Heterologous Expression of the Salmonella Enterica Serovar Paratyphi A Stk Fimbrial Operon Suggests a Potential for Repeat Sequence-mediated Low-frequency Phase Variation

Fimbriae mediate adhesion of Salmonella enterica organisms to the intestinal epithelium, which is an essential step in the pathogenesis process preceding invasion and/or systemic spread. In addition, Salmonella fimbrial genes transcripts were detected in the blood samples from Salmonella infected human patients, which supports the proposal that fimbriae play a role in invasive Salmonella infections. In this study, BlastN-based interrogation of the NCBI bacterial genome database and PCR investigation of Salmonella serovars have shown that the S. Paratyphi A stkF gene and/or the whole stk fimbrial gene cluster is present in about ~30% of S. enterica serovars investigated up to date. Furthermore, bioinformatics and phenotypic characterization have revealed that the stk fimbrial operon belongs to the chaperone/usher-γ4- fimbrial clade and that it encodes a mannose-sensitive hemagglutinating fimbrial structure. The latter trait is typical of type 1 fimbriae, in which fimbrial phase variation is common. The observed intragenic, 26 bp tandem repeat triplication event in stkF would suggest that slipped-strand mispairing and/or recombination within a signature stkF-borne tandem repeat motif as a likely mechanism for a form of low-frequency phase switching at the translational level leading to allelic OFF forms, hence the inability of production and/or absence of fimbriae by EM-examination on E. coli HB101/pUCstk-stkF. The in vitro profile of marked anti-StkF-mediated opsonophagocytosis and complement-mediated killing activity observed coupled with the mice immunogenicity profile strongly supports further investigation of StkF as a potential Salmonella vaccine candidate.