Nanoparticle orientationally displayed antigen epitopes improve neutralizing antibody level in a model of porcine circovirus type 2

Ding, Peiyang; Zhang, Teng; Li, Yafei; Teng, Man; Sun, Yaning; Liu, Xiao; Chai, Shujun; Zhou, Enmin; Jin, Qianyue; Zhang, Gaiping

Nanoparticles orientationally display neutralizing epitopes cytokine production measurement RAW264.7 cells were first primed with 50 ng/mL LPS for 4 h, which increased the expression of pro-interleukin (IL)-1β and pro-IL-18, the immature states of IL-1β and IL-18, in cells, respectively. 32 After priming, the cells were seeded at 5×10 4 cells/well on 96-well cell culture plate. One hundred μL of AuNPs (10 μg/mL), Cap (1.2 μg/mL), or Cap-AuNPs (11.2 μg/mL) was added to incubate with the cells. Cap group and Cap-AuNPs group contain the same amount of Cap protein. Aluminum hydroxide was added as positive control. For IL-12 and tumor necrosis factor alpha (TNF-α) measurements, RAW264.7 cells were not primed with LPS. CpG was added as positive control. 33 After incubation for 24 h, cytokine concentration of IL-1β, IL-12, IL-18, and TNF-α was measured by ELISA following the manufacturer's protocols (R&D Systems, Minneapolis, MN, USA).