Phage Display Detection of Mimotopes that Are Shared Epitopes of Clinically and Epidemiologically Relevant Enterobacteria

Armando Navarro,Alejandro Monsalvo-Reyes,Delia Licona-Moreno,Carlos A Eslava-Campos,Ulises Hernández-Chiñas

doi:10.3390/microorganisms8050780

Abstract

Background: Escherichia coli and Salmonella are etiologic agents of intestinal infections. A previous study showed the presence of shared epitopes between lipopolysaccharides (LPSs) of E. coli O157 and Salmonella. Aim: Using phage display, the aim of this study is to identify mimotopes of shared epitopes in different enterobacterial LPSs. Methods: We use anti-LPS IgG from E. coli O157 and Salmonella to select peptide mimotopes of the M13 phage. The amino acid sequence of the mimotopes is used to synthesize peptides, which are in turn used to immunize rabbits. The antibody response of the resulting sera against the LPSs and synthetic peptides (SPs) is analyzed by ELISA and by Western blot assays, indicating that LPS sites are recognized by the same antibody. In a complementary test, the reactions of human serum samples obtained from the general population against the SPs and LPSs are also analyzed. Results: From the last biopanning phase, sixty phagotopes are selected. The analysis of the peptide mimotope amino acid sequences shows that in 4 of them the S/N/A/PF motif is a common sequence. Antibodies from the sera of immunized rabbits with SP287/3, SP459/1, SP308/3, and SP073/14 react against both their own peptide and the different LPSs. The Western blot test shows a sera reaction against both the lateral chains and the cores of the LPSs. The analysis of the human sera shows a response against the SPs and LPSs. Conclusion: The designed synthetic peptides are mimotopes of LPS epitopes of Salmonella and E. coli that possess immunogenic capacity. These mimotopes could be considered for use in the design of vaccines against both enterobacteria.

Highlights

Diarrheal illnesses are an important public health problem around the world, accounting for more than 2 million deaths each year in children under 5 years of age, with those in developing countries being the most affected [1,2]
Enterobacteria are characterized by the cytoplasmic internal membrane, peptidoglycan, and the cell wall or external membrane, which is composed of lipopolysaccharides (LPSs) and different proteins [5]
The O antigen is a region that provides a hydrophilic surface to the bacterial cell wall and is composed of lateral carbohydrate chains that vary in their composition

Summary

Introduction

Diarrheal illnesses are an important public health problem around the world, accounting for more than 2 million deaths each year in children under 5 years of age, with those in developing countries being the most affected [1,2]. The O antigen is a region that provides a hydrophilic surface to the bacterial cell wall and is composed of lateral carbohydrate chains that vary in their composition This makes the O antigen useful in identifying bacteria [6,7]. Antibodies from the sera of immunized rabbits with SP287/3, SP459/1, SP308/3, and SP073/14 react against both their own peptide and the different LPSs. The Western blot test shows a sera reaction against both the lateral chains and the cores of the LPSs. The analysis of the human sera shows a response against the SPs and LPSs. Conclusion: The designed synthetic peptides are mimotopes of LPS epitopes of Salmonella and E. coli that possess immunogenic capacity. These mimotopes could be considered for use in the design of vaccines against both enterobacteria

Objectives

Methods

Results