Immunization
Female Balb/c mice (6 weeks old , 18.1±1.4g) were randomly divided into five groups (n = 10 per group): normal, OVA, OVA+ M. alba L. 100 mg/kg, OVA+ M. alba L. 300 mg/kg, and OVA+ CT 0.5 mg/kg groups as described above. Mice were orally immunized four times at weekly intervals (days 0, 7, 14, and 21) with 10mg/kg of free OVA or OVA mixed with 100 or 300mg/kg/10mL of Morus alba L. dissolved in phosphate-buffered saline (PBS). The positive control mice were immunized with OVA mixed with 0.5mg/kg/5mL CT(cholea toxin) (Fig 1A).
Another female Balb/c mice (6 weeks old , 18.1±1.4g) were randomly divided into four groups (n = 10 per group): normal, control, vaccine, vaccine + M. alba L. 100 mg/kg, and OVA+ Alum 0.5 mg/kg groups as described above. Mice received H1N1 vaccine (12 HAU/100uL) administered twice, at a 0 and 14 days, in 100 𝜇L volume by intramuscular injection. The vaccine was diluted in saline to achieve the desired dose and mixed with alum (positive control) in a 1:1 ratio. The mice were orally immunized with 100 mg/kg Morus alba L. four times at weekly intervals (days 0, 7, 14, and 21) in PBS. The control mice were given PBS (Fig 1B).