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T Cells Engineered to Express a T-Cell Receptor Specific for Glypican-3 to Recognize and Kill Hepatoma Cells In Vitro and in Mice
Christina Dargel
, Michal Bassani-Sternberg
, Julia Hasreiter
, Fabio Zani
, Jan-Hendrik Bockmann
, Frank Thiele
, Felix Bohne
, Karin Wisskirchen
, Susanne Wilde
, Martin F. Sprinzl
, Dolores J. Schendel
, Angela M. Krackhardt
, Wolfgang Uckert
, Dirk Wohlleber
, Matthias Schiemann
, Kerstin Stemmer
, Mathias Heikenwälder
, Dirk H. Busch
, Günther Richter
, Matthias Mann
, Ulrike Protzer correspondenceemail
Received: October 16, 2014; Accepted: May 30, 2015; Published Online: June 04, 2015
Article has an altmetric score of 4
DOI: http://dx.doi.org/10.1053/j.gastro.2015.05.055 |
Background & Aims
Cancer therapies are being developed based on our ability to direct T cells against tumor antigens. Glypican-3 (GPC3) is expressed by 75% of all hepatocellular carcinomas (HCC), but not in healthy liver tissue or other organs. We aimed to generate T cells with GPC3-specific receptors that recognize HCC and used them to eliminate GPC3-expressing xenograft tumors grown from human HCC cells in mice.
Methods
We used mass spectrometry to obtain a comprehensive peptidome from GPC3-expressing hepatoma cells after immune-affinity purification of human leukocyte antigen (HLA)-A2 and bioinformatics to identify immunodominant peptides. To circumvent GPC3 tolerance resulting from fetal expression, dendritic cells from HLA-A2−negative donors were cotransfected with GPC3 and HLA-A2 RNA to stimulate and expand antigen-specific T cells.
Results
Peptide GPC3367 was identified as a predominant peptide on HLA-A2. We used A2-GPC3367 multimers to detect, select for, and clone GPC3-specific T cells. These clones bound the A2-GPC3367 multimer and secreted interferon-γ when cultured with GPC3367, but not with control peptide-loaded cells. By genomic sequencing of these T-cell clones, we identified a gene encoding a dominant T-cell receptor. The gene was cloned and the sequence was codon optimized and expressed from a retroviral vector. Primary CD8+ T cells that expressed the transgenic T-cell receptor specifically bound GPC3367 on HLA-A2. These T cells killed GPC3-expressing hepatoma cells in culture and slowed growth of HCC xenograft tumors in mice.
Conclusions
We identified a GPC3367-specific T-cell receptor. Expression of this receptor by T cells allows them to recognize and kill GPC3-positive hepatoma cells. This finding could be used to advance development of adoptive T-cell therapy for HCC.
Keywords:
Cancer Immunotherapy, Tumor-Associated Antigens, Liver Cancer, Immune Response
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发表于 2015-9-24 17:42
