title: A dendritic cell-based tumour vaccine for lung cancer: full-length XAGE-1b protein-pulsed dendritic cells induce specific cytotoxic T lymphocytes in vitro.
abstract: XAGE-1b is regarded as one of the most immunogenic antigens and the most promising targets for lung adenocarcinoma immunotherapy. In this study, we sought to determine whether monocyte-derived dendritic cells (DCs) pulsed with purified full-length XAGE-1b could induce specific cytotoxic T lymphocytes (CTLs) against tumour cells from patients with non-small cell lung cancer (NSCLC) in vitro. XAGE-1b mRNA expression was examined in primary cultures of lung cancer cells and normal lung epithelial cells established from fresh tissues surgically resected from 30 patients with NSCLC using reverse transcription-polymerase chain reaction (RT-PCR). XAGE-1b mRNA expression was observed in 11 of 18 (61.1%) adenocarcinomas and one of 12 (8.3%) lung cancers of other histological types (P = 0.015). The 246-base pairs XAGE-1b gene was inserted into a recombinant expression vector. Full-length XAGE-1b was then expressed in BL21 (DE3) Escherichia coli and purified by AKTA-fast performance liquid chromatography (FPLC). DCs generated from peripheral blood mononuclear cells were pulsed with XAGE-1b by incubation with the protein at an immature stage. The XAGE-1b-pulsed DCs induced CTLs following 14 days of co-culture. Finally, an adherent target detachment (ATD) assay was performed to test the cytotoxicity of the XAGE-1b-specific CTLs against cancer cells and normal lung epithelial cells. The XAGE-1b-specific CTLs had a stronger lytic effect on autologous XAGE-1b mRNA-positive cancer cells than on autologous XAGE-1b mRNA-negative cancer cells or allogenous XAGE-1b mRNA-positive cancer cells. The CTLs had no lytic activity against normal lung epithelial cells. These results can be used to develop simple and effective cancer/testis antigen-based immunotherapies for NSCLC.
authors.affiliation: Division of Pulmonary Oncology, Cancer Center and Lung Cancer Research Institute, Guangdong Provincial People's Hospital, Guangzhou, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Zhou
authors.forename: A-L
authors.initials: AL
authors.lastname: Guo
authors.forename: C-R
authors.initials: CR
authors.lastname: Xu
authors.forename: S-J
authors.initials: SJ
authors.lastname: An
authors.forename: Z
authors.initials: Z
authors.lastname: Wang
authors.forename: S-Q
authors.initials: SQ
authors.lastname: Yang
authors.forename: Y-L
authors.initials: YL
authors.lastname: Wu
mesh_terms: D000230:Adenocarcinoma / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology*; D002681:China; D003713:Dendritic Cells / Q000276:immunology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology*; D012333:RNA, Messenger / Q000276:immunology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism
chemical_list: D000951:Antigens, Neoplasm; D012333:RNA, Messenger; C413656:XAGE1A protein, human