An ex vivo readout for evaluation of dendritic cell-induced autologous cytotoxic T lymphocyte responses against esophageal cancer |
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Authors: | Francesca Milano Agnieszka M Rygiel Navtej Buttar Jacques J G H M Bergman Carine Sondermeijer Jantine W P M van Baal Anja ten Brinke Martien Kapsenberg S Marieke van Ham Maikel P Peppelenbosch Kausilia K Krishnadath |
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Institution: | (1) Department of Experimental Internal Medicine, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands;(2) Department of Gastroenterology, Mayo Clinic, Rochester, USA;(3) Department of Gastroenterology and Hepatology, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands;(4) Department of Cell Biology, Academic Medical Center, Amsterdam, The Netherlands;(5) Department of Immunopathology, Sanquin Research and Landsteiner Laboratory, Amsterdam, The Netherlands;(6) Department of Cell Biology and Histology, University Hospital, Groningen, The Netherlands |
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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.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Esophageal adenocarcinoma Dendritic cells Immunotherapy RNA electroporation Cytotoxic T-cells Apoptosis |
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