It is possible that the BBK32 homolog

in N40D10/E9 was significantly different from the BBK32 of B31 both at DNA and protein level, and hence, may not carry out the same functions. This can also explain a higher level of binding of B31 strain to Vero cells and potentially other cell lines that are not part of this study since in addition to its ability to recognize GAGs, BBK32 is also a fibronectin-binding protein [41, 53]. Interestingly, the N40 strain with published sequence is different from our N40D10/E9 clone. The sequenced N40 contains a bbk32 gene, which is similar to the bbk32 of B31 with 96% identity and 97% similarity with the B31 protein. In another study, we have shown that lp36, which contains the bbk32 gene in the B31 strain, is missing only in our N40 strain [29]. It is likely that the BBK32 protein, and potentially other unidentified

adhesins, may contribute to the binding of the B31 click here strain, and not of N40D10/E9. BBK32 may recognize fibronectin as a component of the extracellular matrix of the Vero cells. A selleckchem predicted higher pathogenicity of the B31 strain relative to the N40D10/E9 strain based-upon RST and ospC grouping contradicts both our results and the findings of other researchers, who have used N40 strains [23, 35, 105, 106]. Thus, RST and one virulence factor (ospC) sequence comparison may be important for phylogenetic analysis but may not be suitable for drawing conclusions about the pathogenicity of a particular strain of B. burgdorferi without assessment of the virulence factors or actually conducting the experiments. However, due to development BYL719 concentration of genetic manipulation techniques for B. burgdorferi only in the last decade, the roles of only a few virulence factors have been determined, and a comprehensive analysis Progesterone of multi-virulence loci of B31 and N40D10/E9 strains is not yet possible. Furthermore, a full repertoire of the virulence factors for Lyme spirochetes is still not determined even on the basis of the sequence homology with genes of other pathogens since spirochetes contain most unique genes [101, 107]. Finally, genetic manipulation and evaluation

of mutant B. burgdorferi strains remains very time consuming and difficult. Therefore, the pathogenicity of B31 and N40D10/E9 cannot be determined simply by multi-virulence locus sequence typing (MVLST) at present similar to that used for other pathogenic bacteria [5, 6, 108]. Therefore, we used an alternative approach to investigate the functions relevant to tissue colonization of B31 and N40D10/E9 strains in vitro and examined their virulence in the mouse model. Interestingly, even though it was possible to determine the molecular basis of adherence using the mammalian cell lines, we did not see a direct correlation of the ability of these strains to adhere to the mammalian cells in vitro and infectivity or pathogenicity in the mouse host.