We show diverse functional studies by flow cytometry of not reported ALPS-like patients, diagnosed in the authors’ laboratories. Immunophenotyping of T, B and NK compartments, protein expression (NKG2D, XIAP, SAP, CTLA4 and LRBA) and phosphorylation assays (AKT, STAT3 and STAT1) could be rapid and useful to direct the diagnosis in some patients. Serial monitoring of serum levels of soluble CD25 (sCD25) (R&D system, Abingdon, UK) could be very useful as a measure of disease activity and response to therapy. For further details, see methods in Supplementary Material Supplementary Table 3
Scd25
Manufactured by R&D Systems
Sourced in United Kingdom, Spain
The SCD25 is a compact digital conductivity and dissolved oxygen meter designed for laboratory use. It features simultaneous measurement of conductivity and dissolved oxygen, with the ability to display parameters such as salinity, total dissolved solids, and oxygen saturation. The device offers user-friendly operation and data storage capabilities.
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Lab products found in correlation
Il 10, by Thermo Fisher Scientific (1 mentions)
Vitamin b12, by Beckman Coulter (1 mentions)
Anti tcrγδ pe, by BD (1 mentions)
Anti hla dr pe, by BD (1 mentions)
Anti cd3 percp5, by BD (1 mentions)
Anti cd19 fitc, by Beckman Coulter (1 mentions)
Anti cd4 apc, by Beckman Coulter (1 mentions)
Anti igd pe, by BD (1 mentions)
Sfasl, by R&D Systems (1 mentions)
Anti cd27 pc5, by Beckman Coulter (1 mentions)
Navios cytometer, by Beckman Coulter (1 mentions)
2 protocols using scd25
1
ALPS-like Patients: Flow Cytometry Profiling
2
Multiparametric Flow Cytometry Analysis
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Proportions of DN T cells were determined in total blood samples by flow cytometry using a Navios Cytometer (Beckman Coulter, Madrid, Spain). Mouse anti-human monoclonal antibodies were used to identify this population: anti-TCRαβ-FITC, anti-TCRγδ-PE, anti-CD3-PerCP5.5 (BD Biosciences, Madrid, Spain), anti-CD8-PeCy7, anti-CD4-APC, and anti-CD45-APC-Alexa Fluor 750 (Beckman Coulter). Activated T-cells and B-cell phenotype were also tested in the ALPS-FASLG patient. Mouse anti-human monoclonal antibodies were used to identify these populations: anti-HLA-DR-PE, anti-CD3-PerCP5.5, anti-IgD-PE (BD Biosciences), anti-CD19-FITC, and anti-CD27-PC5 (Beckman Coulter).
Plasma levels of IL-10 (Bender MedSystems, LabClinics, Madrid, Spain), sCD25 and sFasL (R&D Systems, Vitro, Madrid, Spain) and vitamin B12 (Beckman Coulter) were measured in duplicate by enzyme-linked immunosorbent assay. Serum immunoglobulin levels (IgG, IgA, IgM) were measured by nephelometry (Beckman Coulter).
Plasma levels of IL-10 (Bender MedSystems, LabClinics, Madrid, Spain), sCD25 and sFasL (R&D Systems, Vitro, Madrid, Spain) and vitamin B12 (Beckman Coulter) were measured in duplicate by enzyme-linked immunosorbent assay. Serum immunoglobulin levels (IgG, IgA, IgM) were measured by nephelometry (Beckman Coulter).
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