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Glycosyl Phosphatidylinositol-Anchored Variable Region of Llama Heavy Chain Only Antibody JM4 Efficiently Blocks Both Cell-Free and T Cell-T Cell Transmission of Human Immunodeficiency Virus Type 1.

Glycosyl Phosphatidylinositol-Anchored Variable Region of Llama Heavy Chain Only Antibody JM4 Efficiently Blocks Both Cell-Free and T Cell-T Cell Transmission of Human Immunodeficiency Virus Type 1.
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Liu L, Wang W, Matz J, Ye C, Bracq L, Delon J, Kimata JT, Chen Z, Benichou S, Zhou P,


Liu L, Wang W, Matz J, Ye C, Bracq L, Delon J, Kimata JT, Chen Z, Benichou S, Zhou P, (click to view)

Liu L, Wang W, Matz J, Ye C, Bracq L, Delon J, Kimata JT, Chen Z, Benichou S, Zhou P,

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Journal of virology 2016 9 21() pii

Abstract

JM2 and JM4 are two recently isolated variable regions (VHH) of heavy chain only antibodies from llamas that have been immunized with a trimeric gp140 bound to a CD4 mimic. JM2 binds the CD4-binding site of gp120 and neutralizes HIV-1 strains from subtypes B, C and G. JM4 binds gp120 and neutralizes HIV-1 strains from subtypes A, B, C, A/E and G in a CD4-dependent manner. In the present study, we constructed glycosyl-phosphatidylinositol (GPI)-anchored VHH JM2 and JM4 along with a E4 control and transduced them into human CD4(+) cell lines and primary CD4 T cells. We report that by genetically linking the VHHs with a GPI attachment signal, VHHs are targeted to the lipid rafts of the plasma membranes. Expression of GPI-VHH JM4, but not GPI-VHH E4 and JM2, on the surface of transduced TZM.bl cells potently neutralizes multiple subtypes of HIV-1 isolates including tier 2 or 3 strains, transmitted/founders, quasispecies and soluble single domain antibody (sdAb) JM4-resistant viruses. Moreover, transduction of CEMss-CCR5 cells with GPI-VHH JM4, but not with GPI-VHH E4, confers resistance to both cell-free and T cell-T cell transmission of HIV-1 and HIV-1 envelope-mediated fusion. Finally, GPI-VHH JM4-transduced human primary CD4 T cells efficiently resist both cell-free and T cell-T cell transmission of HIV-1. Thus, we conclude that the VHH JM4, when targeted to the lipid rafts of the plasma membrane, efficiently neutralizes HIV-1 infection via both cell-free and T cell-T cell transmission. Our findings should have important implications for GPI-anchored antibody-based therapy against HIV-1.

IMPORTANCE
Lipid rafts are specialized dynamic microdomains of the plasma membrane and have been shown to be gateways for HIV-1 budding as well as entry into T cells and macrophages. In nature, many glycosyl-phosphatidylinositol (GPI)-anchored proteins localize in the lipid rafts. In the present study, we developed GPI-anchored variable region (VHH) of two heavy chain only antibodies JM2 and JM4 from immunized llamas. We show that by genetically linking the VHHs with a GPI attachment signal, VHHs are targeted to the lipid rafts of the plasma membranes. GPI-VHH JM4, but not GPI-VHH JM2, in transduced CD4(+) cell lines and human primary CD4 T cells not only efficiently blocks diverse HIV-1 strains including tier 2 or 3 strains, transmitted founders, quasispecies and soluble sdAb JM4-resistant strains, but also efficiently interferes T cell-T cell transmissions of HIV-1 and HIV-1 envelope-mediated fusion. Our findings should have important implications in GPI-anchored antibody-based therapy against HIV-1.

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