The peripheral blood lymphocyte proliferation assay (21 (link)) was performed using a modified CCK8 method. Briefly, T-lymphocytes were isolated using a Peripheral Blood Lymphocyte Separation Kit (Solarbio, Beijing, China) according to manufacturer’s instruction. After cell counting, 80 μL diluted cells in RPMI 1640 medium (Gibco) were seeded into a 96-well plate. To specifically stimulate the proliferation of T-lymphocytes, 20 ul CHN-YC virus (106.25 TCID50/100 ul) or purified recombinant truncated E protein (20 mg/mL) were added; for mock group, equal volume of PBS was added. After 36 h cell cultured in 37 °C, cell proliferation was detected using a Cell Counting Kit-8 (MCE, Shanghai, China) per manufacturer’s instruction.
The expression of IFN-γ and IL-4 measured by ELISA. Th1-type cytokine IFN-γ and Th2-type cytokine IL-4 in serum at 14 dpi were measured using commercial duck IFN-γ and IL-4 sandwich ELISA kits (mlbio, shanghai, China) following the manufacturer’s instruction.
The expression of IFN-γ and IL-4 measured by ELISA. Th1-type cytokine IFN-γ and Th2-type cytokine IL-4 in serum at 14 dpi were measured using commercial duck IFN-γ and IL-4 sandwich ELISA kits (mlbio, shanghai, China) following the manufacturer’s instruction.
Free full text: Click here
