Culturing of Activated T Cells from Human Peripheral Blood Mononuclear Cells
Transcrição
Multiple cell-surface receptor-ligand interactions between naïve T cells and antigen-presenting cells, APCs, elicit T cell activation and proliferation.
The two primary events in this process involve the recognition of the APC-expressed antigen by the CD3-T-cell receptor complex, followed by the interaction between the APC-expressed B7 ligand and the T cell receptor CD28.
To study T cell activation and proliferation in vitro, take human peripheral blood mononuclear cells, PBMCs, suspended in a suitable culture medium. Add magnetic beads coated with anti-CD3 and anti-CD28 antibodies, and divide the cells into two conical tubes.
Add the cytokine interleukin-2, IL-2, to the first tube and interleukin-15, IL-15, to the other. Transfer the cells to a multi-well plate. Incubate.
The beads' anti-CD3 and anti-CD28 antibodies bind to the CD3-T-cell receptor complex and CD28 receptor expressed on the T cells, respectively, causing T cell activation and proliferation.
With IL-2, the cytokine binds to its receptor on the activated T cell surface, leading to its differentiation and proliferation into effector T cells. Conversely, IL-15 primarily drives T cell differentiation and proliferation into memory T cells.
Resuspend and transfer half of the cells into new wells to allow cellular growth. Add fresh medium containing IL-2 or IL-15 to maintain the cytokine-differentiated T cell populations for subsequent analysis.
