Experiment Overview

Repository ID: FR-FCM-ZYR5 Experiment name: SOM3B Analysis on PMA/Ionomycin stimulated human peripheral whole blood MIFlowCyt score: 61.74%
Primary researcher: Sam Kimmey PI/manager: Sam Kimmey Uploaded by: Sam Kimmey
Experiment dates: 2018-10-09 - Dataset uploaded: Oct 2018 Last updated: May 2019
Keywords: [Activation markers] [whole blood] [whole blood intracellular staining] [CyTOF] [PMA] [ionomycin] [translation activity] [transcription activity] Manuscripts: [30862852] [PMC6414513]
Organizations: Stanford University, Stanford, CA (USA)
Purpose: To assess global activity of de novo transcription and translation upon phorbol 12-myristate 13-acetate (PMA)/ionomycin stimulation. Whole blood from one donor was collected and split into a time-course series to stimulate cells between 15 minutes and 5 hours. A mass-cytometry panel was designed to isolate peripheral mononuclear cells into major phenotype groups across the stimulation time course. Additional functional parameters that were measured in parallel indicate the activation state and characteristics of cells across stimulation time.
Conclusion: Peripheral mononuclear cells display a dynamic response in transcription and translation activity upon stimulation with PMA/ionomycin. A strong activation of de novo transcription activity is an immediate response in the B-cell sub-population, with is followed by transient activation of de novo protein synthesis, along with canonical markers of activation, such as the surface epitope CD69.
Comments: None
Funding: Not disclosed
Quality control: Samples of whole blood from the same donor were not labeled with SOM3B reagents, or treated with specific inhibitors of transcription or translation to confirm specificity of SOM3B labeling.

Experiment variables

· Baseline No_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · No_Stim_MNCs_CD19_20+ B Cells.fcs · No_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · No_Stim_MNCs_CD4 T Cells.fcs · No_Stim_MNCs_CD4neg T Cells.fcs · No_Stim_MNCs_CD56+ NK.fcs · No_Stim_MNCs_Classical Monocytes_CD14+.fcs · No_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · No_Stim_MNCs_cDCs.fcs
· 15 min 0_25h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 0_25h_Stim_MNCs_CD19_20+ B Cells.fcs · 0_25h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 0_25h_Stim_MNCs_CD4 T Cells.fcs · 0_25h_Stim_MNCs_CD4neg T Cells.fcs · 0_25h_Stim_MNCs_CD56+ NK.fcs · 0_25h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 0_25h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 0_25h_Stim_MNCs_cDCs.fcs
· 30 min 0_5h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 0_5h_Stim_MNCs_CD19_20+ B Cells.fcs · 0_5h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 0_5h_Stim_MNCs_CD4 T Cells.fcs · 0_5h_Stim_MNCs_CD4neg T Cells.fcs · 0_5h_Stim_MNCs_CD56+ NK.fcs · 0_5h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 0_5h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 0_5h_Stim_MNCs_cDCs.fcs
· 1h 1h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 1h_Stim_MNCs_CD19_20+ B Cells.fcs · 1h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 1h_Stim_MNCs_CD4 T Cells.fcs · 1h_Stim_MNCs_CD4neg T Cells.fcs · 1h_Stim_MNCs_CD56+ NK.fcs · 1h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 1h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 1h_Stim_MNCs_cDCs.fcs
· 2h 2h_Stim_CD4neg T Cells.fcs · 2h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 2h_Stim_MNCs_CD19_20+ B Cells.fcs · 2h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 2h_Stim_MNCs_CD4 T Cells.fcs · 2h_Stim_MNCs_CD56+ NK.fcs · 2h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 2h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 2h_Stim_MNCs_cDCs.fcs
· 5h 5h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 5h_Stim_MNCs_CD19_20+ B Cellsfcs.fcs · 5h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 5h_Stim_MNCs_CD4 T Cells.fcs · 5h_Stim_MNCs_CD4neg T Cells.fcs · 5h_Stim_MNCs_CD56+ NK.fcs · 5h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 5h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 5h_Stim_MNCs_cDCs.fcs

· pDCs 0_25h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 0_5h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 1h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 2h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · 5h_Stim_MNCs_CD123+ HLADR+ pDCs.fcs · No_Stim_MNCs_CD123+ HLADR+ pDCs.fcs
· B cells 0_25h_Stim_MNCs_CD19_20+ B Cells.fcs · 0_5h_Stim_MNCs_CD19_20+ B Cells.fcs · 1h_Stim_MNCs_CD19_20+ B Cells.fcs · 2h_Stim_MNCs_CD19_20+ B Cells.fcs · 5h_Stim_MNCs_CD19_20+ B Cellsfcs.fcs · No_Stim_MNCs_CD19_20+ B Cells.fcs
· Basophils 0_25h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 0_5h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 1h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 2h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · 5h_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs · No_Stim_MNCs_CD38+ CD123+ HLADCneg Basophils.fcs
· CD4 T Cells 0_25h_Stim_MNCs_CD4 T Cells.fcs · 0_5h_Stim_MNCs_CD4 T Cells.fcs · 1h_Stim_MNCs_CD4 T Cells.fcs · 2h_Stim_MNCs_CD4 T Cells.fcs · 5h_Stim_MNCs_CD4 T Cells.fcs · No_Stim_MNCs_CD4 T Cells.fcs
· CD4 negative (CD8) T cells 0_25h_Stim_MNCs_CD4neg T Cells.fcs · 0_5h_Stim_MNCs_CD4neg T Cells.fcs · 1h_Stim_MNCs_CD4neg T Cells.fcs · 2h_Stim_CD4neg T Cells.fcs · 5h_Stim_MNCs_CD4neg T Cells.fcs · No_Stim_MNCs_CD4neg T Cells.fcs
· NK 0_25h_Stim_MNCs_CD56+ NK.fcs · 0_5h_Stim_MNCs_CD56+ NK.fcs · 1h_Stim_MNCs_CD56+ NK.fcs · 2h_Stim_MNCs_CD56+ NK.fcs · 5h_Stim_MNCs_CD56+ NK.fcs · No_Stim_MNCs_CD56+ NK.fcs
· cDCs 0_25h_Stim_MNCs_cDCs.fcs · 0_5h_Stim_MNCs_cDCs.fcs · 1h_Stim_MNCs_cDCs.fcs · 2h_Stim_MNCs_cDCs.fcs · 5h_Stim_MNCs_cDCs.fcs · No_Stim_MNCs_cDCs.fcs
· CD14+ Monocyte 0_25h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 0_5h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 1h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 2h_Stim_MNCs_Classical Monocytes_CD14+.fcs · 5h_Stim_MNCs_Classical Monocytes_CD14+.fcs · No_Stim_MNCs_Classical Monocytes_CD14+.fcs
· CD16+ Monocyte 0_25h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 0_5h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 1h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 2h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · 5h_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs · No_Stim_MNCs_Non-Classical Monocytes_CD16+.fcs

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