Jpn. J. Infect. Dis., 57, 91-96, 2004
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Original Article
Down-Regulation of CXCR4 Expression in MT4 Cells by a Recombinant Vector Expressing Antisense RNA to CXCR4 and Its Potential Anti-HIV-1 Effect
Hui Chun Xing**, Xiao Yuan Xu, Zhen Liu***, Qin Huan Wang*, Min Yu and Chong Wen Si
Department of Infectious Diseases, Peking University First Hospital, Beijing 100034, P. R. China
(Received March 20, 2003. Accepted March 8, 2004)
*Corresponding author: Mailing address: No. 8, Xishiku Street,
Beijing 100034, P. R. China. Tel: +86-10-64437910, Fax: +86-10-66128182.
**Present address: Department of Infectious Diseases, First Hospital,
Zhejiang University, Hangzhou 310003, P. R. China.
***Present address: National Center for AIDS/STD Prevention and
Control, China CDC, Beijing 100034, P. R. China.
SUMMARY: CXC-chemokine receptor (CXCR4)
is one principle co-receptor for the entry of T cell line (T)-tropic
HIV-1 virus into a cell. In order to find more efficacious therapeutic
possibilities for@people with an HIV-1 infection, we explored
the inhibitory effects of antisense RNA on CXCR4
expression in MT4 cells. First, we used to RT-PCR to obtain DNA
fragments from healthy adult peripheral blood mononuclear cells;
these fragments targeted the initiation region of CXCR4
mRNA translation. We then constructed a recombinant retroviral
vector, pLXSN-X4a (containing antisense RNA to CXCR4).
After packaging by PA317 cells, the pseudovirion of the recombinant
vector had formed and succeeded in transfecting MT4 cells (a kind
of T-tropic HIV-1 susceptibility cell line). The PCR and RT-PCR
results showed that the recombinant vector had integrated into
the genome of MT4 cells and had been transcribed. The expression
of CXCR4 on the surface of MT4 cells transfected
with antisense RNA was reduced by 30%, compared with those cells
transfected with blank vector or untransfected cells. No change
in the DNA synthesis rates or in cell proliferation was found
in any of the transfected cells. After a challenge with HIV-1
SF33, the cells transfected with antisense RNA vector (pLXSN-X4a)
produced reduced p24 levels compared with the cells transfected
with blank vector (pLXSN) or untransfected cells. These results
indicated that these CXCR4-antisense expressing
cells could resist T-tropic HIV-1 infection and could retain normal
biological functions. These studies provide useful data for further
experiments in this area.
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