Quantitative ELISPOT analysis of IFN-γ-producing lymphocytes in immunized mice after in vitro stimulation of isolated splenocytes (4 x 105cells) with the Brucella proteins L7/L12 or Omp16 or heat-inactivated B. abortus 19 (106 CFU). The mice were immunized with mono- and bivalent vaccine formulations of the subtypes Н5N1 or H1N1 by prime-boost intranasal (i.n.), conjunctival (c.) or subcutaneous (s.c.) vaccination 28 days apart, and with B. abortus 19 by single s.c. vaccination. Mice from the negative control groups were s.c. administered with PBS. Quantitative spot ± SE values (3 replicates per mouse) are presented for each group of 5 mice. *Р < 0.005 or Р < 0.0001 compared to the all group of animals vaccinated with the Flu-NS1-124-L7/L12 (Н5N1 + H1N1) viral constructs; **Р < 0.05 or Р < 0.025 compared with º; ***Р < 0.05 or Р < 0.025 compared with †. Statistical analysis was performed using a one way ANOVA followed by Tukey’s multiple comparisons test.