Use of a novel human immunodeficiency virus type 1 reporter virus expressing human placental alkaline phosphatase to detect an alternative viral receptor.
We report here on the construction and use of a novel human immunodeficiency virus (HIV) type 1 reporter vector, HIV-AP, that encodes human placental alkaline phosphatase. Upon staining with chromogenic alkaline phosphatase substrates 24 to 36 h postinfection, cells infected with HIV-AP develop an intense purple color and can then be counted under a dissecting microscope. Alternatively, HIV-AP infectivity can be quantitated and infected cells can be sorted by a fluorescence-activated cell sorter after staining with a fluorescent alkaline phosphatase substrate. The assay is rapid and accurate, has very low background in a variety of cell lines and primary cells, and is not restricted to use in human cells. Infectious HIV-AP can be pseudotyped by various HIV or murine leukemia virus envelope glycoproteins. Using this virus, we have addressed the long-standing question of CD4-independent infection of cells by HIV. Our results confirm the presence on a human osteosarcoma cell line of an alternative receptor for HIV infection that functions with an efficiency approximately 1/20 that of CD4.
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Aaron Diamond AIDS Research Center, New York University School of Medicine, New York 10016, USA.
Landau
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D000469:Alkaline Phosphatase / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015704:CD4 Antigens / Q000502:physiology; D005434:Flow Cytometry; D058851:GPI-Linked Proteins; D015686:Gene Products, env / Q000502:physiology; D017930:Genes, Reporter; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D015728:Receptors, HIV / Q000032:analysis*
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D058851:GPI-Linked Proteins; D015686:Gene Products, env; D007527:Isoenzymes; D015728:Receptors, HIV; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental