Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5

被引:571
作者
Qin, XF
An, DS
Chen, ISY
Baltimore, D
机构
[1] CALTECH, Div Biol, Pasadena, CA 91125 USA
[2] Univ Calif Los Angeles, David Geffen Sch Med, Dept Microbiol, Los Angeles, CA 90095 USA
[3] Univ Calif Los Angeles, David Geffen Sch Med, Dept Immunol & Mol Genet, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, David Geffen Sch Med, Dept Med, Los Angeles, CA 90095 USA
[5] Univ Calif Los Angeles, David Geffen Sch Med, AIDS Inst, Los Angeles, CA 90095 USA
关键词
D O I
10.1073/pnas.232688199
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Double-stranded RNAs approximate to21 nucleotides long [small interfering RNA (siRNA)] are recognized as powerful reagents to reduce the expression of specific genes. To use them as reagents to protect cells against viral infection, effective methods for introducing siRNAs into primary cells are required. Here, we describe success in constructing a lentivirus-based vector to introduce siRNAs against the HIV-1 coreceptor, CCR5, into human peripheral blood T lymphocytes. With high-titer vector stocks, >40% of the peripheral blood T lymphocytes could be transduced, and the expression of a potent CCR5-siRNA resulted in up to 10-fold inhibition of CCR5 expression on the cell surface over a period of 2 weeks in the absence of selection. In contrast, the expression of another major HIV-1 coreceptor, CXCR4, was not affected. Importantly, blocking CCR5 expression by siRNAs provided a substantial protection for the lymphocyte populations from CCR5-tropic HIV-1 virus infection, dropping infected cells by 3- to 7-fold; only a minimal effect on infection by a CXCR4-tropic virus was observed. Thus, our studies demonstrate the feasibility and potential of lentiviral vector-mediated delivery of siRNAs as a general means of intracellular immunization for the treatment of HIV-1 and other viral diseases.
引用
收藏
页码:183 / 188
页数:6
相关论文
共 61 条
[1]   Correction of ADA-SCID by stem cell gene therapy combined with nonmyeloablative conditioning [J].
Aiuti, A ;
Slavin, S ;
Aker, M ;
Ficara, F ;
Deola, S ;
Mortellaro, A ;
Morecki, S ;
Andolfi, G ;
Tabucchi, A ;
Carlucci, F ;
Marinello, E ;
Cattaneo, F ;
Vai, S ;
Servida, P ;
Miniero, R ;
Roncarolo, MG ;
Bordignon, C .
SCIENCE, 2002, 296 (5577) :2410-2413
[2]   CC CKRS: A RANTES, MIP-1 alpha, MIP-1 beta receptor as a fusion cofactor for macrophage-tropic HIV-1 [J].
Alkhatib, G ;
Combadiere, C ;
Broder, CC ;
Feng, Y ;
Kennedy, PE ;
Murphy, PM ;
Berger, EA .
SCIENCE, 1996, 272 (5270) :1955-1958
[3]   Marking and gene expression by a lentivirus vector in transplanted human and nonhuman primate CD34+ cells [J].
An, DS ;
Wersto, RP ;
Agricola, BA ;
Metzger, ME ;
Lu, S ;
Amado, RG ;
Chen, ISY ;
Donahue, RE .
JOURNAL OF VIROLOGY, 2000, 74 (03) :1286-1295
[4]   Lentivirus vector-mediated hematopoietic stem cell gene transfer of common gamma-chain cytokine receptor in rhesus macaques [J].
An, DS ;
Kung, SKP ;
Bonifacino, A ;
Wersto, RP ;
Metzger, ME ;
Agricola, BA ;
Mao, SH ;
Chen, ISY ;
Donahue, RE .
JOURNAL OF VIROLOGY, 2001, 75 (08) :3547-3555
[5]  
BALOTTA C, 1997, AIDS, V11, P67
[6]   GENE-THERAPY - INTRACELLULAR IMMUNIZATION [J].
BALTIMORE, D .
NATURE, 1988, 335 (6189) :395-396
[7]   Mechanism of transdominant inhibition of CCR5-mediated HIV-1 infection by ccr5Δ32 [J].
Benkirane, M ;
Jin, DY ;
Chun, RF ;
Koup, RA ;
Jeang, KT .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (49) :30603-30606
[8]  
BERGER EA, 1997, AIDS, V11, P3
[9]   HIV-1 infection in an individual homozygous for the CCR5 deletion allele [J].
Biti, R ;
French, RF ;
Young, J ;
Bennetts, B ;
Stewart, G ;
Liang, T .
NATURE MEDICINE, 1997, 3 (03) :252-253
[10]   The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes [J].
Bleul, CC ;
Wu, LJ ;
Hoxie, JA ;
Springer, TA ;
Mackay, CR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (05) :1925-1930