Which cytokine activates dendritic cells?
Which cytokine activates dendritic cells?
Only rDCs and semi-mature dendritic cells express costimulatory factors CD86 and CD80 and secrete the cytokine IL-10 known to stimulate naïve Th0 cell differentiation into Th2 cells and Th3 or Tr1 regulatory T cells.
What role do dendritic cells play in the inflammatory response?
Dendritic cells (DCs) represent a heterogeneous family of immune cells that link innate and adaptive immunity. The main function of these innate cells is to capture, process, and present antigens to adaptive immune cells and mediate their polarization into effector cells (1).
Can you measure cytokines with flow cytometry?
Flow cytometry (FCM) is a highly effective technique that detects intracellular cytokines using specific fluorescence-labeled antibodies. The common steps of this assay include cell collection, fixation, permeabilization, blocking, intracellular staining and analysis by FCM.
What cells release cytokines?
Cytokines are mainly produced by macrophages and lymphocytes, although they can also be produced by polymorphonuclear leukocytes (PMN), endothelial and epithelial cells, adipocytes, and connective tissue. Cytokines are essential to the functions of macrophages.
How do dendritic cells get activated?
DCs are activated directly by conserved pathogen molecules and indirectly by inflammatory mediators produced by other cell types that recognise such molecules. In addition, it is likely that DCs are activated by poorly characterised cellular stress molecules and by disturbances in the internal milieu.
What is function of dendritic cells?
Dendritic cells (DCs) are antigen-presenting cells that capture, process, and present antigens to lymphocytes to initiate and regulate the adaptive immune response.
What is intracellular cytokine staining and how does it work?
Intracellular cytokine staining (ICS) of stimulated peripheral blood mononuclear cells (PBMC) followed by flow cytometric analysis is a well-established method for detecting immunological biomarkers in the form of expressed cytokines.
What factors affect the sensitivity of intracellular cytokine assays?
We have found that choices relating to the use of fresh whole blood or peripheral blood mononuclear cells (PBMC) and frozen PBMC; use of serum-containing or serum-free medium; length of stimulation period and use of co-stimulatory antibodies can all affect the sensitivity of intracellular cytokine assays.
How to determine frequency of cytokine production by flow cytometer?
Using intracellular cytokine staining method, the frequency of cytokine-producing cells and their phenotypes can be determined by flow cytometer. By using subset-specific antibody, we do not need to purify the T cells.
How do you measure cytokine secretion from T cells?
The capacity for a T cell to secrete certain cytokines can be measured using an intracellular cytokine staining protocol. Cells are harvested from the lungs and incubated with anti-CD3ε and anti-CD28 (both at 0.2 µg/10 6 cells) and monensin (3 µM) for 4 h at 37° C, 5% CO 2.
