Influenza infections are a major cause of mortality and morbidity worldwide. Therefore, there is a need to establish vaccines and immunization protocols that can prevent influenza infections. Herein, we show that one intranasal (IN) followed by one intramuscular (IM) immunizations with a combination of cell culture produced hemagglutinin (HA) antigens derived from 3 different influenza strains induced significantly higher serum hemagglutination inhibition (HI) and serum IgG antibody titers as well as T cell responses, compared to 2 IM, 2 IN or 1 M followed by 1 IN immunizations. Moreover, while 2 IM immunizations did not induce any antibody responses in nasal secretions or cervical lymph nodes, which drain the nasal mucosa, IN immunizations alone or in combination with IM immunization induced mucosal and local responses. These data show that the IN followed by IM immunization strategy holds promise to significantly raise serum and local antibody and T cell responses against seasonal influenza strains, and possibly pandemic influenza strains, for which no pre-existing immunity exists.