Figure 6.

ELISPOT assay and opsonophagocytosis killing assay. (A) Balb/c mice were immunized with the Kpn_Omp001, Kpn_Omp002, and Kpn_Omp005 individually or PBS, and mice were sacrificed 7 days after the second booster vaccination. After euthanasia, spleens were collected and a single suspension of splenocytes was obtained for cytokine-specific enzyme-linked Immunospot assay (ELISPOT assay), and the serum was isolated for the opsonophagocytosis killing assay. (B) Interferon gamma (IFN-γ), interleukin 4 (IL-4), and interleukin 17A (IL-17A)-producing splenocytes from vaccinated or control mice were analyzed using ELISPOT assay. SI: Immunized mice stimulated with KOMP, UI: Unstimulated immunized mice, SM: Mock mice stimulated with KOMP. (C) The opsonophagocytosis killing assay. The bacteria were incubated with heat-activated mouse antiserum against different KOMPs at 4°C for 20 min, differentiated HL-60 cells and rabbit complement co-incubated at 37°C for 1 h with agitation at 600 rpm, and samples were plated on BHI agar plates for CFU enumeration. **p < 0.01, ***p < 0.001.