Unknown,Transcriptomics,Genomics,Proteomics

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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. 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.

ORGANISM(S): Homo sapiens

SUBMITTER: Ivan Imaz-Rosshandler

PROVIDER: E-GEOD-69252 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Ex vivo activation and expansion of natural killer (NK) cells is a strategy to produce sufficient numbers of these effector cells for adoptive immunotherapy. Therefore we aimed at the development of an automated, clinical scale NK cell expansion process and compared NK cells after manual and automated expansion.We found only a small subset of 124 genes that significantly varied between both sample groups. These genes were associated with movement of leukocytes were identified including a group of genes for NK cell movement (CMKLR1, CX3CR1, S1PR5, GNLY and CXCR1) which were slightly higher expressed after automated compared to manual expansion. Nevertheless, the expansion profiles of NK cells after automated or manual expansion were rather similar in comparison to the remarkable difference between primary and expanded NK cells in general represented by the vast majority of regulated genes between the analyzed sample groups. Pathway analysis revealed that most of regulated genes upon expansion were associated with cell cycle regulation, DNA replication, DNA recombination and DNA repair, regulation of apoptosis and cell survival as well as cytokine signaling. Furthermore, the expression of many NK cell relevant markers was changed upon expansion with the strongest effects on up regulation of TRAIL and FASL, inhibitory TIGIT and chemokine receptors CCR2, CCR5 and CXCR6. In addition, granzyme M was down regulated but other important effector molecules like TNFa, perforin and granzymes A, B and K were up regulated. In summary we could show that manual and automatically expanded NK cells show a similar expansion profile while the differ significantly from primary NK cells. Up regulation of many NK cell relevant markers indicate for an enhanced NK cell functionaility after ex vivo activation and expansion. 24 samples were analyzed in total. 6 biological replicates represented by cells from different donors. 4 different samples per donor: freshly isolated NK cells (d0) and NK cells expanded for 14 days under 3 different conditions: in co-culture with EBV-LCL feeder cells and 500 U/mL IL2 either cultivated manually in T75 flasks (T) or automatically using the CliniMACS Prodigy system (P); NK cells cultured with 500 U/ml IL2 without feeder cells (I)

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

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