Down-regulation of CXCR4 expression by SDF-KDEL in CD34(+) hematopoietic stem cells: An anti-human immunodeficiency virus strategy.
CXCR4 plays an essential role as the first discovered coreceptor for the entry of T cell tropic isolates of HIV-1. Blocking the surface expression of this receptor may be a potential strategy to prevent HIV-1 infection. A lentiviral vector, pLenti6/V5-S-K, expressing a SDF-KDEL fusion protein was constructed and a replication-incompetent lentiviral stock was produced. The lentiviral stock was transduced into CD34(+) hHSC and the transient expression of the recombinant protein, SDF-1, was assayed using indirect immunofluorescence. The surface expression of CXCR4 in CD34(+) hHSC pretreated with different amounts of recombinant lentiviral vectors was detected by flow cytometric analysis. A marked down-regulation of CXCR4 expression in the cells transduced with recombinant lentiviral vectors pLenti6/V5-S-K was observed by flow cytometry with PE-conjugated anti-human CXCR4 monoclonal antibodies which showed the percentages of the inhibition effects of CXCR4-SDF-1 mediated syncytium formation are presented by concentration. P24 antigen levels of cell culture supernatants were detected on the 4th, 7th, and 10th day, with 10(3) TCID50 HIV-1 infected CD34(+) hHSC to evaluate the inhibitory effect of pLenti6/V5-S-K transduction on HIV-1 infection. The cells transfected with pLenti6/V5-S-K had a significant reduction of HIV-1 DP27 infection compared to controls (P<0.05).
Zhang
Jiu-Cong
JC
PLA Center for the Treatment of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, 1 XinSi Road, BaQiao District, Xi'an, China.
Sun
Li
L
Nie
Qing-He
QH
Huang
Chang-Xing
CX
Jia
Zhan-Sheng
ZS
Wang
Jiu-Ping
JP
Lian
Jian-Qi
JQ
Li
Xin-Hong
XH
Wang
Ping-Zhong
PZ
Zhang
Ye
Y
Zhuang
Yan
Y
Sun
Yong-Tao
YT
Bai
Xuefan
X
D018952:Antigens, CD34 / Q000032:analysis; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000096:biosynthesis* / Q000235:genetics; D003470:Culture Media / Q000737:chemistry; D015536:Down-Regulation; D005434:Flow Cytometry; D005822:Genetic Vectors; D016655:HIV Core Protein p24 / Q000032:analysis; D015497:HIV-1 / Q000254:growth & development*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000821:virology*; D006801:Humans; D007231:Infant, Newborn; D016086:Lentivirus / Q000235:genetics; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors*; D014161:Transduction, Genetic
D018952:Antigens, CD34; C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; D003470:Culture Media; D016655:HIV Core Protein p24; D019718:Receptors, CXCR4