Female Balb/c mice (6-8 weeks) were divided of 3-4 mice for each group: control group, Squ/OVA, Spo-Squ/OVA, each of the above groups was administered intranasally at a dose of 25 μL free OVA(1 μg/μL) or OVA adjuvant(1:1) for one mouse; the other Squ/OVA subcutaneously at a dose of 25 μL for one mouse. Control group was PBS administered without vaccine.
The mice were immunized twice, once every two weeks. Blood samples were obtained from the retro-orbital plexus at day 28 after immunization. The blood could stand for 1 h after the clot was fully clotted. After centrifugation (HC-2062) (5000 r/min × 10 min), serum samples were preserved at -80°C and were eventually used for investigation of IgG and IgA by ELISA. Body weight was recorded daily for 28 days, Nasal washes and mucosal sections were taken from the nasal cavity of mice on day 28. Then the mice were sacrificed by cervical dislocation, and the nasal septum, spleen and thymus samples was surgically removed on day 28.
The mice were immunized twice, once every two weeks. Blood samples were obtained from the retro-orbital plexus at day 28 after immunization. The blood could stand for 1 h after the clot was fully clotted. After centrifugation (HC-2062) (5000 r/min × 10 min), serum samples were preserved at -80°C and were eventually used for investigation of IgG and IgA by ELISA. Body weight was recorded daily for 28 days, Nasal washes and mucosal sections were taken from the nasal cavity of mice on day 28. Then the mice were sacrificed by cervical dislocation, and the nasal septum, spleen and thymus samples was surgically removed on day 28.
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