Affymetrix GeneChip Human Genome HG-U133A [HG-U133A]

Affymetrix GeneChip Human Genome HG-U133B [HG-U133B]

Gene and protein characteristics reflect functional diversity of CD56dim and CD56bright NK cells

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Human NK cells were sorted into CD56dim and CD56bright NK cell subpopulations. In order to define characteristics of both populations gene profiling was performed using Affymetrix arrays U133a and U133B.

Activation

Cell type

Transcription profiling of human NK cells sorted into CD56dim and CD56bright NK cell subpopulations

Human NK cells were sorted into CD56dim and CD56bright NK cell subpopulations. In order to define characteristics of both populations gene profiling was performed using Affymetrix arrays U133a and U133B.


Affymetrix:Protocol:ExpressionStat

Title: Affymetrix CHP Analysis (ExpressionStat). Description:

Affymetrix:Protocol:Hybridization-EukGE-WS2v3

Title: Affymetrix EukGE-WS2v3 Hybridization. Description:

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-9437

P-MEXP-9462

White blood cells were flushed with PBS from lymphocyte filters (Fresenius-Filter) used for preparation of erythrocyte concentrates for transfusion. The recovered white blood cells were then separated by Ficoll-Hypaque centrifugation. PBMC from the interphase were washed twice and then depleted using a cocktail of monoclonal antibodies against CD3 (purified OKT3 hybridoma supernatant), CD14, CD19 (Immunotools, Frisoythe, Germany), and Dynabeads® M450 (Dynal, Hamburg, Germany). Finally, cells were stained with CD56PE and CD3FITC and sorted with FACStar (Becton Dickinson, Heidelberg, Germany) according to expression density of CD56 antigen into CD3-CD56dimNK cells. Purity of the sorted NK cells was reanalyzed by flow cytometry (FACSCalibur, BD Pharmingen, Heidelberg, Germany).

P-MEXP-9463

White blood cells were flushed with PBS from lymphocyte filters (Fresenius-Filter) used for preparation of erythrocyte concentrates for transfusion. The recovered white blood cells were then separated by Ficoll-Hypaque centrifugation. PBMC from the interphase were washed twice and then depleted using a cocktail of monoclonal antibodies against CD3 (purified OKT3 hybridoma supernatant), CD14, CD19 (Immunotools, Frisoythe, Germany), and Dynabeads® M450 (Dynal, Hamburg, Germany). Finally, cells were stained with CD56PE and CD3FITC and sorted with FACStar (Becton Dickinson, Heidelberg, Germany) according to expression density of CD56 antigen into CD3-CD56bright NK cells. Purity of the sorted NK cells was reanalyzed by flow cytometry (FACSCalibur, BD Pharmingen, Heidelberg, Germany).

P-MEXP-9464

P-MEXP-9465

P-MEXP-9512

P-MEXP-9513

RNA of the cell samples were determined. Equal RNA amounts of all samples were mixed.

Transcription profiling of human NK cells sorted into CD56dim and CD56bright NK cell subpopulations

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