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CITE-seq of in vitro control and memory-like human NK cells

ABSTRACT: CITE-seq of in vitro control and memory-like human NK cells

PROVIDER: PRJNA771661 | ENA |

REPOSITORIES: ENA

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CITE-seq of in vitro control and memory-like human NK cells

Project description:Peripheral blood NK cells were cultured into either control or cytokine-induced memory-like NK cells as described in PMID: 32614951. After differentiation, cells were labeled with hashtag antibodies and cite-seq antibodies and control and memory-like NK cells were combined and loaded onto the 10x Genomics Chromium Instrument a the McDonnell Genome Institute at Washington University School of Medicine. The following samples were analyzed to develop a classification tool for unbiased identification of memory-like NK cells in a bulk single-cell RNA-seq population.

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Project description:Here we show that Zika virus infection induces memory like NK cells that express CD27. This subset shared transcriptional changes with memory CD8 T cells, stem cells and stem like T cells. These NK cells with memory and stemness features which we term NK memory stem cells (SCMNK) demonstrated greater antiviral potential. Transcriptional data of SCMNK and non-memory NK cells is presented.

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Project description:NK cells are an emerging cancer cellular therapy and potent mediators of anti-tumor immunity. Cytokine-induced memory-like (ML) NK cellular therapy is safe and induces remissions in acute myeloid leukemia (AML) patients. However, the dynamic molecular changes that occur after memory-like differentiation in vitro are unclear. Here, control or ML NK cells purified from normal donor PBMC were generated in vitro. Briefly, RosetteSep-purified NK cells were incubated in IL-12, IL-15, and IL-18, or low-dose IL-15 as a control for 16-18 hours. Control or cytokine-activated NK cells were washed three times and cultured for 6 days in low-dose IL-15, which is required for NK cell survival. After 6 days, RNA was isolated from control and memory-like (ML) NK cells (IL12/15/18 activation) and RNA-sequencing performed. Because the transcription factor GATA-3 was increased specifically in ML NK cells, we hypothesized ML NK cells would exhibit a GATA-3 gene signature compared to control NK cells. Indeed, using GSEA, a significant gene signature was associated with ML NK cell differentiation. These data support the role for GATA-3 in regulating the ML NK cell molecular program.

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NK cell response to IL2 in vitro

Project description:Here we analyzed the transcriptional response to IL2 in NK cells in vitro. Splenic NK cells were isolated by negative selection from 6wk old male C57BL/6 mice. These were directly cultured with IL2 (250U/ml) for 8hrs in vitro. Cells were then transferred directly into TriZol. RNA was prepared in Trizol for gene expression profiling by Affymetrix Mouse Gene 1.0 ST Arrays. The samples in this series are re-analyzed samples from GSE84562.

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Project description:In this study, we show that CD27+ memory like NK cells generated following Zika virus infection exhibited stem-like features viz., self-renewal pathway, differentiation into effector cells and longer telomeres, and greater therapeutic potential than CD27- and naive CD27+ NK cells when adoptively transferred to Zika virus infected mice. In addition, epigenetic landscape of CD27+ memory like NK is markedly different compared to CD27- NK cells.

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