Human immunodeficiency virus type 1 infection of SK-N-MC cells: domains of gp120 involved in entry into a CD4-negative, galactosyl ceramide/3' sulfo-galactosyl ceramide-positive cell line.

Abstract

The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl ceramide (GalCer) or a derivative molecule, 3' sulfogalactosyl ceramide (GalS), may serve as an alternative receptor for HIV type 1 (HIV-1) in cells of neural and colonic origin. Biochemical studies have demonstrated that recombinant gp120 envelope protein binds to GalCer/GalS in both solid-phase enzyme-linked immunosorbent assay and high-performance thin-layer chromatography overlays. We have used the SK-N-MC cell line, a CD4-negative, GalCer/GalS-positive cell line previously characterized as susceptible to HIV-1 infection, to identify virus isolates with either a positive infection phenotype, HIVHxB2, or a negative infection phenotype, HIV-1(89.6). Using a solid-phase virus binding assay, we determined the level of restriction in HIV-1(89.6) infection to be at the level of virus-glycolipid binding. Furthermore, using HIV-1HxB2-HIV-1(89.6) chimeras, we have identified a 193-amino-acid fragment from the envelope region of HIV-1HxB2 containing the V3, V4, and V5 regions which confers a positive infection phenotype on the HIV-1(89.6) background. Recombinant viruses which separate this 193-amino-acid fragment into two distinct chimeras are each able to confer a positive infection phenotype on the background of HIV89.6, suggesting that a stable GalCer/GalS-envelope interaction is dependent on the conformation of the envelope protein in the context of the viral membrane. Alternatively, the GalCer/GalS-gp120 bond may involve multiple sites on the oligomeric envelope protein.

• Authors: J M Harouse, R G Collman, F González-Scarano
• Journal: Journal of virology (J Virol) Vol. 69 Issue 12 Pg. 7383-90 (Dec 1995) ISSN: 0022-538X [Print] United States
• PMID: 7494242 (Publication Type: Comparative Study, Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antigens, CD
• CD4 Antigens
• DNA Primers
• DNA, Viral
• Galactosylceramides
• HIV Envelope Protein gp120
• Receptors, Virus
• Sulfoglycosphingolipids
• I(3)SO3-galactosylceramide

Topics

• Antigens, CD (genetics, physiology)
• Base Sequence
• Binding Sites
• CD4 Antigens (genetics, physiology)
• Cell Line
• Chromatography, High Pressure Liquid
• DNA Primers
• DNA, Viral (analysis, isolation & purification)
• Enzyme-Linked Immunosorbent Assay
• Galactosylceramides (analysis, physiology)
• HIV Envelope Protein gp120 (metabolism)
• HIV-1 (genetics, physiology)
• HeLa Cells
• Humans
• Kinetics
• Molecular Sequence Data
• Neuroblastoma
• Phenotype
• Polymerase Chain Reaction
• Receptors, Virus (analysis, physiology)
• Species Specificity
• Sulfoglycosphingolipids
• Tumor Cells, Cultured