title: An ex vivo readout for evaluation of dendritic cell-induced autologous cytotoxic T lymphocyte responses against esophageal cancer.
abstract: Esophageal cancer is a highly malignant disease that despite surgery and adjuvant therapies has an extremely poor outcome. Dendritic cell (DC) immunotherapy as a novel promising strategy could be an alternative for treating this malignancy. Effective DC-mediated immune responses can be achieved by raising cytotoxic T lymphocyte (CTL) response against multiple antigens through loading DCs with total tumor RNA. However, the efficacy of this strategy first needs to be evaluated in a pre-clinical setting. The aim of the study was to set up an ex vivo autologous human readout assay for assessing the effects of DC-mediated cytotoxic responses, using total tumor RNA as an antigen load. Biopsy specimens of seven esophageal cancer patients were used to establish primary cultures of normal and cancer cells and to obtain autologous RNA for loading DCs. Mature DCs loaded with either normal or tumor RNA were obtained and subsequently used to raise various lymphocytes populations. Apoptosis levels of the autologous cultures were measured before and after incubating the cultures with the different lymphocytes populations. The mean apoptosis levels in the tumor cell cultures, induced by lymphocytes instructed by DCs loaded with tumor RNA, significantly increased with 15.6% +/-2.9 SEM (range 3.4-24.5%, t-test, P < 0.05). Incubation of the normal cultures with the lymphocytes populations showed a mean non-significant increase in apoptosis of 0.4% +/-3.4 SEM (range -13.9 to 9.8%, t-test, P = 0.7). Here, we introduce a practical, patient-specific autologous readout assay for pre-clinical testing of DC-mediated cytotoxic responses. Additionally, we demonstrated that the use of autologous tumor RNA as a strategy for raising cytotoxic responses against multiple tumor antigens could be effective for treating esophageal cancer.
authors.affiliation: Department of Experimental Internal Medicine, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. F.Milano@amc.uva.nl
authors.forename: Francesca
authors.initials: F
authors.lastname: Milano
authors.forename: Agnieszka M
authors.initials: AM
authors.lastname: Rygiel
authors.forename: Navtej
authors.initials: N
authors.lastname: Buttar
authors.forename: Jacques J G H M
authors.initials: JJ
authors.lastname: Bergman
authors.forename: Carine
authors.initials: C
authors.lastname: Sondermeijer
authors.forename: Jantine W P M
authors.initials: JW
authors.lastname: van Baal
authors.forename: Anja
authors.initials: A
authors.lastname: ten Brinke
authors.forename: Martien
authors.initials: M
authors.lastname: Kapsenberg
authors.forename: S Marieke
authors.initials: SM
authors.lastname: van Ham
authors.forename: Maikel P
authors.initials: MP
authors.lastname: Peppelenbosch
authors.forename: Kausilia K
authors.initials: KK
authors.lastname: Krishnadath
mesh_terms: D000230:Adenocarcinoma / Q000276:immunology / Q000628:therapy; D000368:Aged; D000951:Antigens, Neoplasm / Q000737:chemistry; D017209:Apoptosis; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology; D004938:Esophageal Neoplasms / Q000276:immunology* / Q000628:therapy*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*; D016896:Treatment Outcome
chemical_list: D000951:Antigens, Neoplasm