Source: Hickey, J. W., et al. Enrich and Expand Rare Antigen-specific T Cells with Magnetic Nanoparticles. J. Vis. Exp. (2018).
This video describes a protocol for isolating rare-antigen-specific CD8+ T cells using magnetic nanoparticle-based artificial antigen-presenting cells (aAPCs) functionalized with antigenic peptide-loaded MHC-Igs and anti-CD28 antibodies. Interaction of the aAPCs with CD8+ T cells specifically expressing the antigenic peptide-targeting T cell receptors, followed by their expansion in vitro, results in the enrichment and expansion of this CD8+ T cell population.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Enrich Antigen-specific CD8+ T Cells with Prepared Nanoparticle Artificial Antigen-Presenting Cells.
2. Expand and Detect Antigen-specific CD8+ T Cells with Prepared Nanoparticle Artificial Antigen-Presenting Cells
The authors have nothing to disclose.
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DimerX I: Recombinant Soluble Dimeric Mouse H-2D[b]:Ig | BD Biosciences | 551323 | |
DimerX I: Recombinant Soluble Dimeric Mouse H-2K[b]:Ig Fusion Protein | BD Biosciences | 550750 | |
nanomag-D-spio, NH2, 100 nm nanoparticles | Micromod | 79-01-102 | |
96-Well Half-Area Microplate, black polystyrene | Corning | 3875 | |
Super Mag NHS-Activated Beads, 0.2 µm | Ocean Nanotech | SN0200 | |
Anti-Biotin MicroBeads UltraPure | Milteny | 130-105-637 | |
EZ-Link NHS-Biotin | ThermoFisher | 20217 | |
C57BL/6J (B6 wildtype) mice | Jackson Laboratory | 664 | |
CD8a+ T Cell Isolation Kit, Mouse | Miltenyi | 130-104-075 | |
MS Columns | Milteny | 130-042-201 | |
LS Columns | Milteny | 130-042-401 | |
Streptavidin-Phycoerythrin, SAvPE | Biolegend | 405203 | |
N52 disk magnets of 0.75 inches | K&J Magnetics | DX8C-N52 | |
APC anti-mouse CD8a Antibody, clone 53-6.7 | Biolegend | 100711 | |
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