Encyclopedia of Experiments: Immunology
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Transcript
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.
