Antibodies used in this study are as follows: CD3 (clone SP-34–2; BD Biosciences), CXCR5 (MU5UBEE; eBioscience), GagCM9 tetramer (NIH tetramer core), CD28 (CD28.2; eBioscience), CD95 (DX2; BD Biosciences), CD279 (PD-1; clone EH12.2H7; BioLegend), CD8 (SK1; BD Bioscience), Ki67 (B56; BD Biosciences), CD4 (L200; BD Biosciences), GrzB (GB11; BD Biosciences), perforin (Pf-344; Mabtech), FoxP3 (206D; BioLegend), HLA-DR (L243; BioLegend), IFNλ (B27; BD Biosciences), TNFα (MAb11; BD Biosciences), and IL-2 (MQ1- 17H12; BD Biosciences).
Il 2 mq1 17h12
Manufactured by BD
Sourced in United States
IL-2 (MQ1-17H12) is a monoclonal antibody that binds to and detects human interleukin-2 (IL-2). IL-2 is a cytokine that plays a crucial role in the activation and proliferation of T cells, which are important for immune responses.
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Lab products found in correlation
Ionomycin, by Merck Group (3 mentions)
Phorbol myristate acetate (pma), by Merck Group (2 mentions)
Cd3 clone sp34 2, by BD (1 mentions)
Cd8 sk1, by BD (1 mentions)
Cd279 pd1 clone eh12.2h7, by BioLegend (1 mentions)
Cxcr5 mu5ubee, by Thermo Fisher Scientific (1 mentions)
Cd28 cd28.2, by Thermo Fisher Scientific (1 mentions)
Cd95 dx2, by BD (1 mentions)
Ki67 b56, by BD (1 mentions)
Cd4 l200, by BD (1 mentions)
Perforin pf 344, by Mabtech (1 mentions)
Hla dr l243, by BioLegend (1 mentions)
Tnfα mab11, by BD (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Monensin protein transport inhibitor, by BD (1 mentions)
Golgistop, by BD (1 mentions)
Facscalibur, by Tree Star (1 mentions)
Interleukin 2 (il 2), by Cell Signaling Technology (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Kaluza software, by Beckman Coulter (1 mentions)
4 protocols using il 2 mq1 17h12
1
Multi-Marker Immune Profiling Protocol
2
Antigen-specific CD8+ T cell Assay
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After 7-days of expansion with VIPR particles, the T cell cultures were stimulated for 6 h with 20 μg/ml peptide antigen (> 95% purity) specific for the VIPR particle expanded CD8+ population (CMV pp65: NLVPMVATV; SARS-CoV-2 S protein 269–277 YLQPRTFLL), all peptides were synthesized and obtained from ProImmune, Ltd. After 1-h of peptide stimulation, GolgiStop solution was added (containing Monensin protein transport inhibitor) to block intracellular protein transport (BD Bioscience). As a positive control for cytokine production, T cells were also stimulated for 6 h with 50 ng/ml PMA and 1 μg/ml Ionomycin (Sigma). T cells were then harvested, and stained for surface markers CD8 (HIT8A, 1:80), CD107a (H4A3 1:50), then fixed and permeabilized using BD Cytofix/Cytoperm Fixation/Permeabilization Solution (ThermoFisher). Cells were then stained for surface markers followed by intracellular cytokines using antibodies specific for IFN-y (4S.B3, 1:40) ), GzmB (QA16A02 1:50), Perforim A (dG9 1:50) (Biolegend) IL-2 (MQ1-17H12, 1:40) (BD Bioscience), or TNF-a (MAb11, 1:40) (ThermoFisher). Flow analysis was carried out on a Fortessa flow cytometer (BD Bioscience) and data analyzed and flow cytometry figures generated using FlowJo 10 software (BD Biosciences).
3
Cytokine profiling of stimulated PBMCs
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PBMCs were stimulated with 5 ng/ml IL-2 (Cell Signaling), 50 ng/ml PMA (Merck), 1 μg/ml ionomycin (Sigma Aldrich), and GolgiStop (BD Biosciences) was added for the final 5 hours. PBMCs were stained with anti-human TCRγδ and CXCR5. PBMCs were then fixed using a BD Perm/Fix intracellular staining kit. PBMCs were then stained with IL-4 (MP4-25D2), IL-10 (JES3-9D7), IFNγ (4S.B3) (all from Biolegend, San Diego, CA, USA) and IL-2 (MQ1-17H12, BD Biosciences, San Diego, CA, USA) at room temperature for 30 min at dark. Data were collected by flow cytometry on a FACScalibur and were analyzed with FlowJo software (TreeStar).
4
Comprehensive Immune Cell Profiling
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For cell surface staining, the following fluorochrome-conjugated monoclonal antibodies were used CD3(UCHT1), CD4(13B8.2), CD8(B9.11), CD19(J3-119), CD24(ALB9), CD38(LS198-4-3), CD45(J.33), CD45RO(UCHL1), CD127(R34.34) and IgD(IADB6) (all from Beckman-Coulter, Mijdrecht, The Netherlands), CD25(M-A251), CCR4(1G1), CCR6(11A9) and CXCR3(1C6/CXR3) (BD Biosciences, Erembodegem, Belgium), and BAFF-R(11C1) (BioLegend, Uithoorn, The Netherlands). Intracellular analysis of IL-2(MQ1-17H12) (BD Biosciences), IL-4(8D4–8), IL-17(EBIO64CAP17), IFNγ(4S.B3) (eBioscience, San Diego, CA, USA), and TNFα(Mab11) (Dako, Glostrup, Denmark) was performed after fixation and permeabilization, using Fix and Perm reagent (eBioscience). Before intracellular cytokine measurement, the cells were stimulated for 4 hours with PMA (12.5 ng/ml), ionomycin (500 ng/ml) and Brefeldin A (5 µg/ml; Sigma-Aldrich, Zwijndrecht, The Netherlands).
The cell phenotype was analyzed by five-color (FC500) or ten-color flow cytometry (Navios), and data were analyzed using Kaluza software (all from Beckman-Coulter). Isotype controls or unstained cells were used for gate settings. Cell populations >0.1% of the CD45+ lymphocyte population with a threshold of more than 50 cells were considered reliable.
The cell phenotype was analyzed by five-color (FC500) or ten-color flow cytometry (Navios), and data were analyzed using Kaluza software (all from Beckman-Coulter). Isotype controls or unstained cells were used for gate settings. Cell populations >0.1% of the CD45+ lymphocyte population with a threshold of more than 50 cells were considered reliable.
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