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

Splenic HIV-1 antigen-specific T cells were identified indirectly using cytokine ELISPOT by measuring induced IFN- production (BD, Franklin Lakes, NJ, USA). Briefly, nitrocellulose bottomed 96-well plates were coated overnight with 5 g mL 1 anti-IFN- antibody at 4°C, and nonspecific binding was blocked for 1 h at 37°C. Purified splenocytes were dispensed in triplicate at a predetermined density. The plates were washed and the biotinylated detection antibody was added for another 1.5 h at 37°C. A 1/1000 dilution of horseradish peroxidase-streptavidin was added to the wells and incubated for 1 h, after which horseradish peroxidase-streptavidin substrate was added. After 30 min, the colorimetric reaction was terminated by washing with tap water. The spots were counted after drying.