The TILs from lung cancer patients and HD-PBMCs from healthy donors were incubated in 24-well plates precoated with 1 µg/mL α-CD3 (cat. 317315; Biolegend) in complete X-VIVO 15 medium contained 1 µg/mL α-CD28 (cat. 302923; Biolegend) and 1000 UI/mL IL-2, with or without 7 µM TWS119 (cat. S1590; Selleckchem) for 9 days. TILs and HD-PBMCs cells were harvested on days 3, 5, 7, 9, and then stained for flow cytometry. The following panel of mouse anti-human mAbs, all purchased from BD Biosciences (San Jose, CA) or eBioscience (San Diego, CA), were used: anti-human CD3-APC.Cy7 (BD, 557832, SK7), anti-human CD4-Percp.Cy5.5 (BD, 560650, RPA-T4), anti-human CD45RA-FITC (11-0458-42, HI100; eBioscience), anti-human CCR7-AF700 (BD, 561143, 150503), anti-human CD62L-PE-CF594 (BD, 562301, Dreg-56), anti-human CD95-PE.Cy7 (BD, 561633, DX2), and anti-human CD122-PE (BD, 554525). The cell data were acquired using a BD LSRFortessa analytical flow cytometer. Unstained and single fluorochrome-stained cells were served as controls to provide accurate compensation and data analysis. Cells were recorded per sample and the data were analyzed with FlowJo (version 10).
Anti human cd4 percp
Manufactured by BD
Sourced in United States
The Anti-human CD4-Percp is a lab equipment product that is used to detect and measure the presence of CD4 cells, a type of T-lymphocyte, in a biological sample. It utilizes the Peridinin-Chlorophyll-Protein (PerCP) fluorescent dye to label the CD4 cells, allowing for their identification and quantification through flow cytometry or similar analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
αcd28, by BioLegend (1 mentions)
Anti human cd3 apc cy7, by BD (1 mentions)
Anti human cd95 pe cy7, by BD (1 mentions)
Anti human cd122 pe, by BD (1 mentions)
Lsrfortessa analytical flow cytometer, by BD (1 mentions)
Anti human cd8 fitc, by BD (1 mentions)
Anti human cd3 apc h7, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Anti cd69 pe, by BD (1 mentions)
3 protocols using anti human cd4 percp
1
Characterization of T-cell Subset Dynamics
2
Phenotyping T-cell Activation in Vaccination
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Fresh heparinized peripheral whole blood (150 µL) was stained with anti-human CD3-APC-H7, anti-human CD4-PerCP, anti-human CD8-FITC and anti-CD69-PE (BD Pharmingen, USA) at room temperature for 15 min in the dark. Red cells were lysed with 1x BD FACS lysis buffer (BD Biosciences, USA) for 10 min in the dark. After centrifugation at 450 × g for 10 min, samples were washed once with phosphate-buffered-saline (PBS) before analysis using a BD FACSCantoTM II flow cytometer (BD Bioscience, USA). Activated (CD69+) T cells were determined as shown in Supplementary Figure S1 . To account for the possibility of asymptomatic infection at CV0 (and hence elevated Th responder cell numbers), we measured the changes in CD4 T cell numbers from CV0-CV1 or from CV1-CV2. Participants were sub-grouped into non-T helper (Non-Th) responders (115/151 [76.2%]) and Th responders (36/151 [23.8%]) based on the increased numbers of helper lymphocytes between from CV0 to CV1 and/or CV2. An average of a > 1.47 fold (±SD) change in Th numbers (non-responders vs responders) was taken as a vaccine-dependent difference.
3
CD4+ T Cell Immunophenotyping Protocol
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Peripheral blood mononuclear cells were sampled at the beginning of this study. The cells were washed in PBS and suspended in staining buffer with antihuman CD4-PerCP (BD Biosciences, Bedford, MA, USA) and
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