Optimal Method to Stimulate Cytokine Production and Its Use in ...
PMA/ionomycin was the optimal stimulant in our study, and could significantly increase production of five cytokines in 6 h without any significant damage to immune cells. PMA activates protein kinase C, while ionomycin is a calcium ionophore, and stimulation with these compounds bypasses the T cell membrane receptor complex and will lead to ...
Optimization of stimulation and staining conditions for intracellular ...
2021年1月1日 · We investigated ideal stimulation and staining conditions for cytokine producing T cells and monocytes in whole blood. PMA + Ionomycin was the ideal stimulant for cytokine-producing T cells, LPS was the best for cytokine-producing monocytes.
Concomitant evaluation of PMA+ionomycin‐induced kinase …
2014年1月24日 · We examined intracellular ERK1/2 phosphorylation and IFN-γ production by CD4+ and CD8+ T cells upon polyclonal stimulation with PMA and ionomycin, while monitoring expression of the cytolytic molecule perforin and the T cell activation marker CD38.
Stimulation of Cytokine Production in Immune Cells
Stimulation reagents, phorbol 12-myristate 13-acetate (PMA), ionomycin, brefeldin A, and monensin are useful to activate transcription factors for intracellular signaling and production of cytokines of many different immune cell types.
Calcium nanoparticles target and activate T cells to enhance
2024年11月21日 · In this study, we investigate T cell activation using 12-myristate 13-acetate (PMA)-encapsulated CaCO 3 nanoparticles. We find that anti-PD-1 antibody-conjugated CaCO 3 nanoparticles can be...
Comparison of two commonly used methods for stimulating T cells
The current study aimed to determine the optimal conditions for efficient T cell amplification by two commonly used methods involving CD3/CD28 antibody and phytohemagglutinin (PHA), respectively. Results: Orthogonal design and CCK8 assay showed that 5 μg/mL CD3, 5 μg/mL CD28, and 100 ng/mL IL2 for the first method and 50 μg/mL PHA for the ...
Pitfalls in determining the cytokine profile of human T cells
2013年4月30日 · Two of the most widely applied methods for activation of T cells are phorbol 12-myristate 13-acetate (PMA) together with Ionomycin or anti-CD3/anti-CD28 stimulation. We analyzed production of IL-4, IFN-γ, IL-17 and IL-10 by a panel of human CD4 T-cell clones isolated from intestinal biopsies using the Bio-Plex™ assay and also flow-cytometry ...
The Effects of Stimulation with PMA/Ionomycin on CD4+ T Cell …
CD4+ T cell frequency and CD4 MFI (mean fluorescence intensity) were evaluated using flow cytometry before and after stimulation with PMA/ION. Results: The frequencies of CD4+ T cells were significantly lower in patients with LRBA deficiency than in HCs before and after treatment.
Ca2+ signals are essential for T-cell proliferation, while Zn2
2024年7月24日 · Phorbol 12-myristate 13-acetate (PMA) and calcium ionophores are commonly used for T-cell activation and PMA stimulation was already shown to increase intracellular Zn 2+ levels in Jurkat...
NFAT and NF-κB dynamically co-regulate TCR and CAR ... - Cell …
2023年7月25日 · Here, we show that two key pathways in T cell signaling, calcium/nuclear factor of activated T cells (NFAT) and protein kinase C (PKC)/nuclear factor κB (NF-κB), integrate through a dynamic and combinatorial strategy to fine-tune T cell response genes.