Figure 4

Cytokine- and cell-mediated immune responses to LPAI H5N3 virus and soluble S. Gallinarum antigen. A The stimulation index (SI) in chicken PBMC samples stimulated with (1) H5N3 HA1 purified protein antigen and (2) soluble antigen from S. Gallinarum. The SI was calculated by dividing the mean OD value of antigen-stimulated wells by the mean OD of media-stimulated wells. B Cytokine levels in stimulated PBMCs from JOL1814 immunized and PBS control chickens. PBMCs from the 1814 oral, 1814 IM, vector and PBS control groups were collected at 11 weeks of age following booster immunization and stimulated with H5N3 HA1 purified protein antigen. Cytokine levels were assessed by qPCR with gene-specific primers, and the change was calculated using 2^−∆∆Ct. The fold change was calculated by taking the ct value for PBS control group as baseline. Each column represents the mean ± SD of six individual values. Oral: JOL1814 orally immunized group; IM: JOL1814 intramuscularly immunized group; VC: JOL1820 intramuscularly immunized group and PC: PBS inoculated group *p < 0.05; **p < 0.001 when compared with the PBS control group.