A Vaccine-Pulsed Dendritic Cell-Based In Vitro Assay to Induce Lymphocyte Proliferation
筆記録
Begin by adding the test vaccine antigens to the dendritic cell culture.
The dendritic cell recognizes the antigen, internalizes it via receptor-mediated endocytosis, fragments it into smaller peptides, and transfers the resulting antigen-MHC complex to the cell surface for presentation.
Co-culture antigen-presenting dendritic cells with naïve T lymphocytes to enable interaction between the T cell receptor and the antigen-MHC complex, along with their co-receptors.
This leads to immunological synapse formation, activating T lymphocytes via intracellular signaling.
Activated T lymphocytes undergo clonal expansion, generating effector T lymphocytes.
Add interleukin-2, or IL-2, cytokine molecules to the culture and incubate. The binding of IL-2 triggers intracellular signaling, enabling the proliferation and survival of lymphocytes.
Take a small volume of the culture and centrifuge. Discard the supernatant. Resuspend the cell pellet with antigen-presenting dendritic cells and transfer it to the same well.
Restimulation with antigen increases lymphocyte proliferation.
Perform downstream analysis to quantify specific markers associated with lymphocyte proliferation.
