Chimeric Dr Fimbriae with a Herpes Simplex Virus Type 1 Epitope as a Model for a Recombinant Vaccine

Zalewska, Beata; Piątek, Rafał; Konopa, Graz ̇yna; Nowicki, B.; Nowicki, Stella; Kur, Józef

doi:10.1128/iai.71.10.5505-5513.2003

Cited by 16 publications

(30 citation statements)

References 24 publications

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“…For amplification of DNA fragment II, the following primers were used: 3-DraE-V5 forward primer GGCAAACCGATCCCGAAC-CCGCTGCTCGGCCTGGACTCCACCGCACCGCACCAATAGGGCC-GGTCACCGTGCAAGCG (the italicized sequence encodes a V5 epitope, and the bold part is complementary to the nucleotide sequence at the 39 end of the draE gene) and 4-DraE-HSV reverse primer (Zalewska et al, 2003). The DNA fragments obtained were hybridized and reamplified by PCR in a Perkin-Elmer 2400 thermocycler.…”

Section: Methodsmentioning

confidence: 99%

“…The draE gene with its signal sequence, encoding DraE adhesin, was amplified in two DNA fragments (I and II) on a template of pCC90, as previously described for insertion of an HSV-1 epitope (Zalewska et al, 2003). The primers used in the amplification were designed based on the sequencing data obtained (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid pDraE-HSV3, encoding DraE adhesin with an inserted glycoprotein D (gD) epitope (QPELAPEDPED) of herpes simplex virus type 1 (HSV-1), was constructed as described previously (Zalewska et al, 2003).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid pDraE-V5, encoding DraE adhesin with an inserted V5 epitope, was constructed on the basis of a pET30Ek/LIC expression vector as previously described for the HSV-1 gD epitope (Zalewska et al, 2003).…”

Section: Methodsmentioning

confidence: 99%

“…For amplification of DNA fragment I, the following primers were used: 1-DraE-HSV forward primer (Zalewska et al, 2003) and 2-DraE-V5 reverse primer GGTGGAGTCCAGGCCGAGCAGCGGGT-TCGGGATCGGTTTGCCGGTCAGGTCACCAACCCGTACCTGCAC-TC (the italicized sequence encodes a V5 epitope, and the bold part is complementary to the nucleotide sequence at the 59 end of the draE gene). For amplification of DNA fragment II, the following primers were used: 3-DraE-V5 forward primer GGCAAACCGATCCCGAAC-CCGCTGCTCGGCCTGGACTCCACCGCACCGCACCAATAGGGCC-GGTCACCGTGCAAGCG (the italicized sequence encodes a V5 epitope, and the bold part is complementary to the nucleotide sequence at the 39 end of the draE gene) and 4-DraE-HSV reverse primer (Zalewska et al, 2003).…”

Section: Methodsmentioning

confidence: 99%

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DAF- and collagen-binding properties of chimeric Dr fimbriae

et al. 2007

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Display of heterologous proteins on the surface of micro-organisms has become an increasingly used strategy for a range of applications in microbiology, biotechnology and vaccinology. In this study, the potential of the major structural protein DraE of Escherichia coli Dr fimbriae as a display system for heterologous sequences was tested. One copy of a heterologous sequence mimicking a small P k epitope of simian virus 5 was inserted into the draE gene, replacing the N-terminal region of the surface-exposed domain 2 as previously done with the glycoprotein D of herpes simplex virus type 1. The exposure of chimeric proteins on the bacterial surface was detected by immunofluorescence microscopy. Insertion of the heterogenic peptides had no detectable effect on the Ig-barrel structure of the DraE fimbrial subunits, as confirmed by FTIR spectroscopy. Additionally, the affinity of the chimeric fimbriae for DAF and type IV collagen was similar to that of the wild-type Dr fimbriae.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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DAF- and collagen-binding properties of chimeric Dr fimbriae

et al. 2007

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Disordered epitopes as peptide vaccines

et al. 2018

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The development of clinically useful peptide‐based vaccines remains a long‐standing goal. This review highlights that intrinsically disordered protein antigens, which lack an ordered three‐dimensional structure, represent excellent starting points for the development of such vaccines. Disordered proteins represent an important class of antigen in a wide range of human pathogens, and, contrary to widespread belief, they are frequently targets of protective antibody responses. Importantly, disordered epitopes appear invariably to be linear epitopes, rendering them ideally suited to incorporation into a peptide vaccine. Nonetheless, the conformational properties of disordered antigens, and hence their recognition by antibodies, frequently depend on the interactions they make and the context in which they are presented to the immune system. These effects must be considered in the design of an effective vaccine. Here we discuss these issues and propose design principles that may facilitate the development of peptide vaccines targeting disordered antigens.

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The DraC usher in Dr fimbriae biogenesis of uropathogenic E. coli Dr+ strains

et al. 2010

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Biogenesis of Dr fimbriae encoded by the dra gene cluster of uropathogenic Escherichia coli strains requires the chaperone-usher pathway. This secretion system is based on two non-structural assembly components, the DraB periplasmic chaperone and DraC outer-membrane usher. The DraB controls the folding of DraE subunits, and DraC forms the assembly and secretion platform for polymerization of subunits in linear fibers. In this study, mutagenesis of the DraC N-terminus was undertaken to select residues critical for Dr fimbriae bioassembly. The DraC-F4A, DraC-C64, DraC-C100A and DraC-W142A significantly reduced the adhesive ability of E. coli strains. The biological activity of the DraC mutants as a assembly platform for Dr fimbriae polymerization was verified by agglutination of human erythrocytes and adhesion to DAF localized at the surface of CHO-DAF(+) and HeLa cells. The residue F4 of the DraC usher conserved among FGL and FGS chaperone-assembled adhesive organelles can be used to design pillicides blocking the biogenesis of Dr fimbriae. Because the draC and afaC-III genes share 100% identity the range of the virulence determinant inhibitors could also be extended to E. coli strains encoding afa-3 gene cluster. The investigations performed showed that the usher N-terminus plays an important role in biogenesis of complete fiber.

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Chimeric Dr Fimbriae with a Herpes Simplex Virus Type 1 Epitope as a Model for a Recombinant Vaccine

Cited by 16 publications

References 24 publications

DAF- and collagen-binding properties of chimeric Dr fimbriae

DAF- and collagen-binding properties of chimeric Dr fimbriae

Disordered epitopes as peptide vaccines

The DraC usher in Dr fimbriae biogenesis of uropathogenic E. coli Dr+ strains

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