title: The Escherichia coli hemolysin secretion apparatus--a versatile antigen delivery system in attenuated Salmonella.
abstract: The E. coli hemolysin (HlyA) secretion apparatus represents a type I secretion system that is fully functional in Salmonella. The system which consists of the two specific membrane proteins HlyB and HlyD and the outer membrane protein TolC, recognizes on HlyA a C-terminally located signal sequence of about 60 amino acids. Fusion proteins to which this signal sequence is covalently linked at the C-terminus are also recognized by this secretion apparatus. The efficiency of secretion is dependent on the rate of folding of the reporter protein. Secretion-competent regions of a given reporter protein that is not secretable as entire protein can be screened by a recently constructed transposon TnhlyAs which allows the insertion of the secretion signal into any region of the reporter protein. The genetic information for antigens of any source ranging in size between 10 and 1000 amino acids can be easily inserted into a recently constructed secretion vector which will allow the secretion of the fused antigen(s) in attenuated Salmonella typhimurium strains and in other attenuated Enterobacteriaceae. By manipulation of the Hly secretion system the antigen can be either completely secreted into the environment, fixed on the outer membrane or arrested in the cytoplasm of the used carrier strain. By the use of appropriate attenuated Salmonella strains the antigen is delivered in isolated compartments or to the cytosolic compartment. The extracellular delivery of such antigens is also possible with the help of appropriate carrier strains. The immunological consequences of the different display of the processed antigen will be discussed in the paper by Hess et al in this volume. With a similar antigen delivery system the easy identification and molecular characterization of unknown antigens recognized by the immune system in an infection is also feasible.
authors.affiliation: Theodor-Boveri-Institut für Biowissenschaften (Mikrobiologie), University of Würzburg, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Gentschev
authors.forename: G
authors.initials: G
authors.lastname: Dietrich
authors.forename: H J
authors.initials: HJ
authors.lastname: Mollenkopf
authors.forename: Z
authors.initials: Z
authors.lastname: Sokolovic
authors.forename: J
authors.initials: J
authors.lastname: Hess
authors.forename: S H
authors.initials: SH
authors.lastname: Kaufmann
authors.forename: W
authors.initials: W
authors.lastname: Goebel
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000276:immunology*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D001428:Bacterial Vaccines; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004755:Enterobacteriaceae / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D017930:Genes, Reporter; D005822:Genetic Vectors; D006460:Hemolysin Proteins / Q000096:biosynthesis*; D006801:Humans; D016366:Open Reading Frames; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D012475:Salmonella / Q000276:immunology*; D012486:Salmonella typhimurium / Q000276:immunology*; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D011993:Recombinant Fusion Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic