Abstract
We used a monoclonal antibody (12G5) directed against an extracellular domain of CXCR-4 to investigate the role of this receptor in infection of immortalized lymphoid cell lines, peripheral blood mononuclear cells (PBMCs), and primary brain microglia with a dual-tropic strain of human immunodeficiency virus (HIV-1(89.6)) and a T-tropic strain (HIV-1(IIIB)). Addition of antibody 12G5 to cells prior to and during infection with HIV-1(89.6) inhibited p24 production 100- to 10,000-fold in CEMx174 and 174-CD4 cells and about 10-fold in PBMC cultures but had no activity against infection of either monocyte-derived macrophages or brain microglia. In contrast, 12G5 had little or no effect on infection of CEMx174 cells with HIV-1(IIIB) or HIV-1(HxB). To identify the region of the HIV-1(89.6) envelope that confers sensitivity to 12G5, we used chimeric molecular clones. Chimeras containing the V3 loop region of HIV-1(89.6) were inhibited by 12G5 to the same degree as wild-type HIV-1(89.6) whereas replication of those viruses containing the V3 loop of HIV-1(HxB) was not inhibited by the antibody. A similar pattern was seen in infections of a U87 glioblastoma line that coexpresses CD4 and CXCR-4. Antibody 12G5 was also able to block fusion between HeLa-CD4 cells and CEMx174 cells chronically infected with HIV-1(89.6) but had no effect on fusion mediated by cells chronically infected with HIV-1(IIIB). Taken together, these results suggest that different strains of HIV-1 may interact with different sites on CXCR-4 or may have different binding affinities for the coreceptor.
MeSH Terms
Antibodies, Monoclonal/metabolism
CD4 Antigens/metabolism
Cell Fusion
Cell Line
Cells, Cultured
HIV Core Protein p24/analysis
HIV Envelope Protein gp120/genetics,metabolism
HIV-1/isolation & purification,metabolism
HeLa Cells
Humans
Membrane Proteins/metabolism
Microglia/cytology,virology
Peptide Fragments/genetics,metabolism
Receptors, CXCR4
Receptors, HIV/metabolism
T-Lymphocytes/cytology,virology
Tumor Cells, Cultured
Chemicals
Antibodies, Monoclonal
CD4 Antigens
HIV Core Protein p24
HIV Envelope Protein gp120
HIV envelope protein gp120 (305-321)
Membrane Proteins
Peptide Fragments
Receptors, CXCR4
Receptors, HIV
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Strizki J M
Department of Neurology and Microbiology, University of Pennsylvania Medical Center, Philadelphia 19104-6146, USA.
Turner J D
Collman R G
Hoxie J
González-Scarano F
References (20)
20 references, click to expand
-
CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia.
Nature. 1997 Feb 13;385(6617):645-9
PMID: 9024664
-
Inhibition of human immunodeficiency virus fusion by a monoclonal antibody to a coreceptor (CXCR4) is both cell type and virus strain dependent.
J Virol. 1997 Feb;71(2):1692-6
PMID: 8995702
-
Replication of a macrophage-tropic strain of human immunodeficiency virus type 1 (HIV-1) in a hybrid cell line, CEMx174, suggests that cellular accessory molecules are required for HIV-1 entry.
J Virol. 1993 Nov;67(11):6707-15
PMID: 8411372
-
Cloning of a human seven-transmembrane domain receptor, LESTR, that is highly expressed in leukocytes.
J Biol Chem. 1994 Jan 7;269(1):232-7
PMID: 8276799
-
V3-independent determinants of macrophage tropism in a primary human immunodeficiency virus type 1 isolate.
J Virol. 1995 Mar;69(3):1755-61
PMID: 7853514
-
HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor.
Science. 1996 May 10;272(5263):872-7
PMID: 8629022
-
Identification of a major co-receptor for primary isolates of HIV-1.
Nature. 1996 Jun 20;381(6584):661-6
PMID: 8649511
-
The beta-chemokine receptors CCR3 and CCR5 facilitate infection by primary HIV-1 isolates.
Cell. 1996 Jun 28;85(7):1135-48
PMID: 8674119
-
A dual-tropic primary HIV-1 isolate that uses fusin and the beta-chemokine receptors CKR-5, CKR-3, and CKR-2b as fusion cofactors.
Cell. 1996 Jun 28;85(7):1149-58
PMID: 8674120
-
CC CKR5: a RANTES, MIP-1alpha, MIP-1beta receptor as a fusion cofactor for macrophage-tropic HIV-1.
Science. 1996 Jun 28;272(5270):1955-8
PMID: 8658171
-
A seven-transmembrane domain receptor involved in fusion and entry of T-cell-tropic human immunodeficiency virus type 1 strains.
J Virol. 1996 Sep;70(9):6288-95
PMID: 8709256
-
The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry.
Nature. 1996 Aug 29;382(6594):829-33
PMID: 8752280
-
Evidence for cell-surface association between fusin and the CD4-gp120 complex in human cell lines.
Science. 1996 Oct 25;274(5287):602-5
PMID: 8849450
-
A GTPase controlling nuclear trafficking: running the right way or walking RANdomly?
Cell. 1996 Oct 4;87(1):1-4
PMID: 8858141
-
Infection of primary human microglia and monocyte-derived macrophages with human immunodeficiency virus type 1 isolates: evidence of differential tropism.
J Virol. 1996 Nov;70(11):7654-62
PMID: 8892885
-
CD4-induced interaction of primary HIV-1 gp120 glycoproteins with the chemokine receptor CCR-5.
Nature. 1996 Nov 14;384(6605):179-83
PMID: 8906795
-
CD4-dependent, antibody-sensitive interactions between HIV-1 and its co-receptor CCR-5.
Nature. 1996 Nov 14;384(6605):184-7
PMID: 8906796
-
CD4-independent infection by HIV-2 is mediated by fusin/CXCR4.
Cell. 1996 Nov 15;87(4):745-56
PMID: 8929542
-
CD4, CXCR-4, and CCR-5 dependencies for infections by primary patient and laboratory-adapted isolates of human immunodeficiency virus type 1.
J Virol. 1997 Feb;71(2):873-82
PMID: 8995603
-
Molecular cloning of the cDNA and chromosomal localization of the gene for a putative seven-transmembrane segment (7-TMS) receptor isolated from human spleen.
Genomics. 1993 Jun;16(3):707-12
PMID: 8325644