Cancer Immunology, Immunotherapy

, Volume 62, Issue 3, pp 571–583 | Cite as

Extensive expansion of primary human gamma delta T cells generates cytotoxic effector memory cells that can be labeled with Feraheme for cellular MRI

  • Gabrielle M. Siegers
  • Emeline J. Ribot
  • Armand Keating
  • Paula J. Foster
Original article

Abstract

Gamma delta T cells (GDTc) comprise a small subset of cytolytic T cells shown to kill malignant cells in vitro and in vivo. We have developed a novel protocol to expand GDTc from human blood whereby GDTc were initially expanded in the presence of alpha beta T cells (ABTc) that were then depleted prior to use. We achieved clinically relevant expansions of up to 18,485-fold total GDTc, with 18,849-fold expansion of the Vδ1 GDTc subset over 21 days. ABTc depletion yielded 88.1 ± 4.2 % GDTc purity, and GDTc continued to expand after separation. Immunophenotyping revealed that expanded GDTc were mostly CD27-CD45RA- and CD27-CD45RA+ effector memory cells. GDTc cytotoxicity against PC-3M prostate cancer, U87 glioblastoma and EM-2 leukemia cells was confirmed. Both expanded Vδ1 and Vδ2 GDTc were cytotoxic to PC-3M in a T cell antigen receptor- and CD18-dependent manner. We are the first to label GDTc with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles for cellular MRI. Using protamine sulfate and magnetofection, we achieved up to 40 % labeling with clinically approved Feraheme (Ferumoxytol), as determined by enumeration of Perls’ Prussian blue-stained cytospins. Electron microscopy at 2,800× magnification verified the presence of internalized clusters of iron oxide; however, high iron uptake correlated negatively with cell viability. We found improved USPIO uptake later in culture. MRI of GDTc in agarose phantoms was performed at 3 Tesla. The signal-to-noise ratios for unlabeled and labeled cells were 56 and 21, respectively. Thus, Feraheme-labeled GDTc could be readily detected in vitro via MRI.

Keywords

Gamma delta T cell expansion Gamma delta T cell cytotoxicity Iron labeling Preclinical cellular immunotherapy 

Supplementary material

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Gabrielle M. Siegers
    • 1
  • Emeline J. Ribot
    • 1
  • Armand Keating
    • 2
  • Paula J. Foster
    • 1
  1. 1.Imaging Research Laboratories, Robarts Research InstituteWestern UniversityLondonCanada
  2. 2.Cell Therapy Program, Princess Margaret HospitalUniversity of TorontoTorontoCanada

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