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Expression of TAAs in glioblastoma and expansion of anti-TAA -reactive T cells

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Journal for ImmunoTherapy of Cancer20142 (Suppl 3) :P24

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  • Fresh Blood
  • Central Memory
  • Intracellular Cytokine
  • Protein Expression Pattern
  • Intracellular Cytokine Staining


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. TAAs expressed in glioblastoma may represent attractive targets for i) CARS, ii) transgenic T cells targeting nominal tumor antigens (e.g. NY-ESO-1) or iii) T cells enriched for TAA after ex vivo expansion.


Fresh blood samples were obtained from 50 patients with tumors of the central nervous system 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 by ELISA. PBMCs were expanded using IL-2/15/21 and peptides (15mers) covering TAAs (surviving or NY-ESO-1). The T cell phenotype (CD3, CD4, CD8, CD45RA and CCR7) was determined by flow cytometry and TAA-reactive T cells were identified by intracellular cytokine staining (IL-2, TNF, IFN and IL-17). TAA-specific IgG in serum was detected by a quantitative ELISA in patients with glioblastoma and in age- and sex-matched healthy donors. FGFRvIII was determined by RT-PCR and protein expression of survivin and NY-ESO-1 was evaluated by immunohistochemistry and graded using a scale from 1+ to 4+ along with pattern analysis of TAA expression.


We could detect IFN-gamma responses in 25% blood samples for NY-ESO-1 and 30% for survivin and antigen-specific CD8 / CD4+ T cell proliferation. Cellular responses could be augmented by adding cytokines, i.e. IL-2 and IL-7 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 ex vivo and exhibited TAA-specific production of IFNgamma and TNFalpha and a CD8+CD45RA-CCR7+ phenotype. 38/50 specimens expressed NY-ESO-1, yet only 3/50 showed a strong, universal (4+) NY-ESO-1 protein expression pattern, 12/50 cancer lesions exhibited a strong (4+) staining for survivin defined by immunohistochemistry. 25 % of glioblastoma tested positive for the FGFRvIII.


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 glioblastoma. TAA-reactive T cells can be successfully expanded from patients with glioblastoma in IL-2, IL-15 and IL-21, they exhibit a central memory phenotype and produce a Th1 cytokine cytokine pattern. NY-ESO-1 expression in glioblastoma represents a viable target for anti-NY-ESO-1 directed T cells.


Authors’ Affiliations

Dept. of Neurosurgery, Karolinska University Hospital, Stockholm, Sweden
Karolinska Insitutet, Stockholm, Sweden
Krankenhaus Nordwest, Frankfurt/M., Germany


© Dodoo et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.