivo expanded pNK cells for 14 days, and transcriptionally compared day 0 and day 14 pNK cells.

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CAR-NK cells derived from cord blood predominantly originated from CD56-CD7+CD34-HLA-DR- lineage markers- NK progenitor cells

Project description:Cord blood (CB)-derived chimeric antigen receptor (CAR)-natural killer (NK) cells targeting CD19 has been shown to be effective against B cell malignancies. While human CD56+ NK cells can be expanded in vitro, it is also known that NK cells can be differentiated from hematopoietic progenitor cells. It is still unclear whether CAR-NK cells are originated from mature NK cells or NK progenitor cells in CB. Here we found that CAR-NK cells are predominantly derived from the CD56- NK progenitor cells. We first found that substantial numbers of CD19 CAR-NK cells were produced from the CD56- CB mononuclear cells after in vitro culture for two weeks. Single cell RNAseq analysis of CD56-CD3-CD14-CD19- CB mononuclear cells revealed that those cells were subdivided into three subpopulations based on the expression of CD34 and HLA-DR. NK cells were predominantly produced from the CD34-HLA-DR- cells. In addition, in the CD34-HLA-DR- cells, only CD7+ cells could differentiate into NK cells. These results indicate that CD56-CD7+CD34- HLA-DR-lineage marker (Lin)- cells are the major origin of human CB-derived CAR-NK cells, indicating that we need to develop methods to enhance the quality and quantity of NK cells produced from these NK cell progenitor cells.

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Gene expression signature of ex vivo expanded NK cells for 14 days

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