Nucleotide sequence and T cell epitopes of a membrane protein of Francisella tularensis

Abstract

A 17-kDa major membrane protein of the facultative intracellular bacterium Francisella tularensis is recognized by T cells from individuals previously undergoing tularemia or tularemia vaccination. Here the nucleotide sequence of the encoding gene is presented. The A + T rich sequence deduced a protein of 149 amino acids with a predicted m.w. of 15,772. Evidence is presented suggesting that the 17-kDa protein is a lipoprotein. Also, homology was found to a peptidoglycan-associated lipoprotein of Escherichia coli. Thirteen overlapping synthetic peptides encompassing the entire deduced protein were tested on lymphocytes from F. tularensis-primed individuals. Three peptides induced a T cell response, as disclosed by lymphocyte proliferation and production of IL-2 and IFN-gamma. Out of five responding individuals, only one recognized more than one of the peptides. A murine mAb recognized an epitope separate from the T cell-reactive peptides. The identified T cell epitopes are presumably relevant to the host defense against F. tularensis, especially because IFN-gamma is known to enhance the antimicrobial activity of the host against intracellular bacteria.