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. 2015 Feb 14;15:58. doi: 10.1186/s12885-015-1044-0

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© Tazzari et al.; licensee BioMed Central. 2015

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 (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Phenotypic and functional analysis of tumor antigen-specific CD8 T cells. (A) Phenotypic analysis of pentamer⁺ CD8⁺ T cells after sensitization with the HLA2-A*0201 restricted-modified peptides (Melan-A/MART-1_[27L] or gp100_[210M]). (B) The tumor specificity of peptide sensitized T cells was assessed by measuring IFN-γ secretion (Enzyme-Linked ImmunoSpot (ELISpot) assay) upon stimulation with HLA-A*0201-restricted Melan A/MART-1 (modified or native)-pulsed (2 μmol/L) lymphoblastoid T2 cell line or HLA-matched HLA-A*0201⁺MART1⁺ tumor cells (#501mel and #624.38mel) pretreated or not with the anti-HLA class I (W6/32) mAb. Moreover, T cells were also incubated with HLA-mismatched allogeneic HLA-A*0201⁻MART1⁺ (#624.28mel) or HLA-A*0201⁺MART1⁻ melanoma cells (#A375mel). The irrelevant peptide NEF_[180–189] was used as negative control. P values were calculated by two-tailed t test.