Resistance to human immunodeficiency virus type 1 (HIV-1) generated by lentivirus vector-mediated delivery of the CCR5Δ32 gene despite detectable expression of the HIV-1 co-receptors
It has previously been demonstrated that there are two distinct mechanisms for genetic resistance to human immunodeficiency virus type 1 (HIV-1) conferred by the CCR5Δ32 gene: the loss of wild-type CCR5 surface expression and the generation of CCR5Δ32 protein, which interacts with CXCR4. To analyse the protective effects of long-term expression of the CCR5Δ32 protein, recombinant lentiviral vectors were used to deliver the CCR5Δ32 gene into human cell lines and primary peripheral blood mononuclear cells that had been immortalized by human T-cell leukemia virus type 1. Blasticidin S-resistant cell lines expressing the lentivirus-encoded CCR5Δ32 showed a significant reduction in HIV-1 Env-mediated fusion assays. It was shown that CD4+ T lymphocytes expressing the lentivirus-encoded CCR5Δ32 gene were highly resistant to infection by a primary but not by a laboratory-adapted X4 strain, suggesting different infectivity requirements. In contrast to previous studies that analysed the CCR5Δ32 protective effects in a transient expression system, this study showed that long-term expression of CCR5Δ32 conferred resistance to HIV-1 despite cell-surface expression of the HIV co-receptors. The results suggest an additional unknown mechanism for generating the CCR5Δ32 resistance phenotype and support the hypothesis that the CCR5Δ32 protein acts as an HIV-suppressive factor by altering the stoichiometry of the molecules involved in HIV-1 entry. The lentiviral–CCR5Δ32 vectors offer a method of generating HIV-resistant cells by delivery of the CCR5Δ32 gene that may be useful for stem cell- or T-cell-based gene therapy for HIV-1 infection.
AgrawalL.,
LuX.,
QingwenJ.,
VanHorn-AliZ.,
NicolescueV.,
McDermottD.,
MurphyP. M.,
AlkhatibG.2004; Role for CCR5Δ32 protein in resistance to R5, R5X4, and X4 human immunodeficiency virus type 1 in primary CD4+ cells. J Virol 78:2277–2287[CrossRef]
AgrawalL.,
JinQ.,
AltenburgJ.,
MeyerL.,
TubianaR.,
TheodorouI.,
AlkhatibG.2007; CCR5Δ32 protein expression and stability are critical for resistance to human immunodeficiency virus type 1 in vivo . J Virol 81:8041–8049[CrossRef]
AlkhatibG.,
CombadiereC.,
BroderC. C.,
FengY.,
KennedyP. E.,
MurphyP. M.,
BergerE. A.1996; CC CKR5: a RANTES, MIP-1 α , MIP-1 β receptor as a fusion cofactor for macrophage-tropic HIV-1. Science 272:1955–1958[CrossRef]
BarryS. C.,
HarderB.,
BrzezinskiM.,
FlintL. Y.,
SeppenJ.,
OsborneW. R.2001; Lentivirus vectors encoding both central polypurine tract and posttranscriptional regulatory element provide enhanced transduction and transgene expression. Hum Gene Ther 12:1103–1108[CrossRef]
BleulC. C.,
WuL.,
HoxieJ. A.,
SpringerT. A.,
MackayC. R.1997; The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes. Proc Natl Acad Sci U S A 94:1925–1930[CrossRef]
BraciakT. A.,
BaconK.,
XingZ.,
TorryD. J.,
GrahamF. L.,
SchallT. J.,
RichardsC. D.,
CroitoruK.,
GauldieJ.1996; Overexpression of RANTES using a recombinant adenovirus vector induces the tissue-directed recruitment of monocytes to the lung. J Immunol 157:5076–5084
BroderC. C.,
BergerE. A.1995; Fusogenic selectivity of the envelope glycoprotein is a major determinant of human immunodeficiency virus type 1 tropism for CD4+ T-cell lines vs. primary macrophages. Proc Natl Acad Sci U S A 92:9004–9008[CrossRef]
BroxmeyerH. E.,
SrourE.,
OrschellC.,
IngramD. A.,
CooperS.,
PlettP. A.,
MeadL. E.,
YoderM. C.2006; Cord blood stem and progenitor cells. Methods Enzymol 419:439–473
ChelliM.,
AlizonM.2001; Determinants of the trans -dominant negative effect of truncated forms of the CCR5 chemokine receptor. J Biol Chem 276:46975–46982[CrossRef]
CollinsN. D.,
NewboundG. C.,
RatnerL.,
LairmoreM. D.1996; In vitro CD4+ lymphocyte transformation and infection in a rabbit model with a molecular clone of human T-cell lymphotrophic virus type 1. J Virol 70:7241–7246
DeanM.,
CarringtonM.,
WinklerC.,
HuttleyG. A.,
SmithM. W.,
AllikmetsR.,
GoedertJ. J.,
BuchbinderS. P.,
VittinghoffE.other authors1996; Genetic restriction of HIV-1 infection and progression to AIDS by a deletion allele of the CKR5 structural gene. Science 273:1856–1862[CrossRef]
DengH.,
LiuR.,
EllmeierW.,
ChoeS.,
UnutmazD.,
BurkhartM.,
Di MarzioP.,
MarmonS.,
SuttonR. E.other authors1996; Identification of a major co-receptor for primary isolates of HIV-1. Nature 381:661–666[CrossRef]
DragicT.,
LitwinV.,
AllawayG. P.,
MartinS. R.,
HuangY. X.,
NagashimaK. A.,
CayananC.,
MaddonP. J.,
KoupR. A.other authors1996; HIV-1 entry into CD4+ cells is mediated by the chemokine receptor CC-CKR-5. Nature 381:667–673[CrossRef]
HladikF.,
LiuH.,
SpeelmonE.,
Livingston-RosanoffD.,
WilsonS.,
SakchalathornP.,
HwangboY.,
GreeneB.,
ZhuT.,
McElrathM. J.2005; Combined effect of CCR5-Δ32 heterozygosity and the CCR5 promoter polymorphism −2459 A/G on CCR5 expression and resistance to human immunodeficiency virus type 1 transmission. J Virol 79:11677–11684[CrossRef]
KabatD.,
KozakS. L.,
WehrlyK.,
ChesebroB.1994; Differences in CD4 dependence for infectivity of laboratory-adapted and primary patient isolates of human immunodeficiency virus type 1. J Virol 68:2570–2577
KetasT. J.,
KuhmannS. E.,
PalmerA.,
ZuritaJ.,
HeW.,
AhujaS. K.,
KlasseP. J.,
MooreJ. P.2007; Cell surface expression of CCR5 and other host factors influence the inhibition of HIV-1 infection of human lymphocytes by CCR5 ligands. Virology 364:281–290[CrossRef]
KuhmannS. E.,
PlattE. J.,
KozakS. L.,
KabatD.2000; Cooperation of multiple CCR5 coreceptors is required for infections by human immunodeficiency virus type 1. J Virol 74:7005–7015[CrossRef]
LeeB.,
SharronM.,
MontanerL. J.,
WeissmanD.,
DomsR. W.1999; Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages. Proc Natl Acad Sci U S A 96:5215–5220[CrossRef]
MarkhamP. D.,
SalahuddinS. Z.,
KalyanaramanV. S.,
PopovicM.,
SarinP.,
GalloR. C.1983; Infection and transformation of fresh human umbilical cord blood cells by multiple sources of human T-cell leukemia-lymphoma virus (HTLV).. Int J Cancer 31:413–420[CrossRef]
MeyerL.,
MagierowskaM.,
HubertJ. B.,
RouziouxC.,
DeveauC.,
SansonF.,
DebreP.,
DelfraissyJ. F.,
TheodorouI.The SEROCO Study Group1997; Early protective effect of CCR-5Δ32 heterozygosity on HIV-1 disease progression: relationship with viral load. AIDS 11:F73–F78[CrossRef]
MiyoshiI.,
KubonishiI.,
YoshimotoS.,
AkagiT.,
OhtsukiY.,
ShiraishiY.,
NagataK.,
HinumaY.1981; Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature 294:770–771[CrossRef]
O'BrienS. J.,
MooreJ. P.2000; The effect of genetic variation in chemokines and their receptors on HIV transmission and progression to AIDS. Immunol Rev 177:99–111[CrossRef]
PopovicM.,
Lange-WantzinG.,
SarinP. S.,
MannD.,
GalloR. C.1983; Transformation of human umbilical cord blood T cells by human T-cell leukemia/lymphoma virus. Proc Natl Acad Sci U S A 80:5402–5406[CrossRef]
Rodríguez-FradeJ. M.,
del RealG.,
SerranoA.,
Hernanz-FalcónP.,
SorianoS. F.,
Vila-CoroA. J.,
de AnaA. M.,
LucasP.,
PrietoI.other authors2004; Blocking HIV-1 infection via CCR5 and CXCR4 receptors by acting in trans on the CCR2 chemokine receptor. EMBO J 23:66–76[CrossRef]
SunZ.,
DentonP. W.,
EstesJ. D.,
OthienoF. A.,
WeiB. L.,
WegeA. K.,
MelkusM. W.,
Padgett-ThomasA.,
ZupancicM.other authors2007; Intrarectal transmission, systemic infection, and CD4+ T cell depletion in humanized mice infected with HIV-1. J Exp Med 204:705–714[CrossRef]
VenkatesanS.,
PetrovicA.,
Van RykD. I.,
LocatiM.,
WeissmanD.,
MurphyP. M.2002; Reduced cell surface expression of CCR5 in CCR5Δ32 heterozygotes is mediated by gene dosage, rather than by receptor sequestration. J Biol Chem 277:2287–2301[CrossRef]
XiaoX.,
KinterA.,
BroderC. C.,
DimitrovD. S.2000; Interactions of CCR5 and CXCR4 with CD4 and gp120 in human blood monocyte-derived dendritic cells. Exp Mol Pathol 68:133–138[CrossRef]
YamamotoN.,
OkadaM.,
KoyanagiY.,
KannagiM.,
HinumaY.1982; Transformation of human leukocytes by cocultivation with an adult T cell leukemia virus producer cell line. Science 217:737–739[CrossRef]
ZimmermanP. A.,
Buckler-WhiteA.,
AlkhatibG.,
SpaldingT.,
KubofcikJ.,
CombadiereC.,
WeissmanD.,
CohenO.,
RubbertA.other authors1997; Inherited resistance to HIV-1 conferred by an inactivating mutation in CC chemokine receptor 5: studies in populations with contrasting clinical phenotypes, defined racial background, and quantified risk. Mol Med 3:23–36
Resistance to human immunodeficiency virus type 1 (HIV-1) generated by lentivirus vector-mediated delivery of the CCR5Δ32 gene despite detectable expression of the HIV-1 co-receptors