title: High mannose glycans and sialic acid on gp120 regulate binding of mannose-binding lectin (MBL) to HIV type 1.
abstract: Mannose-binding lectin (MBL) is a C-type lectin of the innate immune system that binds to carbohydrates on the surface of certain microorganisms. Previous studies showed that MBL binds to gp120, the envelope glycoprotein of HIV-1. gp120 is extensively glycosylated, with N-linked complex and high mannose carbohydrates accounting for about half of the molecular weight. The objectives of this study were to determine the types of glycans on gp120 important for MBL binding and to determine if alteration of complex glycans with neuraminidase (NA) could enhance the interaction of MBL with virus. Lectin blot analyses revealed that MBL interacted with recombinant gp120 (rgp120) from both T cell-tropic and M-tropic virus strains. Treatment of rgp120 with endoglycosidase H (eH) or endoglycosidase F1 (eF1) abrogated binding of MBL, but did not decrease binding of wheat germ agglutinin indicating that high mannose and/or hybrid N-linked glycans were required for MBL binding. Removal of sialic acids from rgp120 with NA enhanced MBL binding. Treatment of intact virus from T cell lines or primary isolates with eF1 also significantly decreased HIV binding to MBL, while treatment with NA substantially increased binding. Treatment of virus with both eF1 and NA did not decrease binding compared to NA alone suggesting that NA treatment exposed binding sites on gp120 that are not high mannose glycans. These studies provide evidence that MBL binds to HIV via high mannose carbohydrates on gp120 and shows that the interaction of MBL with virus is regulated by sialylation.
authors.affiliation: Rush-Presbyterian-St. Luke's Medical Center, Department of Immunology/Microbiology, Chicago, Illinois 60612, USA.
authors.forename: Melanie L
authors.initials: ML
authors.lastname: Hart
authors.forename: Mohammed
authors.initials: M
authors.lastname: Saifuddin
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Uemura
authors.forename: Eric G
authors.initials: EG
authors.lastname: Bremer
authors.forename: Brad
authors.initials: B
authors.lastname: Hooker
authors.forename: Toshisuke
authors.initials: T
authors.lastname: Kawasaki
authors.forename: Gregory T
authors.initials: GT
authors.lastname: Spear
mesh_terms: D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000378:metabolism*; D008358:Mannose / Q000502:physiology*; D037601:Mannose-Binding Lectin / Q000378:metabolism*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000494:pharmacology; D019158:N-Acetylneuraminic Acid / Q000502:physiology*; D009439:Neuraminidase / Q000494:pharmacology; D011134:Polysaccharides / Q000502:physiology*
chemical_list: D015699:HIV Envelope Protein gp120; D037601:Mannose-Binding Lectin; D011134:Polysaccharides; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D019158:N-Acetylneuraminic Acid; D008358:Mannose