An autotransporter display platform for the development of multivalent recombinant bacterial vector vaccines.

Wouter Sp Jong,Jan-Willem De Gier,Corinne M Ten Hagen-Jongman,Guus F Rimmelzwaan,Maria H Daleke-Schermerhorn,Nicole N Van Der Wel,Karin De Punder,Frank Follmann,Carolien E Van De Sandt,Else Marie Agger,Joen Luirink,Peter Andersen,David Vikström

doi:10.1186/s12934-014-0162-8

Abstract

BackgroundThe Autotransporter pathway, ubiquitous in Gram-negative bacteria, allows the efficient secretion of large passenger proteins via a relatively simple mechanism. Capitalizing on its crystal structure, we have engineered the Escherichia coli autotransporter Hemoglobin protease (Hbp) into a versatile platform for secretion and surface display of multiple heterologous proteins in one carrier molecule.ResultsAs proof-of-concept, we demonstrate efficient secretion and high-density display of the sizeable Mycobacterium tuberculosis antigens ESAT6, Ag85B and Rv2660c in E. coli simultaneously. Furthermore, we show stable multivalent display of these antigens in an attenuated Salmonella Typhimurium strain upon chromosomal integration. To emphasize the versatility of the Hbp platform, we also demonstrate efficient expression of multiple sizeable antigenic fragments from Chlamydia trachomatis and the influenza A virus at the Salmonella cell surface.ConclusionsThe successful efficient cell surface display of multiple antigens from various pathogenic organisms highlights the potential of Hbp as a universal platform for the development of multivalent recombinant bacterial vector vaccines.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-014-0162-8) contains supplementary material, which is available to authorized users.

Highlights

Live attenuated strains of pathogenic bacteria that synthesize heterologous antigens are being developed as vaccines for several infectious diseases and cancer
ESAT6 folds into an α-helical hairpin [33], a relatively simple structure that was previously shown to be compatible with Hemoglobin protease (Hbp)-mediated translocation [9]
Cells were grown to early log-phase after which the expression of Hbp was induced by the addition of isopropyl β-Dthiogalactopyranoside (IPTG)

Summary

Introduction

Live attenuated strains of pathogenic bacteria that synthesize heterologous antigens are being developed as vaccines for several infectious diseases and cancer. Using Salmonella vaccine strains, cell surface display or secretion of heterologous antigens has been shown to yield superior immune responses compared to intracellular expression [2,3]. In Salmonella and other Gram-negative bacteria like Escherichia coli, efficient secretion and surface. The Autotransporter pathway [4,5], known as the Type Va secretion system [6], represents a ubiquitous and simple mechanism for protein translocation across the Gram-negative cell envelope and is typically used for the secretion of large virulence factors. The Autotransporter pathway, ubiquitous in Gram-negative bacteria, allows the efficient secretion of large passenger proteins via a relatively simple mechanism. Capitalizing on its crystal structure, we have engineered the Escherichia coli autotransporter Hemoglobin protease (Hbp) into a versatile platform for secretion and surface display of multiple heterologous proteins in one carrier molecule

Methods

Results

Discussion

Conclusion