title: Engineered cytotoxic T lymphocytes with AFP-specific TCR gene for adoptive immunotherapy in hepatocellular carcinoma.
abstract: Alpha-fetoprotein (AFP) is overexpressed in hepatocellular carcinoma (HCC) and could serve as a tumor-associated antigen (TAA) and potential target for adoptive immunotherapy. However, low frequency and severe functional impairment of AFP-specific T cells in vivo hamper adoptive infusion. TAA-specific T cell receptor (TCR) gene transfer could be an efficient and reliable alternation to generate AFP-specific cytotoxic T lymphocytes (CTLs). Autologous dendritic cells (DC) pulsed with AFP158-166 peptides were used to stimulate AFP-specific CTLs. TCR α/β chain genes of AFP-specific CTLs were cloned and linked by 2A peptide to form full-length TCR coding sequence synthesized into a lentiviral vector. Nonspecific activated T cells were engineered by lentivirus infection. Transgenetic CTLs were evaluated for transfection efficiency, expression of AFP158-166-specific TCR, interferon (IFN)-γ secretion, and specific cytotoxicity toward AFP+ HCC cells in vitro and in vivo. Flow cytometry revealed the AFP158-166-MHC-Pentamer positive transgenetic CTLs was 9.86 %. The number of IFN-γ secretion T cells and the specific cytotoxicity toward HpeG2 in vitro and in tumor-bearing NOD/SCID mice were significantly raised in transgenetic CTLs than that of AFP158-166-specific CTLs obtained by peptide-pulsed DCs or control group. TCR gene transfer is a promising strategy to generate AFP158-166-specific CTLs for the treatment of HCC.
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China.
authors.forename: Longhao
authors.initials: L
authors.lastname: Sun
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Guo
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China.
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Jiang
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China.
authors.forename: Li
authors.initials: L
authors.lastname: Lu
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.affiliation: Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road Heping District, Tianjin, 300052, China. xianghui_he001@163.com.
authors.forename: Xianghui
authors.initials: X
authors.lastname: He
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018014:Gene Transfer Techniques; D019672:Genes, T-Cell Receptor; D056945:Hep G2 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism*; D061986:MCF-7 Cells; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009000:Monocytes / Q000166:cytology; D010455:Peptides / Q000737:chemistry; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*; D019076:Transgenes; D000509:alpha-Fetoproteins / Q000378:metabolism*
chemical_list: D010455:Peptides; D011948:Receptors, Antigen, T-Cell; D000509:alpha-Fetoproteins; D007371:Interferon-gamma; D007770:L-Lactate Dehydrogenase
keywords: AFP158-166-specific T cell receptor; Adoptive immunotherapy; Hepatocellular carcinoma; Transgenetic