Selective Isolation of CD4+ Vδ1+ γδ T Cells from Human Peripheral Blood Mononuclear Cells

Published: August 31, 2023

Abstract

Source: Welker, C., et al. Isolation and Ex Vivo Culture of Vδ1+CD4+γδ T Cells, an Extrathymic αβT-cell Progenitor. J. Vis. Exp. (2015).

This video demonstrates a protocol for the magnetic labeling-based isolation of a low-concentration population of CD4+Vδ1+γδ T cells from human peripheral blood mononuclear cells (PBMCs). Using a combination of FITC-conjugated anti-Vδ1 antibodies, magnetic anti-FITC microbeads, and anti-CD4 antibody-coated superparamagnetic beads, the CD4+Vδ1+ T-cells can be successfully purified from the PBMC population.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Isolation of Vδ1 T Cells Take <1 x 106 isolated lymphocytes for an initial FACS determining the frequency of Vδ1 CD4 T cells. NOTE: The population size of this T-cell entity may differ greatly b…

Disclosures

The authors have nothing to disclose.

Materials

Phosphate-buffered Saline Sigma Aldrich D8537
MACS buffer Miltenyi Biotec 130-091-222 supplement with BSA and pre-cool before use
BSA Miltenyi Biotec 130-091-376 not mandatorily from this supplier
anti-human Vd1 FITC (clone: TS8.2) Thermo Scientific TCR2730 not mandatorily from this supplier
anti-human CD3 PerCP (clone: SK7) BD BioSciences 345766 not mandatorily from this supplier or this fluorochrome
anti-human TCRab PE (clone: T10B9.1A-31) BD BioSciences 555548 not mandatorily from this supplier or this fluorochrome
anti-human CD4 VioBlue (clone: MT466) Miltenyi Biotec 130-097-333 not mandatorily from this supplier or this fluorochrome
anti-human CD8 APC-H7 (clone: SK1) BD BioSciences 641400 not mandatorily from this supplier or this fluorochrome
Anti-FITC MultiSort Kit Miltenyi Biotec 130-058-701 yields better results than anti-FITC MicroBeads
MS columns Miltenyi Biotec 130-042-201 pre-cool before use
MiniMACS Separator Miltenyi Biotec 130-042-102
CD4 Positive Isolation Kit life technologies 11331D

Tags

Play Video

Cite This Article
Selective Isolation of CD4+ Vδ1+ γδ T Cells from Human Peripheral Blood Mononuclear Cells. J. Vis. Exp. (Pending Publication), e21563, doi: (2023).

View Video