Abstract
Objective To investigate the ability of inducing antigen-specific cytotoxic T lymphocytes (CTL) stimulated by dendritic cell (DC) fused with MiaPaCa-2 cells in vitro. Methods DC were isolated and cultured from peripheral blood mononuclear cells (PBMCs). 50% PEG and 10% DMSO were used to fuse MiaPaCa-2 cells and DC, and DC co-cultured with MiaPaCa-2 cells and DC alone served as control. The fusion efficiency was assessed by flow cytometry (FCM) and DC-MiaPaCa-2 hybrids were identified as PE-MUC1/FITC- CD86 double positive cells. The survival rate of DC was determined by MTT method. The lymphocyte proliferation stimulated by DC in vitro was evaluated by mixed cell culture with DC in different ratios of 1∶10, 1∶20 and 1∶80. IL-2, IL-10, granzyme B and IFN-γ released by antigen-specific CTLs were measured by ELISA assay. Results The fusion rate in DC fused with MiaPaCa-2 cells (fused cells) was (42.30±7.30)%, which was higher than (7.21±1.06)% in DC co-cultured with MiaPaCa-2 cells(co-cultured cells). The cell viability of DC, co-cultured cells and fused cells was >95.0%, 85.0% and 62.8%, and fused cells had greatly lower cell viability than DC and co-cultured cells (P 0.05). At the co-culture ratio of 1∶10, IL-2 secreted by CTL in DC, co-cultured and fused cells was(27.30±5.21 ), (897.44±93.05), (2 243.80±381.46)ng/L; IL-10 was (19.55±2.05), (424.60±108.25), (706.53±161.29)ng/L; Granzyme B was(16.23±1.23), (451.07±120.50), (1327.77±205.15)ng/L; IFN-γ was (30.11±4.32 )、(982.00±124.68)、(2421.04±488.50)ng/L. Cytokines from the antigen-specific CTL induced by DC fused with MiaPaCa-2 cells were significantly higher than those by DC and DC co-cultured with MiaPaCa-2 cells ( all P<0.05). Conclusions The fusion of DC and pancreatic cancer MiaPaCa-2 cells could stimulate tumor antigen-specific CTL in vitro. Key words: Pancreatic neoplasms; Dendritic cells; Cell fusion; T-lymphocytes, cytotoxic
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
