157 research outputs found
Duality of β-glucan microparticles: antigen carrier and immunostimulants
Designing efficient recombinant mucosal vaccines against enteric diseases is still a major challenge. Mucosal delivery of recombinant vaccines requires encapsulation in potent immunostimulatory particles to induce an efficient immune response. This paper evaluates the capacity of beta-glucan microparticles (GPs) as antigen vehicles and characterizes their immune-stimulatory effects. The relevant infectious antigen FedF was chosen to be loaded inside the microparticles. The incorporation of FedF inside the particles was highly efficient (roughly 85%) and occurred without antigen degradation. In addition, these GPs have immunostimulatory effects as well, demonstrated by the strong reactive oxygen species (ROS) production by porcine neutrophils upon their recognition. Although antigen-loaded GPs still induce ROS production, antigen loading decreases this production by neutrophils for reasons yet unknown. However, these antigen-loaded GPs are still able to bind their specific beta-glucan receptor, demonstrated by blocking complement receptor 3, which is the major beta-glucan receptor on porcine neutrophils. The dual character of these particles is confirmed by a T-cell proliferation assay. FedF-loaded particles induce a significantly higher FedF-specific T-cell proliferation than soluble FedF. Taken together, these results show that GPs are efficient antigen carriers with immune-stimulatory properties
An Efficient Algorithm For Simulating Fracture Using Large Fuse Networks
The high computational cost involved in modeling of the progressive fracture
simulations using large discrete lattice networks stems from the requirement to
solve {\it a new large set of linear equations} every time a new lattice bond
is broken. To address this problem, we propose an algorithm that combines the
multiple-rank sparse Cholesky downdating algorithm with the rank-p inverse
updating algorithm based on the Sherman-Morrison-Woodbury formula for the
simulation of progressive fracture in disordered quasi-brittle materials using
discrete lattice networks. Using the present algorithm, the computational
complexity of solving the new set of linear equations after breaking a bond
reduces to the same order as that of a simple {\it backsolve} (forward
elimination and backward substitution) {\it using the already LU factored
matrix}. That is, the computational cost is , where denotes the number of non-zeros of the Cholesky factorization of
the stiffness matrix . This algorithm using the direct sparse solver
is faster than the Fourier accelerated preconditioned conjugate gradient (PCG)
iterative solvers, and eliminates the {\it critical slowing down} associated
with the iterative solvers that is especially severe close to the critical
points. Numerical results using random resistor networks substantiate the
efficiency of the present algorithm.Comment: 15 pages including 1 figure. On page pp11407 of the original paper
(J. Phys. A: Math. Gen. 36 (2003) 11403-11412), Eqs. 11 and 12 were
misprinted that went unnoticed during the proof reading stag
Seed produced anti ETEC antibodies for oral passive immunisation of piglets against post weaning diarrhoea
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Nanobody mediated inhibition of attachment of F18 fimbriae expressing Escherichia coli
Post-weaning diarrhea and edema disease caused by F18 fimbriated E. coli are important diseases in newly weaned piglets and lead to severe production losses in farming industry. Protective treatments against these infections have thus far limited efficacy. In this study we generated nanobodies directed against the lectin domain of the F18 fimbrial adhesin FedF and showed in an in vitro adherence assay that four unique nanobodies inhibit the attachment of F18 fimbriated E. coli bacteria to piglet enterocytes. Crystallization of the FedF lectin domain with the most potent inhibitory nanobodies revealed their mechanism of action. These either competed with the binding of the blood group antigen receptor on the FedF surface or induced a conformational change in which the CDR3 region of the nanobody displaces the D ''-E loop adjacent to the binding site. This D ''-E loop was previously shown to be required for the interaction between F18 fimbriated bacteria and blood group antigen receptors in a membrane context. This work demonstrates the feasibility of inhibiting the attachment of fimbriated pathogens by employing nanobodies directed against the adhesin domain
Simplified monomeric VHH-Fc antibodies provide new opportunities for passive immunization
Simplified monomeric monoclonal antibodies consisting of a single-domain VHH, derived from camelid heavy-chain only antibodies, fused with the Fc domain of either IgG (VHH-IgG) or IgA (VHH-IgA) antibodies, are promising therapeutic proteins. These simplified single-gene encoded antibodies are much easier to manufacture and can be produced in plants and in yeast for bulk applications. These merits enable novel passive immunization applications, such as in-feed oral delivery of VHH-IgAs, which have successfully provided protection against a gastrointestinal infection in the piglet model
O157:H7 and O104:H4 Vero/Shiga toxin-producing Escherichia coli outbreaks: respective role of cattle and humans
An enteroaggregative Verotoxin (Vtx)-producing Escherichia coli strain of serotype O104:H4 has recently been associated with an outbreak of haemolytic-uremic syndrome and bloody diarrhoea in humans mainly in Germany, but also in 14 other European countries, USA and Canada. This O104:H4 E. coli strain has often been described as an enterohaemorrhagic E. coli (EHEC), i.e. a Vtx-producing E. coli with attaching and effacing properties. Although both EHEC and the German O104:H4 E. coli strains indeed produce Vtx, they nevertheless differ in several other virulence traits, as well as in epidemiological characteristics. For instance, the primary sources and vehicles of typical EHEC infections in humans are ruminants, whereas no animal reservoir has been identified for enteroaggregative E. coli (EAggEC). The present article is introduced by a brief overview of the main characteristics of Vtx-producing E. coli and EAggEC. Thereafter, the O104:H4 E. coli outbreak is compared to typical EHEC outbreaks and the virulence factors and host specificity of EHEC and EAggEC are discussed. Finally, a renewed nomenclature of Vtx-producing E. coli is proposed to avoid more confusion in communication during future outbreaks and to replace the acronym EHEC that only refers to a clinical condition
Reproducible gene targeting in recalcitrant Escherichia coli isolates
<p>Abstract</p> <p>Background</p> <p>A number of allele replacement methods can be used to mutate bacterial genes. For instance, the Red recombinase system of phage Lambda has been used very efficiently to inactivate chromosomal genes in <it>E. coli </it>K-12, through recombination between regions of homology. However, this method does not work reproducibly in some clinical <it>E. coli </it>isolates.</p> <p>Findings</p> <p>The procedure was modified by using longer homologous regions (85 bp and 500-600 bp), to inactivate genes in the uropathogenic <it>E. coli </it>strain UTI89. An <it>lrhA </it>regulator mutant, and deletions of the <it>lac </it>operon as well as the complete <it>type 1 </it>fimbrial gene cluster, were obtained reproducibly. The modified method is also functional in other recalcitrant <it>E. coli</it>, like the avian pathogenic <it>E. coli </it>strain APEC1. The <it>lrhA </it>regulator and <it>lac </it>operon deletion mutants of APEC1 were successfully constructed in the same way as the UTI89 mutants. In other avian pathogenic <it>E. coli </it>strains (APEC3E, APEC11A and APEC16A) it was very difficult or impossible to construct these mutants, with the original Red recombinase-based method, with a Red recombinase-based method using longer (85 bp) homologous regions or with our modified protocol, using 500 - 600 bp homologous regions.</p> <p>Conclusions</p> <p>The method using 500-600 bp homologous regions can be used reliably in some clinical isolates, to delete single genes or entire operons by homologous recombination. However, it does not invariably show a greater efficiency in obtaining mutants, when compared to the original Red-mediated gene targeting method or to the gene targeting method with 85 bp homologous regions. Therefore the length of the homology regions is not the only limiting factor for the construction of mutants in these recalcitrant strains.</p
Plasmid parB contributes to uropathogenic Escherichia coli colonization in vivo by acting on biofilm formation and global gene regulation
The endogenous plasmid pUTI89 harbored by the uropathogenic Escherichia coli (UPEC) strain UTI89 plays an important role in the acute stage of infection. The partitioning gene parB is important for stable inheritance of pUTI89. However, the function of partitioning genes located on the plasmid in pathogenesis of UPEC still needs to be further investigated. In the present study, we observed that disruption of the parB gene leads to a deficiency in biofilm formation in vitro. Moreover, in a mixed infection with the wild type strain and the parB mutant, in an ascending UTI mouse model, the mutant displayed a lower bacterial burden in the bladder and kidneys, not only at the acute infection stage but also extending to 72 hours post infection. However, in the single infection test, the reduced colonization ability of the parB mutant was only observed at six hpi in the bladder, but not in the kidneys. The colonization capacity in vivo of the parB-complemented strain was recovered. qRT-PCR assay suggested that ParB could be a global regulator, influencing the expression of genes located on both the endogenous plasmid and chromosome, while the gene parA or the operon parAB could not. Our study demonstrates that parB contributes to the virulence of UPEC by influencing biofilm formation and proposes that the parB gene of the endogenous plasmid could regulate gene expression globally
Passive mucosal immunisation with novel simplified IgA antibodies produced in seeds prevents enterotoxigenic Escherichia coli (ETEC) infection in weaned piglets
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Structural insight in the inhibition of adherence of F4 fimbriae producing enterotoxigenic Escherichia coli by llama single domain antibodies
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