Mucosal vaccination of mice with recombinant Proteus mirabilis structural fimbrial proteins

Scavone, Paola; Sosa, Vanessa; Pellegrino, Rafael; Galvalisi, Umberto; Zunino, Pablo

doi:10.1016/j.micinf.2004.04.006

Cited by 36 publications

(47 citation statements)

References 29 publications

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“…Whole-cell preparations obtained from 48 h static cultures in LB broth were run on SDS-PAGE and proteins were transferred to nitrocellulose membranes (Bio-Rad) as described by Towbin et al (1979). Western immunoblots were performed using a 1 : 200 dilution in PBS/Tween 20/1 % (w/v) skimmed milk of rabbit polyclonal immune sera raised against MrpA (Zunino et al, 2001), PmfA (Zunino et al, 2003) and AtfA (Zunino et al, 2000), and mouse polyclonal immune sera raised against UcaA (Scavone et al, 2004). Isogenic fimbrial mrpA, pmfA, atfA-C and ucaA mutants were included as controls in the Western blotting assays (strains MSD2, P2, A4 and UM1 respectively; see Table 1).…”

Section: Sds-pagementioning

confidence: 99%

Proteus mirabilis isolates of different origins do not show correlation with virulence attributes and can colonize the urinary tract of mice

2006

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Proteus mirabilis has been described as an aetiological agent in a wide range of infections, playing an important role in urinary tract infections (UTIs). In this study, a collection of P. mirabilis isolates obtained from clinical and non-clinical sources was analysed in order to determine a possible correlation between origin, virulence factors and in vivo infectivity. Isolates were characterized in vitro, assessing several virulence properties that had been previously associated with P. mirabilis uropathogenicity. Swarming motility, urease production, growth in urine, outer-membrane protein patterns, ability to grow in the presence of different iron sources, haemolysin and haemagglutinin production, and the presence and expression of diverse fimbrial genes, were analysed. In order to evaluate the infectivity of the different isolates, the experimental ascending UTI model in mice was used. Additionally, the Dienes test and the enterobacterial repetitive intergenic consensus (ERIC)-PCR assay were performed to assess the genetic diversity of the isolates. The results of the present study did not show any correlation between distribution of the diverse potential urovirulence factors and isolate source. No significant correlation was observed between infectivity and the origin of the isolates, since they all similarly colonized the urinary tract of the challenged mice. Finally, all isolates showed unique ERIC-PCR patterns, indicating that the isolates were genetically diverse. The results obtained in this study suggest that the source of P. mirabilis strains cannot be correlated with pathogenic attributes, and that the distribution of virulence factors between isolates of different origins may correspond to the opportunistic nature of the organism.

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Section: Sds-pagementioning

confidence: 99%

Proteus mirabilis isolates of different origins do not show correlation with virulence attributes and can colonize the urinary tract of mice

2006

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“…They aid in the process of infection by mediating adhesion, aggregation, hemagglutination, or motility in vivo (39). Their use in developing subunit vaccines against P. mirabilis has also been explored (45,46); however, phase variation of certain fimbriae requires that they be used only in conjunction with other proteins as part of a multivalent vaccine (26,59). Thus, identifying nonfimbrial surface proteins that contribute to the pathogen's persistence in vivo is critical.…”

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confidence: 99%

Adhesion, Invasion, and Agglutination Mediated by Two Trimeric Autotransporters in the Human Uropathogen Proteus mirabilis

et al. 2010

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Fimbriae of the human uropathogen Proteus mirabilis are the only characterized surface proteins that contribute to its virulence by mediating adhesion and invasion of the uroepithelia. PMI2122 (AipA) and PMI2575 (TaaP) are annotated in the genome of strain HI4320 as trimeric autotransporters with "adhesinlike" and "agglutinating adhesin-like" properties, respectively. The C-terminal 62 amino acids (aa) in AipA and 76 aa in TaaP are homologous to the translocator domains of YadA from Yersinia enterocolitica and Hia from Haemophilus influenzae. Comparative protein modeling using the Hia three-dimensional structure as a template predicted that each of these domains would contain four antiparallel beta sheets and that they formed homotrimers. Recombinant AipA and TaaP were seen as ϳ28 kDa and ϳ78 kDa, respectively, in Escherichia coli, and each also formed high-molecular-weight homotrimers, thus supporting this model. E. coli synthesizing AipA or TaaP bound to extracellular matrix proteins with a 10-to 60-fold-higher level of affinity than the control strain. Inactivation of aipA in P. mirabilis strains significantly (P < 0.01) reduced the mutants' ability to adhere to or invade HEK293 cell monolayers, and the functions were restored upon complementation. A 51-aa-long invasin region in the AipA passenger domain was required for this function. E. coli expressing TaaP mediated autoagglutination, and a taaP mutant of P. mirabilis showed significantly (P < 0.05) more reduced aggregation than HI4320. Gly-247 in AipA and Gly-708 in TaaP were indispensable for trimerization and activity. AipA and TaaP individually offered advantages to P. mirabilis in a murine model. This is the first report characterizing trimeric autotransporters in P. mirabilis as afimbrial surface adhesins and autoagglutinins.

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“…All bacterial suspensions were in PBS. Immunization was performed by instilling 10 ml bacterial suspension into each nostril (Pellegrino et al, 2003;Scavone et al, 2004Scavone et al, , 2009. Vaccination was performed twice on days 1 and 28 (Pickett et al, 2000).…”

Section: Methodsmentioning

confidence: 99%

Nasal immunization with attenuated Salmonella Typhimurium expressing an MrpA–TetC fusion protein significantly reduces Proteus mirabilis colonization in the mouse urinary tract

Scavone

Umpiérrez

Maskell

et al. 2011

Journal of Medical Microbiology

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Mucosal vaccination of mice with recombinant Proteus mirabilis structural fimbrial proteins

Cited by 36 publications

References 29 publications

Proteus mirabilis isolates of different origins do not show correlation with virulence attributes and can colonize the urinary tract of mice

Proteus mirabilis isolates of different origins do not show correlation with virulence attributes and can colonize the urinary tract of mice

Adhesion, Invasion, and Agglutination Mediated by Two Trimeric Autotransporters in the Human Uropathogen Proteus mirabilis

Nasal immunization with attenuated Salmonella Typhimurium expressing an MrpA–TetC fusion protein significantly reduces Proteus mirabilis colonization in the mouse urinary tract

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