Design and immunogenicity assessment of HIV-1 virus-like particles as a candidate vaccine

Zhang, XiZhen; Wang, XiaoDan; Zhao, DongHai; Meng, XiangYu; Zhao, XingHong; Yu, XiangHui; Kong, Wei

The plasmid pcDNA 3.1 EnvB/C and pNL4-3.Luc.R-E were cotransfected into 293T cells. Supernatants were collected 48 h after transfection incubated with blood at a final dilution of 1:1, 1:2, 1:4 and 1:8 for 1 h at 37°C. The mixture infected CD4-CXCR4-expressing HOS cells by DEAE. HOS cells were harvested, lysed, and tested after 72 h using the luciferase assay system (Promega) according to the manufacturer's instructions. The extent of luciferase expression was quantitated in a Dynex MLX luminometer and recorded in relative light units (r.l.u.). An inhibitory neutralization concentration, the concentration required to neutralize the preincubation mixture, was calculated by linear regression analysis.