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An expanding area of interest is the utilization of microbe-based components to augment mucosal and systemic immune responses to target antigens. Thus, the aim of the present study was to assess if the flagellin component FljB from Salmonella enterica serovar Typhimurium could act as a mucosal adjuvant and then to determine the cellular mechanism(s) by which FljB mediates its adjuvant properties. To determine if FljB could act as a mucosal adjuvant, mice were immunized by the intranasal (i.n.) route with antigen alone or in conjunction with FljB. Additionally, we assessed how FljB affected the levels of the costimulatory molecules B7-1 and B7-2 on dendritic cells by flow cytometry and determined the functional role these costimulatory molecules played in the adjuvant properties of FljB in vivo. Mice immunized by the i.n. route with antigen and FljB exhibited significantly elevated levels of mucosal and systemic antibody and CD4؉ -T-cell responses compared to mice given antigen only. Stimulation of dendritic cells in vitro with FljB resulted in a pronounced increase in the surface expression of B7-1 and B7-2. The percentage of dendritic cells expressing B7-2 but not B7-1 increased significantly when stimulated with FljB over a concentration range of 10 to 10,000 ng/ml. Immunization of wild-type and B7-1, B7-2, and B7-1/2 knockout mice by the i.n. route revealed that the ability of FljB to increase B7-2 expression is largely responsible for its adjuvant effect in vivo. These findings demonstrate that FljB can act as an effective mucosal adjuvant and that its ability to enhance the level of B7-2 expression is predominantly responsible for its adjuvant properties.

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