Home › Immunology › Killer Artificial Antigen Presenting Cells (KaAPC) for Efficient In Vitro Depletion of Human Antigen-specific T Cells
Immunology JoVE (Open Access) Citable · DOI

Killer Artificial Antigen Presenting Cells (KaAPC) for Efficient In Vitro Depletion of Human Antigen-specific T Cells

DOI: 10.3791/51859-v
What you'll learn
  • ✓ Generate HLA-A2-Ig fusion protein-based killer artificial antigen presenting cells
  • ✓ Perform quality control, peptide loading, and storage protocols for KaAPC
  • ✓ Execute in vitro CTL killing assays to measure antigen-specific T cell depletion
  • ✓ Evaluate antigen-specific elimination of cytotoxic T lymphocytes without systemic immune compromise
Protocol

Guidelines are presented for the generation of killer artificial antigen presenting cells, KaAPC, an efficient tool for in vitro depletion of human antigen-specific T cells and an alternative solution to cellular immunotherapy for the treatment of T cell-mediated autoimmune diseases in an antigen-specific fashion without compromising the remaining immune system.

Difficulty
advanced
Total time
~3–5 days (cell generation, quality control, peptide loading, killing assay)
Model organism
Human T cells (ex vivo); HEK293 or similar mammalian cell line for KaAPC production
Biosafety
BSL-2

Steps

1
Generate HLA-A2-Ig fusion protein KaAPC

Produce killer artificial antigen presenting cells using HLA-A2-Ig based constructs. This foundational step establishes the cell population used for subsequent depletion assays.

▶ 01:32
2
Perform quality control and peptide loading

Conduct quality control assessments, load KaAPC with cognate peptides, wash cells, and verify stability and storage conditions. Ensures reproducibility and functionality of the KaAPC preparation.

▶ 03:19
3
Execute KaAPC in vitro killing assay

Co-culture KaAPC with antigen-specific T cells under standardized conditions to measure cytotoxic elimination. Quantifies the depletion efficiency of target CTL populations.

▶ 04:53
4
Analyze antigen-specific CTL elimination results

Evaluate KaAPC-mediated killing of antigen-specific T cells while confirming preservation of non-cognate immune cells. Demonstrates selectivity and therapeutic potential of the approach.

▶ 06:23
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