Cells transfected with a non-neutralizing antibody gene are resistant to HIV infection: targeting the endoplasmic reticulum and trans-Golgi network.
Plasmids containing single chain Fv (scFv) non-neutralizing human anti-HIV-1 gp41 Ab cDNA, with or without endoplasmic reticulum (ER) or trans-Golgi network (TGN) retention signals, were constructed. Stable transfectants expressing these scFvs then were generated from COS-7 cells and HIV-1-susceptible CD4+ human T cells (Jurkat). scFv without a retention signal was secreted from cells, whereas scFv with an ER or TGN retention signal remained primarily within targeted intracellular compartments. The expression of scFv, scFv-ER, and scFv-TGN did not adversely affect the appearance of uninfected cells, as measured by growth rate or CD4 expression. Pulse-chase experiments revealed that the t(1/2) of scFv-ER and scFv-TGN within cells was greater than 24 h and less than 9 h, respectively. The scFv-ER and scFv-TGN bound HIV gp160, and the scFv-ER-gp160 and the scFv-TGN-gp160 complexes were stable within HIV-infected transfectants. Further studies revealed that the maturation processing of gp160 into gp120 and gp41 was blocked in the scFv-ER transfectants, but not in the scFv-TGN transfectants. Moreover, HIV replication, as measured by p24, was inhibited by up to 99% in cells transfected with scFv-ER or scFv-TGN, but was not inhibited in cells transfected with the secretory form of scFv. It is concluded that the targeting of non-neutralizing anti-HIV-1 Abs to specific intracellular compartments blocks HIV replication and represents a potential therapeutic strategy for protecting uninfected lymphopoietic stem cells from HIV-1-infected patients.
Zhou
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Oral Infection and Immunity Branch, National Institute of Dental Research, Bethesda, MD 20892, USA.
Goldstein
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S
Devadas
K
K
Tewari
D
D
Notkins
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D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D004721:Endoplasmic Reticulum / Q000276:immunology* / Q000378:metabolism; D015686:Gene Products, env / Q000378:metabolism; D015726:Giant Cells / Q000276:immunology / Q000821:virology; D006056:Golgi Apparatus / Q000276:immunology* / Q000378:metabolism; D019302:HIV Envelope Protein gp160 / Q000276:immunology; D015658:HIV Infections / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007113:Immunity, Innate; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D007424:Intracellular Fluid / Q000276:immunology / Q000378:metabolism; D019169:Jurkat Cells; D009500:Neutralization Tests; D011233:Precipitin Tests; D051381:Rats; D014162:Transfection / Q000235:genetics / Q000276:immunology*; D014779:Virus Replication / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D000998:Antiviral Agents; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv