Journal: Vaccines
Article Title: A Pool of Ferritin Nanoparticles Delivering Six Proteins of African Swine Fever Virus Induces Robust Humoral and Cellular Immune Responses in Pigs
doi: 10.3390/vaccines14010093
Figure Lengend Snippet: Evaluation of the immunogenicity of the antigen candidates in pigs. ( A ) Schematic diagram of pig vaccination. Each pig was vaccinated with 100 μg of antigens at days 0, 14, and 28. Blood was collected at 0, 14, 28, and 42 days post-vaccination (dpv). ( B ) Verification of eukaryotic expression plasmids by Indirect immunofluorescence assay (IFA). HEK293T cells were transfected with plasmid pCAGGS-p30, pCAGGS-p54, pCAGGS-pE120R, pCAGGS-pH124R, pCAGGS-pE184L or pCAGGS-CD2v. At 24 h post-transfection, the cells were lysed with RIPA buffer and the whole cell lysates were verified by Western blotting with an anti-Flag monoclonal antibody. ( C ) IFA for ASFV antigen-specific antibody detections. By detecting the sera at 42 dpv to determine whether the antibody turned positive. Sera collected at 0 dpv were the negative control, and anti-Flag antibody was used as the positive control. ( D ) Antigen-specific antibody titers were determined by in-house indirect enzyme-linked immunosorbent assay (iELISA) for p30, p54, pE120R, pH124R, pE184L, and CD2v at 0 and 42 dpv. The dashed lines represent the cut-off value for each iELISA. Group 1: pigs vaccinated with p30 (1-1, 1-2 and 1-3; n = 3); Group 2: pigs vaccinated with p54 (2-1, 2-2 and 2-3; n = 3); Group 3: pigs vaccinated with pE120R (3-1, 3-2 and 3-3; n = 3); Group 4: pigs vaccinated with pH124R (4-1 and 4-2; n = 2); Group 5: pigs vaccinated with pE184L (5-1 and 5-2; n = 2); Group 6: pigs vaccinated with CD2v (6-3; n = 1). The reduced sample sizes in Groups 4-6 were due to unintended infection, and data from these groups should be interpreted as indicating immunological trends rather than definitive outcomes.
Article Snippet: Human embryonic kidney-derived HEK293T cells (Procell, Wuhan, China) were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Carlsbad, CA, USA) supplemented with 10% FBS.
Techniques: Immunopeptidomics, Expressing, Immunofluorescence, Transfection, Plasmid Preparation, Western Blot, Negative Control, Positive Control, Indirect ELISA, Infection
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