Vaccination via the pulmonary route could be an attractive alteative to parenteral administration.Research towards the best site of antigen deposition within the lungs to induce optimal immune responses has conflicting results which might be dependent on the type of vaccine and/or its physical state.Therefore,in this study,we explored whether deep lung deposition is crucial for two different vaccines,i.e.,influenza and hepatitis B vaccine.In view of this,influenza subunit vaccine and hepatitis B surface antigen were labeled with a fluorescent dye and then spray-dried.Imaging data showed that after pulmonary administration to mice the powders were deposited in the trachea/central airways when a commercially available insufflator was used while deep lung deposition was achieved when an in-house built aerosol generator was used.Immunogenicity studies revealed that comparable immune responses were induced upon trachea/central airways or deep lung targeting of dry influenza vaccine formulations.However,for hepatitis B vaccine,no immune responses were induced by trachea/central airways deposition whereas they were considerable after deep lung deposition.Thus,we conclude that deep lung targeting is not a critical parameter for the efficacy of pulmonary administered influenza vaccine whereas for hepatitis B vaccine it is.