[Preparation of recombinant adenovirus Ad5/F35 containing human WT1 and identification after recombinant adenovirus infected dendritic cells].
OBJECTIVE
To construct recombinant adenovirus vector Ad5/F35 containing human WT1 (Wilms' tumor gene-1) and observe its expression in the dendritic cells (DCs) derived from the peripheral blood of patients with melanoma.


METHODS
The two types of adenovirus vectors containing enhanced green fluorescence protein (EGFP), Ad5/F35-EGFP and Ad5-EGFP, were respectively infected into DCs. The EGFP expression was observed by fluorescent microscope. The adenovirus vector with the highest infection efficiency for the DCs was chosen to construct the recombinant adenoviral vector Ad5/F35-WT1, and WT1 expression in DCs was examined by immunohistochemistry and flow cytometry.


RESULTS
Ad5/F35 adenovirus was the vector with the better infection efficiency. The recombinant adenovirus vector Ad5/F35-WT1 was successfully constructed with the infective titer of 1×10(8); IU/mL, which was proved effective in mediating the expression of WT1 in the DCs by immunohistochemistry and flow cytometry.


CONCLUSIONS
The recombinant adenoviral vector Ad5/F35 containing human WT1 can transfer WT1 with effective expression into the DCs originally derived from the peripheral blood of patients with melanoma.
CTC Laboratory, Hematopoietic Stem Cell Transplantation Center, 307 Hospital of PLA, Beijing 100071; PLA Institute of Hematopoietic Stem Cell, Beijing 100071, China.
Xiaoying
X
Zhang
Department of Blood Transfusion, First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Yue
Y
Jin
CTC Laboratory, Hematopoietic Stem Cell Transplantation Center, 307 Hospital of PLA, Beijing 100071; PLA Institute of Hematopoietic Stem Cell, Beijing 100071, China.
Bin
B
Zhang
CTC Laboratory, Hematopoietic Stem Cell Transplantation Center, 307 Hospital of PLA, Beijing 100071; PLA Institute of Hematopoietic Stem Cell, Beijing 100071, China.
Hu
H
Chen
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology*; D004274:DNA, Recombinant / Q000235:genetics*; D003713:Dendritic Cells / Q000378:metabolism* / Q000821:virology*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007167:Immunotherapy; D025721:WT1 Proteins / Q000235:genetics*
D004274:DNA, Recombinant; D025721:WT1 Proteins