title: RNA-electroporated CD40-activated B cells induce functional T-cell responses against HepG2 cells.
abstract: Hepatocellular carcinoma (HCC) is one of the incurable tumours in the world. Cell-based immunotherapy, in which antigen-loaded antigen-presenting cells (APCs) are able to elicit T cell responses, has become an alternative treatment for liver cancer. Here, we used HepG2 cells' total RNA-electroporated CD40 ligand-activated B (CD40-B) cells as alternative APC for induction of specific CD8+ T-cell responses. The antigen-presenting ability of CD40-B cells was determined by phenotypic analysis, showing a polyclonal, strongly activated B-cell population with high expression of co-stimulatory molecules. To demonstrate the ability of total RNA extracted from HepG2 cells electroporated CD40-B cells to induce CD8+ T-cell responses, these RNA-loaded cells were co-cultured with autologous peripheral blood mononuclear cells for 7 days followed by analysis of T-cell antigen specificity. These experiments showed that CD40-B cells electroporated with HepG2 cells' total RNA are capable of activating antigen-specific interferon-gamma-producing CD8+ T cells, and these T cells activated by CD40-B cells show a killing effect on HepG2 cells. These findings demonstrated that the carcinoma cell derived total RNA-electroporated CD40-B cells could be used as alternative APC for the induction of antigen-specific CD8+ T-cell responses, which might be used in HCC immunotherapy.
authors.affiliation: Department of Oncology, The Affiliated Drum Tower Hospital, Medical School of Naniing University and Clinical Cancer Institute of Nanjing University, Naniing, China.
authors.forename: S N
authors.initials: SN
authors.lastname: Shen
authors.forename: Z
authors.initials: Z
authors.lastname: Xu
authors.forename: X P
authors.initials: XP
authors.lastname: Qian
authors.forename: Y T
authors.initials: YT
authors.lastname: Ding
authors.forename: L X
authors.initials: LX
authors.lastname: Yu
authors.forename: B R
authors.initials: BR
authors.lastname: Liu
mesh_terms: D000328:Adult; D001402:B-Lymphocytes / Q000276:immunology*; D019013:CD40 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D018274:Electroporation / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008113:Liver Neoplasms / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D019013:CD40 Antigens; D012333:RNA, Messenger