Unknown,Transcriptomics,Genomics,Proteomics

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Expression profile of human natural killer cells co-cultured with P. falciparum infected erythrocytes

ABSTRACT: In order to identify an expression signature that might characterize the impact of Plasmodium parasites on NK cells, we have used Affymetrix oligonucleotide microarrays to examin the global gene expression profile of primary NK cells that have been co-incubated with P. falciparum infected red blood cells and have compared it to the expression pattern of NK genes induced by cytokine treatment. PBMCs were isolated from 3 healthy donors and were incubated with iRBCs, uRBCs, IL12 plus IL18 and without any stimulus. After 24 hours at 37ºC and 5%CO2, Natural killer cells were isolated by negative selection, checked for purity by flow cytometry and processed for RNA extraction and analysis with the Human Gene 1.0 Affymetrix array. The experiment was repeated 3 times (1-2 weeks apart) for each one of the 3 donors.

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Bonin

PROVIDER: E-GEOD-24791 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Natural Killer cells (NK), a major constituent of innate immune system, have the ability to kill the transformed and infected cells without prior sensitization; can be put to immunotherapeutic use against various malignancies. NK cells discriminate between normal cells and transformed cells via a balance of inhibitory and activating signals induced by interactions between NK cell receptors and target cell ligands. Present study investigates whether expansion of NK cells could augment their anti-myeloma (MM) activity. For NK cell expansion, peripheral blood mononuclear cells from healthy donors and myeloma patients were co-cultured with irradiated K562 cells transfected with 4-1BBL and membrane-bound IL15 (K562-mb15-41BBL). A genome-wide profiling approach was performed to identify gene expression signature in expanded NK (ENK) cells and non-expanded NK cells isolated from healthy donors and myeloma patients. A specific set of genes involved in proliferation, migration, adhesion, cytotoxicity, and activation were up regulated post expansion, also confirmed by flow cytometry. Exp-NK cells killed both allogeneic and autologous primary MM cells more avidly than non-exp-NK cells in vitro. Multiple receptors, particularly NKG2D, natural cytotoxicity receptors, and DNAM-1 contributed to target lysis, via a perforin mediated mechanism. In summary, vigorous expansion and high anti-MM activity both in vitro and in vivo provide the rationale for testing exp-NK cells in a clinical trial for high risk MM. Differential gene expression profile in expanded natural killer (ENK) cells and non-expanded natural killer (NK) cells from healthy donors and myeloma patients Eight healthy donor and eight myeloma patients were used in the study. Non-expanded natural killer (NK) cells were isolated from PBMCs of healthy donors and myeloma patients. Expanded natural killer (ENK) cells were generated from same set of samples as mentioned in expansion protocol. All ENK and NK cells were used for gene expression profiling.

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Expression profile of human natural killer cells co-cultured with P. falciparum infected erythrocytes

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