Comparison of the vector systems for gene transduction into rat dendritic cells and peritoneal exudate cells.
Specialized antigen-presenting cells (APC), known as dendritic cells (DC), play a pivotal role in initiating primary immune responses. It has been reported that several vector systems, including adenoviral vectors, retroviral vectors, Hemagglutinating Virus of Japan (HVJ)-related vectors, and electroporation, are able to transduce genes into mouse and human DC. This has not been achieved for rat DC. To our knowledge, there is no direct evidence to support the view that the currently used vector systems are able to transduce genes into mature DC. Because most, if not all, gene transfer studies investigating DC or DC-related cell populations are carried out using heterogeneous groups of cells, it is therefore very important to determine to what extent gene transduction occurs in rat DC, and also selected mature DC (CD161a+ fully mature DC). In this study, we provide evidence that none of 4 vector systems are able to transfer genes into fully mature rat DC, which are derived from bone marrow cells (BMC), driven by Flt3/Flk2 ligand and interleukin (IL)-6, and purified by CD161a. Nevertheless, the most efficient gene transduction was observed in the developing DC progenitor cells during the long-term culture of rat BMC, and its gene expression was successfully achieved after 2 weeks of culture only with a human immunodeficiency virus (HIV)-based lentiviral vector system. The most critical time point for lentiviral gene transduction was around the 7th day from the beginning of culture with lentiviral vectors. Rat peritoneal exudate cells (PEC) and another cell line (K562) were easily transducted by adenoviral vectors and lentiviral vectors.
Satoh
E
E
Department of Research Surgery, National Center for Child Health and Development, Ohkura, Tokyo, Japan.
Hara
Y
Y
Fuji
N
N
Li
X-K
XK
Teramoto
K
K
Arii
S
S
Kimura
H
H
D000256:Adenoviridae / Q000502:physiology*; D000818:Animals; D000938:Antigen-Presenting Cells / Q000502:physiology; D001854:Bone Marrow Cells / Q000502:physiology; D003713:Dendritic Cells / Q000502:physiology*; D018274:Electroporation; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D029082:Sendai virus / Q000235:genetics*; D014161:Transduction, Genetic
D049452:Green Fluorescent Proteins