Nanoparticle-Based Enrichment and Expansion of Antigen-Specific CD8+ T Cells

Published: August 31, 2023

Abstract

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.

Protocol

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. Isolate CD8+ T cells. Euthanize the animals by exposure to isoflurane followed by cervical dislocation. Remove the spleen and lymph nodes from wild-type C57BL/6j mice and place the…

開示

The authors have nothing to disclose.

Materials

DimerX I: Recombinant Soluble Dimeric Human HLA-A2:Ig Fusion Protein BD Biosciences 551263
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
LIVE/DEAD Fixable Green Dead Cell Stain Kit, for 488 nm excitation ThermoFisher L-34969

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記事を引用
Nanoparticle-Based Enrichment and Expansion of Antigen-Specific CD8+ T Cells. J. Vis. Exp. (Pending Publication), e21571, doi: (2023).

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