Fig 1.
Screening for CD14 incorporation by lab-adapted HIV-1 isolates. (A) Schematic describing the virion capture assay used throughout. Briefly, antibodies or LPS (not depicted) were adsorbed onto 96-well plates and blocked before adding virus samples to allow precipitation of virus particles bearing the antigen of interest. Unbound virions were washed away and captured virions were lysed for quantitation of HIV-1 by Gagp24 ELISA. (B) Virion capture assay was performed on four HIV-1 isolates (IIIB, NL4-3, BaL, and SF162) produced by infected PBMCs using anti-CD14 (orange) and anti-gp120 (blue). Bars are isotype control-subtracted mean ± SD Gagp24 of duplicate assays and are representative of two independent experiments. (C) Western blot corroborating the incorporation of CD14. HIV-1 grown in monocyte-derived macrophages (MDMs) (BaL and SF162, two donors each) and PBMCs (IIIB, two donors; NL4-3, four donors) were concentrated, lysed, and resolved by SDS-PAGE before blotting for p24 (loading control), gp120 (positive control for viral surface protein), and CD14. Data are representative of three independent blots.