The change of immunoactivity of dendritic cells induced by mouse 4-1BBL recombinant adenovirus.
OBJECTIVE
The purpose of this study is to construct a recombinant adenovirus vector carrying mouse 4-1BBL and observe its effects in dendritic cells.


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-1 was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. After the co-culture of dendritic cells (DCs) with Ad-4-1BBL-transfected RM-1 cells, interleukin (IL)-6 and IL-12 production were assessed by enzyme-linked immunosorbent assay (ELISA) and co-stimulatary molecules (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.


CONCLUSIONS
The results indicated the recombinant mouse 4-1BBL can effectively activate DCs.
Youlin
Kuang
K
Department of Urology, Renmin Hospital of Wuhan University, Wuhan University, Wuhan 430060, China.
Xiaodong
Weng
W
Xiuheng
Liu
L
Zhiyuan
Chen
C
Hengcheng
Zhu
Z
Hui
Chen
C
Botao
Jiang
J
D053330:4-1BB Ligand / Q000235:genetics / Q000502:physiology*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D053330:4-1BB Ligand; D016207:Cytokines; D012333:RNA, Messenger; D011994:Recombinant Proteins