The Change of Immunoactivity of Dendritic Cells Induced by Mouse 4-1BBL Recombinant Adenovirus
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: The purpose of this study is to construct a recombinant adenovirus vector carrying mouse 4-1BBL and observe its effects in dendritic cells. Materials and Methods: Mouse 4-1BBL cDNA was taken from the plasmid pcDNA3-m4-1BBL and subcloned into adenovirus shuttle plasmid pAdTrack-CMV, and then transformed into competent BJ5183 with plasmid pAdEasy-1. After recombination in E. coli, Ad-4-1BBL was packaged and amplified in HEK 293 cells. The expression of 4-1BBL in Ad-4-1BBL-transfected mouse prostate cancer cell line RM-1was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. After the co-culture of dendritic cells (DCs) with Ad-4-1BBLtransfected RM-1 cells, interleukin (IL)-6 and IL-12 production were assessed by enzyme-linked immunosorbent assay (ELISA) and co-stimulatary moleculs (CD80 and CD86) on DCs were analyzed by flow cytometry. Results: The levels of IL-6 (3,960 pg/mL) and IL-12 (249 pg/mL) production in Ad-m4-1BBL-pulsed DCs were more than those in none-pulsed DCs. The differences were statistically significant (p < 0.05). The expression of co-stimulatary molecules (CD80 and CD86) was up-regulated in Ad-m4-1BBL-pulsed DCs. Conclusion: The results indicated the recombinant mouse 4-1BBL can effectively activate DCs.
Abstract
Purpose: The purpose of this study is to construct a recombinant adenovirus vector carrying mouse 4-1BBL and observe its effects in dendritic cells. Materials and Methods: Mouse 4-1BBL cDNA was taken from the plasmid pcDNA3-m4-1BBL and subcloned into adenovirus shuttle plasmid pAdTrack-CMV, and then transformed into competent BJ5183 with plasmid pAdEasy-1. After recombination in E. coli, Ad-4-1BBL was packaged and amplified in HEK 293 cells. The expression of 4-1BBL in Ad-4-1BBL-transfected mouse prostate cancer cell line RM-1was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. After the co-culture of dendritic cells (DCs) with Ad-4-1BBLtransfected RM-1 cells, interleukin (IL)-6 and IL-12 production were assessed by enzyme-linked immunosorbent assay (ELISA) and co-stimulatary moleculs (CD80 and CD86) on DCs were analyzed by flow cytometry. Results: The levels of IL-6 (3,960 pg/mL) and IL-12 (249 pg/mL) production in Ad-m4-1BBL-pulsed DCs were more than those in none-pulsed DCs. The differences were statistically significant (p < 0.05). The expression of co-stimulatary molecules (CD80 and CD86) was up-regulated in Ad-m4-1BBL-pulsed DCs. Conclusion: The results indicated the recombinant mouse 4-1BBL can effectively activate DCs.
참고문헌 (12)
- Nencioni A, "The use of dendritic cells in cancer immunotherapy", Crit Rev Oncol Hematol, 65, pp.191~199 (2008)
- Schwarz H, "Biological activities of reverse signal transduction through CD137 ligand", J Leukoc Biol, 77, pp.281~286 (2005)
- Laderach D, "4-1BB co-stimulation enhances human CD8(+) T cell priming by augmenting the proliferation and survival of effector CD8(+) T cells", Int Immunol, 14, pp.1155~1167 (2002)
- Lu ZY, "B7-1 and 4-1BB ligand expression on a myeloma cell line makes it possible to expand autologous tumor-specific cytotoxic T cells in vitro", Exp Hematol, 35, pp.443~453 (2007)
- Lee HW, "4-1BB promotes the survival of CD8+ T lymphocytes by increasing expression of Bcl-xL and Bfl-1", J Immunol, 169, pp.4882~4888 (2002)
- Croft M, "Co-stimulatory members of the TNFR family: keys to effective T-cell immunity?", Nat Rev Immunol, 3, pp.609~620 (2003)
- Kudo-Saito C, "4-1BB ligand enhances tumor-specific immunity of poxvirus vaccines", Vaccine, 24, pp.4975~4986 (2006)
- Xiao H, "Soluble PD-1 facilitates 4-1BBL-triggered antitumor immunity against murine H22 hepatocarcinoma in vivo", Clin Cancer Res, 13, pp.1823~1830 (2007)
- Futagawa T, "Expression and function of 4-1BB and 4-1BB ligand on murine dendritic cells", Int Immunol, 14, pp.275~286 (2002)
- Wiethe C, "Provision of 4-1BB ligand enhances effector and memory CTL responses generated by immunization with dendritic cells expressing a human tumor-associated antigen", J Immunol, 170, pp.2912~2922 (2003)
- Nencioni A, "The use of dendritic cells in cancer immunotherapy", Crit Rev Oncol Hematol, 65, pp.191~199 (2008)
- Hu Y, "In vitro antitumor cytotoxic T lymphocyte response induced by dendritic cells transduced with DeltaNp73alpha recombinant adenovirus", Oncol Rep, 18, pp.1085~1091 (2007)