A Vaccine-Pulsed Dendritic Cell-Based In Vitro Assay to Induce Lymphocyte Proliferation

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.