Experiment Overview
| Repository ID: | FR-FCM-ZYED | Experiment name: | Single cell based phosphorylation profiling identifies alterations in Toll-like receptor 7 and 9 signaling in patients with primary Sjögren?s syndrome | MIFlowCyt score: | 85.90% |
| Primary researcher: | Richard Davies | PI/manager: | Richard Davies | Uploaded by: | Richard Davies |
| Experiment dates: | 2015-09-03 - 2015-10-17 | Dataset uploaded: | Nov 2017 | Last updated: | Apr 2024 |
| Keywords: | [Toll-like receptors] [Sjögren’s syndrome] [extraglandular manifestations] [autoantibodies] [phosphoflow] [type I interferon] | Manuscripts: | |||
| Organizations: |
University of Bergen, Clinical Institute 2, Bergen, (Norway)
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| Purpose: | To gain insights into potential dysfunctional intracellular signalling profiles of pSS patients we conducted an exploratory analysis of MAPK/ERK and JAK/STAT signalling networks in peripheral blood mononuclear cells (PBMC) from 25 female pSS patients and 25 female age-matched healthy donors using phospho-specific flow cytometry. We analyzed unstimulated samples, and samples during a 4 hour time period following activation of Toll-like receptor (TLR) 7 and 9. | ||||
| Conclusion: | We here report increased signaling potentials in PBMCs from pSS patients to TLR7 and -9 stimulation through STAT3 S727 and NF-κB in B cells that correlates with a type I IFN signature. Induction of these pathways could contribute to the generation of a type I IFN signature in pSS. Patients displaying elevated potentiation of aforementioned pathways could therefore benefit from therapies targeting these pathways. | ||||
| Comments: | Stimulation with TLR7 and 9 ligands was conducted using a 4 hour reverse time course. Barcoded samples included a PBMC sample from a single donor (IC) to assess assay variation. Key 9x barcoding stimulus are as follows- POlow + PB low= Unstimulated, POlow + PBmid= IC (unstimulated), POlow + PBhigh= 3 hours, POmid + PBlow= 15 minutes, POmid + PBmid= 60 minutes, POmid + PBhigh = 4 hours, POhigh + PBlow = 30 minutes, POhigh + PBmid = 2 hours, and POhigh + PBhigh= IC (stimulated). | ||||
| Funding: | The authors’ research is supported by the EU H2020 contract HarmonicSS (H2020-SC1-2016-RTD/731944), the Broegelmann Foundation, the Western Norway Regional Health Authorities (grant nr. 912065) and the University of Bergen. | ||||
| Quality control: | An optimized multiplex flow cytometry protocol for the analysis of intracellular signaling in peripheral blood mononuclear cells | The cytometer was routinely calibrated with BD cytometer setup and tracking beads (BD Biosciences). Stimulated and unstimulated PBMC from a single donor was included in each FC barcoded assay to monitor assay variation. | |||
Experiment variables
| Sample Type | |
|---|---|
| · pSS | DC744_Barcode_004.fcs · DC751_Barcode_018.fcs · DC752_Barcode_010.fcs · DC756_Barcode_010.fcs · DC758_Barcode_010.fcs · DC759_Barcode_005.fcs · DC763_Barcode_018.fcs · DC765_Barcode_018.fcs · DC769_Barcode_014.fcs · DC773_Barcode_014.fcs · DC775_Barcode_005.fcs · DC800_Barcode_010.fcs · DC805_Barcode_005.fcs · DC811_Barcode_018.fcs · DC817_Barcode_005.fcs · DC818_Barcode_016.fcs · DC820_Barcode_014.fcs · DC822_Barcode_014.fcs · DC828_Barcode_005.fcs · DC829_Barcode_018.fcs · DC833_Barcode_010.fcs · DC840_Barcode_005.fcs · DC841_Barcode_010.fcs · DC846_Barcode_018.fcs · DC855_Barcode_005.fcs · DC744_Panel 1_001.fcs · DC751_Panel 1_015.fcs · DC752_Panel 1_007.fcs · DC756_Panel 1_007.fcs · DC758_Panel 1_007.fcs · DC759_Panel 1_002.fcs · DC763_Panel 1_015.fcs · DC765_Panel 1_015.fcs · DC769_Panel 1_011.fcs · DC773_Panel 1_011.fcs · DC775_Panel 1_002.fcs · DC800_Panel 1_007.fcs · DC805_Panel 1_002.fcs · DC811_Panel 1_015.fcs · DC817_Panel 1_002.fcs · DC818_Panel 1_013.fcs · DC820_Panel 1_011.fcs · DC822_Panel 1_011.fcs · DC828_Panel 1_002.fcs · DC829_Panel 1_015.fcs · DC833_Panel 1_007.fcs · DC840_Panel 1_002.fcs · DC841_Panel 1_007.fcs · DC846_Panel 1_015.fcs · DC855_Panel 1_002.fcs · DC744_Panel 2_002.fcs · DC751_Panel 2_016.fcs · DC752_Panel 2_008.fcs · DC756_Panel 2_008.fcs · DC758_Panel 2_008.fcs · DC759_Panel 2_003.fcs · DC763_Panel 2_016.fcs · DC765_Panel 2_016.fcs · DC769_Panel 2_012.fcs · DC773_Panel 2_012.fcs · DC775_Panel 2_003.fcs · DC800_Panel 2_008.fcs · DC805_Panel 2_003.fcs · DC811_Panel 2_016.fcs · DC817_Panel 2_003.fcs · DC818_Panel 2_014.fcs · DC820_Panel 2_012.fcs · DC822_Panel 2_012.fcs · DC828_Panel 2_003.fcs · DC829_Panel 2_016.fcs · DC833_Panel 2_008.fcs · DC840_Panel 2_003.fcs · DC841_Panel 2_008.fcs · DC846_Panel 2_016.fcs · DC855_Panel 2_003.fcs · DC744_Panel 3_003.fcs · DC751_Panel 3_017.fcs · DC752_Panel 3_009.fcs · DC756_Panel 3_009.fcs · DC758_Panel 3_009.fcs · DC759_Panel 3_004.fcs · DC763_Panel 3_017.fcs · DC765_Panel 3_017.fcs · DC769_Panel 3_013.fcs · DC773_Panel 3_013.fcs · DC775_Panel 3_004.fcs · DC800_Panel 3_009.fcs · DC805_Panel 3_004.fcs · DC811_Panel 3_017.fcs · DC817_Panel 3_004.fcs · DC818_Panel 3_015.fcs · DC820_Panel 3_013.fcs · DC822_Panel 3_013.fcs · DC828_Panel 3_004.fcs · DC829_Panel 3_017.fcs · DC833_Panel 3_009.fcs · DC840_Panel 3_004.fcs · DC841_Panel 3_009.fcs · DC846_Panel 3_017.fcs · DC855_Panel 3_004.fcs |
| · Healthy donor | C001_Barcode_010.fcs · C11_Barcode_014.fcs · C12_Barcode_014.fcs · C16_Barcode_014.fcs · C19_Barcode_014.fcs · C22(2)_Barcode_005.fcs · C23_Barcode_010.fcs · C25_Barcode_018.fcs · C26_Barcode_014.fcs · C28_Barcode_010.fcs · C31_Barcode_014.fcs · C32_Barcode_010.fcs · C39_Barcode_005.fcs · C40_Barcode_020.fcs · C43_Barcode_005.fcs · C45_Barcode_018.fcs · C46_Barcode_018.fcs · C5_Barcode_005.fcs · C6_Barcode_010.fcs · M012_Barcode_010.fcs · M014_Barcode_005.fcs · M015_Barcode_014.fcs · M019_Barcode_018.fcs · M023_Barcode_010.fcs · M024_Barcode_018.fcs · C001_Panel 1_007.fcs · C11_Panel 1_011.fcs · C12_Panel 1_011.fcs · C16_Panel 1_011.fcs · C19_Panel 1_011.fcs · C22(2)_Panel 1_002.fcs · C23_Panel 1_007.fcs · C25_Panel 1_015.fcs · C26_Panel 1_011.fcs · C28_Panel 1_007.fcs · C31_Panel 1_011.fcs · C32_Panel 1_001.fcs · C39_Panel 1_002.fcs · C40_Panel 1_017.fcs · C43_Panel 1_002.fcs · C45_Panel 1_015.fcs · C46_Panel 1_015.fcs · C5_Panel 1_002.fcs · C6_Panel 1_007.fcs · M012_Panel 1_007.fcs · M014_Panel 1_002.fcs · M015_Panel 1_011.fcs · M019_Panel 1_015.fcs · M023_Panel 1_007.fcs · M024_Panel 1_015.fcs · C001_Panel 2_008.fcs · C11_Panel 2_012.fcs · C12_Panel 2_012.fcs · C16_Panel 2_012.fcs · C19_Panel 2_012.fcs · C22(2)_Panel 2_003.fcs · C23_Panel 2_008.fcs · C25_Panel 2_016.fcs · C26_Panel 2_012.fcs · C28_Panel 2_008.fcs · C31_Panel 2_012.fcs · C32_Panel 2_008.fcs · C39_Panel 2_003.fcs · C40_Panel 2_018.fcs · C43_Panel 2_003.fcs · C45_Panel 2_016.fcs · C46_Panel 2_016.fcs · C5_Panel 2_003.fcs · C6_Panel 2_008.fcs · M012_Panel 2_008.fcs · M014_Panel 2_003.fcs · M015_Panel 2_012.fcs · M019_Panel 2_016.fcs · M023_Panel 2_008.fcs · M024_Panel 2_016.fcs · C001_Panel 3_009.fcs · C11_Panel 3_013.fcs · C12_Panel 3_013.fcs · C16_Panel 3_013.fcs · C19_Panel 3_013.fcs · C22(2)_Panel 3_004.fcs · C23_Panel 3_009.fcs · C25_Panel 3_017.fcs · C26_Panel 3_013.fcs · C28_Panel 3_009.fcs · C31_Panel 3_013.fcs · C32_Panel 3_009.fcs · C39_Panel 3_004.fcs · C40_Panel 3_019.fcs · C43_Panel 3_004.fcs · C45_Panel 3_017.fcs · C46_Panel 3_017.fcs · C5_Panel 3_004.fcs · C6_Panel 3_009.fcs · M012_Panel 3_009.fcs · M014_Panel 3_004.fcs · M015_Panel 3_013.fcs · M019_Panel 3_017.fcs · M023_Panel 3_009.fcs · M024_Panel 3_017.fcs |
| Individuals | |
|---|---|
| · C001 | C001_Barcode_010.fcs · C001_Panel 1_007.fcs · C001_Panel 2_008.fcs · C001_Panel 3_009.fcs |
| · C11 | C11_Barcode_014.fcs · C11_Panel 1_011.fcs · C11_Panel 2_012.fcs · C11_Panel 3_013.fcs |
| · C12 | C12_Barcode_014.fcs · C12_Panel 1_011.fcs · C12_Panel 2_012.fcs · C12_Panel 3_013.fcs |
| · C16 | C16_Barcode_014.fcs · C16_Panel 1_011.fcs · C16_Panel 2_012.fcs · C16_Panel 3_013.fcs |
| · C19 | C19_Barcode_014.fcs · C19_Panel 1_011.fcs · C19_Panel 2_012.fcs · C19_Panel 3_013.fcs |
| · C22(2) | C22(2)_Barcode_005.fcs · C22(2)_Panel 1_002.fcs · C22(2)_Panel 2_003.fcs · C22(2)_Panel 3_004.fcs |
| · C23 | C23_Barcode_010.fcs · C23_Panel 1_007.fcs · C23_Panel 2_008.fcs · C23_Panel 3_009.fcs |
| · C25 | C25_Barcode_018.fcs · C25_Panel 1_015.fcs · C25_Panel 2_016.fcs · C25_Panel 3_017.fcs |
| · C26 | C26_Barcode_014.fcs · C26_Panel 1_011.fcs · C26_Panel 2_012.fcs · C26_Panel 3_013.fcs |
| · C28 | C28_Barcode_010.fcs · C28_Panel 1_007.fcs · C28_Panel 2_008.fcs · C28_Panel 3_009.fcs |
| · C31 | C31_Barcode_014.fcs · C31_Panel 1_011.fcs · C31_Panel 2_012.fcs · C31_Panel 3_013.fcs |
| · C32 | C32_Barcode_010.fcs · C32_Panel 1_001.fcs · C32_Panel 2_008.fcs · C32_Panel 3_009.fcs |
| · C39 | C39_Barcode_005.fcs · C39_Panel 1_002.fcs · C39_Panel 2_003.fcs · C39_Panel 3_004.fcs |
| · C40 | C40_Barcode_020.fcs · C40_Panel 1_017.fcs · C40_Panel 2_018.fcs · C40_Panel 3_019.fcs |
| · C43 | C43_Barcode_005.fcs · C43_Panel 1_002.fcs · C43_Panel 2_003.fcs · C43_Panel 3_004.fcs |
| · C45 | C45_Barcode_018.fcs · C45_Panel 1_015.fcs · C45_Panel 2_016.fcs · C45_Panel 3_017.fcs |
| · C46 | C46_Barcode_018.fcs · C46_Panel 1_015.fcs · C46_Panel 2_016.fcs · C46_Panel 3_017.fcs |
| · C5 | C5_Barcode_005.fcs · C5_Panel 1_002.fcs · C5_Panel 2_003.fcs · C5_Panel 3_004.fcs |
| · C6 | C6_Barcode_010.fcs · C6_Panel 1_007.fcs · C6_Panel 2_008.fcs · C6_Panel 3_009.fcs |
| · DC744 | DC744_Barcode_004.fcs · DC744_Panel 1_001.fcs · DC744_Panel 2_002.fcs · DC744_Panel 3_003.fcs |
| · DC751 | DC751_Barcode_018.fcs · DC751_Panel 1_015.fcs · DC751_Panel 2_016.fcs · DC751_Panel 3_017.fcs |
| · DC752 | DC752_Barcode_010.fcs · DC752_Panel 1_007.fcs · DC752_Panel 2_008.fcs · DC752_Panel 3_009.fcs |
| · DC756 | DC756_Barcode_010.fcs · DC756_Panel 1_007.fcs · DC756_Panel 2_008.fcs · DC756_Panel 3_009.fcs |
| · DC758 | DC758_Barcode_010.fcs · DC758_Panel 1_007.fcs · DC758_Panel 2_008.fcs · DC758_Panel 3_009.fcs |
| · DC759 | DC759_Barcode_005.fcs · DC759_Panel 1_002.fcs · DC759_Panel 2_003.fcs · DC759_Panel 3_004.fcs |
| · DC763 | DC763_Barcode_018.fcs · DC763_Panel 1_015.fcs · DC763_Panel 2_016.fcs · DC763_Panel 3_017.fcs |
| · DC765 | DC765_Barcode_018.fcs · DC765_Panel 1_015.fcs · DC765_Panel 2_016.fcs · DC765_Panel 3_017.fcs |
| · DC769 | DC769_Barcode_014.fcs · DC769_Panel 1_011.fcs · DC769_Panel 2_012.fcs · DC769_Panel 3_013.fcs |
| · DC773 | DC773_Barcode_014.fcs · DC773_Panel 1_011.fcs · DC773_Panel 2_012.fcs · DC773_Panel 3_013.fcs |
| · DC775 | DC775_Barcode_005.fcs · DC775_Panel 1_002.fcs · DC775_Panel 2_003.fcs · DC775_Panel 3_004.fcs |
| · DC800 | DC800_Barcode_010.fcs · DC800_Panel 1_007.fcs · DC800_Panel 2_008.fcs · DC800_Panel 3_009.fcs |
| · DC805 | DC805_Barcode_005.fcs · DC805_Panel 1_002.fcs · DC805_Panel 2_003.fcs · DC805_Panel 3_004.fcs |
| · DC811 | DC811_Barcode_018.fcs · DC811_Panel 1_015.fcs · DC811_Panel 2_016.fcs · DC811_Panel 3_017.fcs |
| · DC817 | DC817_Barcode_005.fcs · DC817_Panel 1_002.fcs · DC817_Panel 2_003.fcs · DC817_Panel 3_004.fcs |
| · DC818 | DC818_Barcode_016.fcs · DC818_Panel 1_013.fcs · DC818_Panel 2_014.fcs · DC818_Panel 3_015.fcs |
| · DC820 | DC820_Barcode_014.fcs · DC820_Panel 1_011.fcs · DC820_Panel 2_012.fcs · DC820_Panel 3_013.fcs |
| · DC822 | DC822_Barcode_014.fcs · DC822_Panel 1_011.fcs · DC822_Panel 2_012.fcs · DC822_Panel 3_013.fcs |
| · DC828 | DC828_Barcode_005.fcs · DC828_Panel 1_002.fcs · DC828_Panel 2_003.fcs · DC828_Panel 3_004.fcs |
| · DC829 | DC829_Barcode_018.fcs · DC829_Panel 1_015.fcs · DC829_Panel 2_016.fcs · DC829_Panel 3_017.fcs |
| · DC833 | DC833_Barcode_010.fcs · DC833_Panel 1_007.fcs · DC833_Panel 2_008.fcs · DC833_Panel 3_009.fcs |
| · DC840 | DC840_Barcode_005.fcs · DC840_Panel 1_002.fcs · DC840_Panel 2_003.fcs · DC840_Panel 3_004.fcs |
| · DC841 | DC841_Barcode_010.fcs · DC841_Panel 1_007.fcs · DC841_Panel 2_008.fcs · DC841_Panel 3_009.fcs |
| · DC846 | DC846_Barcode_018.fcs · DC846_Panel 1_015.fcs · DC846_Panel 2_016.fcs · DC846_Panel 3_017.fcs |
| · DC855 | DC855_Barcode_005.fcs · DC855_Panel 1_002.fcs · DC855_Panel 2_003.fcs · DC855_Panel 3_004.fcs |
| · M012 | M012_Barcode_010.fcs · M012_Panel 1_007.fcs · M012_Panel 2_008.fcs · M012_Panel 3_009.fcs |
| · M014 | M014_Barcode_005.fcs · M014_Panel 1_002.fcs · M014_Panel 2_003.fcs · M014_Panel 3_004.fcs |
| · M015 | M015_Barcode_014.fcs · M015_Panel 1_011.fcs · M015_Panel 2_012.fcs · M015_Panel 3_013.fcs |
| · M019 | M019_Barcode_018.fcs · M019_Panel 1_015.fcs · M019_Panel 2_016.fcs · M019_Panel 3_017.fcs |
| · M023 | M023_Barcode_010.fcs · M023_Panel 1_007.fcs · M023_Panel 2_008.fcs · M023_Panel 3_009.fcs |
| · M024 | M024_Barcode_018.fcs · M024_Panel 1_015.fcs · M024_Panel 2_016.fcs · M024_Panel 3_017.fcs |
