Analysis of the Epitopes Recognized by Mouse Monoclonal Antibodies Directed to Yersinia enterocolitica Heat‐Shock Protein 60

Abstract

To determine amino acid sequences of the epitopes recognized by monoclonal antibodies (mAbs) 3C8 and 5C3 directed against Yersinia enterocolitica heat-shock protein (HSP60), a dot blot analysis was perfomed using synthesized peptides of Y. enterocolitica HSP60 such as peptides p316-342, p327-359, p340-366, p316-326, p316-321, p319-323, and p321-326 which represent positions of amino acids in Y. enterocolitica HSP60. The dot blot analysis revealed that 5C3 mAb reacted with p316-342, p316-326 and p321-326, and 3C8 mAb p316-342 and p316-326. These results indicate that the epitopes recognized by the mAbs were associated with eleven amino acids, Asp Leu Gly Gln Ala Lys Arg Val Val Ile Asn, of p316-326. The sequence homology between p316-326 of Y. enterocolitica HSP60 and the rest of the HSP60 family suggests that the five amino acids of Lys, Arg, Val, Ile and Asn, which are highly conserved in the HSP60 family, might be related with the epitope recognized by 3C8. In contrast, it was also demonstrated that three amino acids of Leu, Gly and Val, which are not well conserved in the HSP60 family, might be related to the epitope recognized by 5C3.

Identification of Immunogenic Linear B-Cell Epitopes in C. burnetii Outer Membrane Proteins Using Immunoinformatics Approaches Reveals Potential Targets of Persistent Infections

“…From our point of view, the evaluation of the predicted epitopes' exposition in the protein oligomeric structure can be a critical step to improve the accuracy of epitope prediction, and the lack of those analyses could explain-at least in part-the low validation rates (15%) of predicted epitopes for some infectious agents seen in Brucella sp. proteins [81][82][83]. On the other hand, despite the improvements in methodologies and the results of in silico studies, the evaluation of reactivity to predicted linear B-cell epitopes remains rare, and this was the first study using samples from Brazilian C. burnetii-exposed individuals to confirm the natural immunogenicity of B-cell epitopes predicted in this bacterium's proteins.…”