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Gene expression profiling in NK cells of patients infected with Leishmania mexicana

ABSTRACT: The aim of this study was to identify differences in the NK-cell response towards Leishmania mexicana lipophosphoglycan (LPG) between patients with localized (LCL) and diffuse (DCL) cutaneous leishmaniasis through gene expression profiling, in an attempt to pinpoint alterations in the signaling pathways responsible for the NK-cell dysfunction in patients with DCL. To determine the gene expression profiling in non stimulated and LPG-stimulated NK cells we include samples of controls, LCL and DCL patients. We performed microarrays (Human Gene 1.0 ST, Affymetrix) to identify differentially expressed transcripts between non stimulated and LPG-stimulated NK cells between controls, LCL and DCL samples.

ORGANISM(S): Homo sapiens

PROVIDER: GSE69252 | GEO | 2016/01/01

SECONDARY ACCESSION(S): PRJNA285024

REPOSITORIES: GEO

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Gene expression profiling in NK cells of patients infected with Leishmania mexicana

Project description:The aim of this study was to identify differences in the NK-cell response towards Leishmania mexicana lipophosphoglycan (LPG) between patients with localized (LCL) and diffuse (DCL) cutaneous leishmaniasis through gene expression profiling, in an attempt to pinpoint alterations in the signaling pathways responsible for the NK-cell dysfunction in patients with DCL. To determine the gene expression profiling in non stimulated and LPG-stimulated NK cells we include samples of controls, LCL and DCL patients. We performed microarrays (Human Gene 1.0 ST, Affymetrix) to identify differentially expressed transcripts between non stimulated and LPG-stimulated NK cells between controls, LCL and DCL samples. Human NK cells from controls, LCL and DCL patients were divided in two conditions: non stimulated and LPG-stimulated (6h) cells, different number of biological replicates were analyzed: four replicates for controls, two replicates for LCL and 3 replicates for DCL patients samples with a total of 18 microarrays.

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Project description:We used Illumina sequencing of poly-A selected RNA of Leishmania mexicana (WHO strain MNYC/BZ/62/M379) culture-adapted promastigotes (PRO), axenic amastigotes (AXA) and intracellular amastigotes (AMA) in mouse bone marrow derived macrophages (BMDM), 24h after infection, to map 5' and 3' ends of Leishmania transcripts and determine transcript abundances. The AMA samples were prepared from total RNA of infected macrophages thus containing a mixture of leishmanial and murine RNA transcripts. We also sequenced poly-A selected RNA from uninfected BMDMs. Three biological replicates per sample.

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