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A fast assay to gauge for TAA-reactive T cells in PBMCS from patients with pancreatic cancer

  • 1,
  • 2,
  • 2,
  • 2,
  • 2,
  • 2,
  • 3,
  • 2 and
  • 2
Journal for ImmunoTherapy of Cancer20142 (Suppl 3) :P25

https://doi.org/10.1186/2051-1426-2-S3-P25

  • Published:

Keywords

  • Cell Proliferation
  • Blood Sample
  • Flow Cytometry
  • Pancreatic Cancer
  • Cellular Response

Purpose

Active cellular therapy (ACT) using ex-vivo expanded T cells from patients with cancer, obtained by apheresis, can represent a viable source for anti-cancer directed cellular therapy. We established a T cell expansion protocol using 2 rounds of re-stimulation with TAA peptides along with IL-2, IL-15 and IL-21. In order to gauge the ex-vivo cellular reactivity as well as the potential to successful expand antigen-specific T cells from patients with pancreatic cancer, we established a screening assay using whole-heparin blood, to gauge for TAA reactivity (NY-ESO-1, survivin and mesothelin) and control antigens (EBNA-1, EBNA-3, CMVpp65).

Methods

Fresh blood samples were obtained from 24 patients with pancreatic cancer and from 6 individuals with pre-malignant lesions and tested for anti-TAA reactivity. T cells were expanded without cytokines, with IL-2 and IL-7, or with IL-2, IL-15 and IL-21 and tested for CD4/8 expansion by flow cytometry and for IFN-gamma production. PBMCs were expanded by cytokines and TAA peptides. CD3, CD4, CD8, CD45RA and CCR7 was determined by flow cytometry and TAA-reactive T cells were identified by ICS (IL-2, TNF, IFN and IL-17).

Results

We could detect IFN-gamma responses in 90% (27 in 30) in blood samples for mesothelin, 55,3% (16 in 30) for survivin and 43,3% (13 in 30) for NY-ESO-1. Cellular responses could be augmented by adding cytokines, i.e. IL-2 and IL-7 could favored CD4+ T cell proliferation, IL-2, IL-15 and IL-21 favored CD8+ T cell proliferation. TAAs-reactive T cells could be successfully expanded in vitro and exhibited TAA-specific production of IFNgamma and TNFalpha and a CD8+CD45RA-CCR7+ phenotype.

Conclusion

A TAA-specific WBA (whole blood assay) can be used to gauge the potential for expansion of TAA-reactive T cells in peripheral blood from patients with pancreatic cancer. TAA-reactive T cells can be successfully expanded in IL-2, IL-15 and IL-21 and could represent a viable source for the cellular therapy of patients with pancreatic cancer.

Consent

Authors’ Affiliations

(1)
Karolinska Insitutet, Karolinska University Hospital, Stockholm, Sweden
(2)
Karolinska Insitutet, Stockholm, Sweden
(3)
Dept. of Neurosurgery, Karolinska University Hospital, Stockholm, Sweden

Copyright

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