Recombinant flagellins with partial deletions of the hypervariable domain lose antigenicity but not mucosal adjuvancy

Flagellin contains conserved N/C domains for TLR5 binding to activate innate immunity and a middle hypervariable domain harboring the major antigenic epitopes. However, conflict results existed in the previous studies as to whether the hypervariable domain was involved in the cytokine production and adjuvancy of flagellin. Here we constructed three flagellin variants (designated as FliC Delta 190-278, FliC Delta 220-320, and FliC Delta 180-400) with deletions in the hypervariable domain. Our data demonstrated that all deletion variants lost substantial antigenicity but not mucosal adjuvancy. Surprisingly, the variant with deletion of amino acids 220-320 (FliC Delta 220-320) induced higher production of IL-8, MCP-1, and TNF-alpha, and showed higher mucosal adjuvancy than full-length FliC flagellin. Our data supported the notion that the hypervariable domain was involved in the cytokine production by flagellin and more importantly demonstrated that the hypervariable domain was important for the mucosal adjuvancy of flagellin. (C) 2010 Elsevier Inc. All rights reserved.