Human immunodeficiency virus infection and syncytium formation in HeLa cells expressing glycophospholipid-anchored CD4.
The CD4 molecule, a glycoprotein expressed primarily on the cell surface of specific T lymphocytes, is thought to function in T-cell antigen recognition and activation. In addition, CD4 serves as a receptor for human immunodeficiency virus type 1 (HIV-1) by a direct interaction with the HIV-1 surface glycoprotein (gp120). To further characterize the HIV-1-cell interaction, a HeLa cell line was established that expressed a chimeric molecule of CD4 and decay-accelerating factor (DAF). In the chimeric CD4-DAF molecule the transmembrane and cytoplasmic domains of CD4 were deleted and replaced with the carboxy-terminal 37 amino acids of DAF. This resulted in the anchoring of the extracellular domain of CD4 to the cell membrane via a glycophospholipid linkage. The glycophospholipid-anchored CD4 had a molecular size of approximately 56 to 62 kDa and was released following treatment of the cells with phosphatidylinositol-specific phospholipase C. HeLa cells expressing the CD4-DAF hybrid could be infected with HIV-1, as evidenced by reverse transcriptase activity, p24 core antigen content, and infectious virus production. In addition, transfection of the HeLa CD4-DAF cells with a plasmid that directs the synthesis of HIV-1 envelope glycoproteins or cocultivation with HeLa cells expressing the virus glycoproteins resulted in syncytium formation. These results indicate that the transmembrane and cytoplasmic domains of the CD4 molecule are dispensable for both HIV infection and syncytium formation.
Kost
T A
TA
Department of Molecular Biology, Glaxo Research Institute, Research Triangle Park, North Carolina 27709.
Kessler
J A
JA
Patel
I R
IR
Gray
J G
JG
Overton
L K
LK
Carter
S G
SG
D000163:Acquired Immunodeficiency Syndrome / Q000378:metabolism / Q000382:microbiology*; D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology; D018958:CD55 Antigens; D015683:Gene Products, gag / Q000276:immunology; D015726:Giant Cells / Q000276:immunology / Q000502:physiology*; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D006367:HeLa Cells / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014162:Transfection; D014758:Viral Core Proteins / Q000276:immunology; D014776:Virus Cultivation
D015704:CD4 Antigens; D018958:CD55 Antigens; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120; D008565:Membrane Proteins; D014758:Viral Core Proteins; D012194:RNA-Directed DNA Polymerase; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase