Inhibition of
            <i>Salmonella enterica</i>
            Serovar Typhimurium Motility and Entry into Epithelial Cells by a Protective Antilipopolysaccharide Monoclonal Immunoglobulin A Antibody

Forbes, Stephen J.; Eschmann, Marisa; Mantis, Nicholas J.

doi:10.1128/iai.00416-08

Cited by 98 publications

(123 citation statements)

References 54 publications

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“…There is emerging evidence to suggest that protective anti-LPS IgA antibodies may have direct effects on microbial virulence, independent of agglutination (20)(21)(22)(23)(24). In the case of V. cholerae, Bishop and Camilli demonstrated using dark-field microscopy that polyclonal anti-LPS IgA antibodies inhibit V. cholerae motility in vitro, an observation that has been confirmed by others (13,14).…”

mentioning

confidence: 60%

“…The monoclonal dimeric IgA antibodies Sal4 (specific for the Salmonella enterica serovar Typhimurium O5 antigen) (29) and 2D6 (specific for V. cholerae OPS) (12) were obtained from Marian Neutra (Children's Hospital, Boston, MA). Sal4 and 2D6 hybridomas were maintained in CD hybridoma serum-free, protein-free medium (Gibco-Invitrogen, Carlsbad, CA) without antibiotics at 37°C in a 5% CO 2 -95% air atmosphere, as described previously (23). The concentrations of Sal4 and 2D6 were determined by sandwich enzyme-linked immunosorbent assay with IgA TEPC-15 (Sigma-Aldrich, St. Louis, MO) as a standard.…”

Section: Methodsmentioning

confidence: 99%

“…Visually, IgA-mediated motility arrest of V. cholerae occurred before the bacteria became agglutinated, suggesting that the two phenomena are distinct. In the case of Salmonella enterica serovar Typhimurium, we reported in 2008 that treatment of S. Typhimurium with a protective O-antigen-specific IgA monoclonal antibody (MAb) known as Sal4 results in the immediate arrest of flagellum-based motility, independent of agglutination (23). More recently we demonstrated that Sal4 triggers a reduction in type 3 secretion system (T3SS) activity and alterations in outer membrane integrity and onset of exopolysaccharide (EPS) production.…”

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confidence: 99%

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Rapid Effects of a Protective O-Polysaccharide-Specific Monoclonal IgA on Vibrio cholerae Agglutination, Motility, and Surface Morphology

2015

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b2D6 is a dimeric monoclonal immunoglobulin A (IgA) specific for the nonreducing terminal residue of Ogawa O-polysaccharide (OPS) of Vibrio cholerae. It was previously demonstrated that 2D6 IgA is sufficient to passively protect suckling mice from oral challenge with virulent V. cholerae O395. In this study, we sought to define the mechanism by which 2D6 IgA antibody protects the intestinal epithelium from V. cholerae infection. In a mouse ligated-ileal-loop assay, 2D6 IgA promoted V. cholerae agglutination in the intestinal lumen and limited the ability of the bacteria to associate with the epithelium, particularly within the crypt regions. In vitro fluorescence digital video microscopy analysis of antibody-treated V. cholerae in liquid medium revealed that 2D6 IgA not only induced the rapid (5-to 10-min) onset of agglutination but was an equally potent inhibitor of bacterial motility. Scanning electron microscopy showed that 2D6 IgA promoted flagellum-flagellum cross-linking, as well as flagellar entanglement with bacterial bodies, suggesting that motility arrest may be a consequence of flagellar tethering. However, monovalent 2D6 Fab fragments also inhibited V. cholerae motility, demonstrating that antibody-mediated agglutination and motility arrest are separate phenomena. While 2D6 IgA is neither bactericidal nor bacteriostatic, exposure of V. cholerae to 2D6 IgA (or Fab fragments) resulted in a 5-fold increase in surface-associated blebs, as well an onset of a wrinkled surface morphotype. We propose that the protective immunity conferred by 2D6 IgA is the result of multifactorial effects on V. cholerae, including agglutination, motility arrest, and possibly outer membrane stress. C holera is a life-threatening disease that remains endemic in many parts of the world (1-3). The etiological agent of cholera, Vibrio cholerae, is a noninvasive, Gram-negative bacterium that is acquired by humans via the fecal-oral route. Following ingestion, V. cholerae colonizes the mucosal surfaces of the small intestines, a process that is facilitated by the bacterium's single polar flagellum (4-7). Adherence to the epithelial surface requires expression of the toxin-coregulated pilus (TCP), in addition to other virulence factors (8), most notably, a potent ADP-ribosylating toxin known as cholera toxin (CT). CT disrupts chloride secretion within intestinal epithelial cells, inducing profuse water and electrolyte secretion and ultimately resulting in the hallmark "rice water diarrhea" associated with cholera. Cholera outbreaks frequently occur when water sanitation is disrupted, either following natural disasters or seasonally in areas where V. cholerae is endemic (9). The recent cholera outbreak in Haiti following the 2010 earthquake highlighted the ongoing potential of V. cholerae to cause mass causalities, as it resulted in more than half a million infected individuals and more than 7,000 deaths (10). Due to the rapid onset of symptoms and limited treatment options, control of cholera in many parts of the globe, particularly...

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confidence: 60%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Rapid Effects of a Protective O-Polysaccharide-Specific Monoclonal IgA on Vibrio cholerae Agglutination, Motility, and Surface Morphology

2015

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“…Additionally, the ⌬wbaP mutant (SKI12) showed a severe defect in motility. There are several other reports connecting LPS mutations with a loss in motility: in serovar Typhimurium, addition of a monoclonal anti-LPS antibody to a diluted bacterial culture led to instant paralysis of the bacteria (8), and in a Pseudomonas aeruginosa ⌬waaL strain, a severe impairment in motility was detected (2). Also, an Escherichia coli mutant lacking O antigen was reported to show a nonmotile phenotype (37).…”

Section: Discussionmentioning

confidence: 99%

O-Antigen-NegativeSalmonella entericaSerovar Typhimurium Is Attenuated in Intestinal Colonization but Elicits Colitis in Streptomycin-Treated Mice

et al. 2009

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Lipopolysaccharide (LPS) is a major constituent of the outer membrane and an important virulence factor of Salmonella enterica subspecies 1 serovar Typhimurium (serovar Typhimurium). To evaluate the role of LPS in eliciting intestinal inflammation in streptomycin-treated mice, we constructed an O-antigen-deficient serovar Typhimurium strain through deletion of the wbaP gene. The resulting strain was highly susceptible to human complement activity and the antimicrobial peptide mimic polymyxin B. Furthermore, it showed a severe defect in motility and an attenuated phenotype in a competitive mouse infection experiment, where the ⌬wbaP strain (SKI12) was directly compared to wild-type Salmonella. Nevertheless, the ⌬wbaP strain (SKI12) efficiently invaded HeLa cells in vitro and elicited acute intestinal inflammation in streptomycin-pretreated mice. Our experiments prove that the presence of complete LPS is not essential for in vitro invasion or for triggering acute colitis.

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“…Swimming assays were performed on 0.3% agar plates as described in ref. 60 with slight modifications; see SI Appendix, Materials and Methods for details. Swarming assays were performed on 0.5% agar plates.…”

Section: Methodsmentioning

confidence: 99%

RNA target profiles direct the discovery of virulence functions for the cold-shock proteins CspC and CspE

Michaux

Holmqvist

Vasicek

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

115

152

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The functions of many bacterial RNA-binding proteins remain obscure because of a lack of knowledge of their cellular ligands. Although well-studied cold-shock protein A (CspA) family members are induced and function at low temperature, others are highly expressed in infection-relevant conditions. Here, we have profiled transcripts bound in vivo by the CspA family members of Salmonella enterica serovar Typhimurium to link the constitutively expressed CspC and CspE proteins with virulence pathways. Phenotypic assays in vitro demonstrated a crucial role for these proteins in membrane stress, motility, and biofilm formation. Moreover, double deletion of cspC and cspE fully attenuates Salmonella in systemic mouse infection. In other words, the RNA ligand-centric approach taken here overcomes a problematic molecular redundancy of CspC and CspE that likely explains why these proteins have evaded selection in previous virulence factor screens in animals. Our results highlight RNA-binding proteins as regulators of pathogenicity and potential targets of antimicrobial therapy. They also suggest that globally acting RNA-binding proteins are more common in bacteria than currently appreciated.RNA-binding protein | cold-shock protein | Salmonella | bacterial pathogenesis | stress response T he myriad of coding and noncoding RNAs in a cell generally do not act in isolation but rapidly associate with RNA-binding proteins (RBPs) to execute their functions. Recent methods relying on the global capture of polyadenylated transcripts in eukaryotes have dramatically expanded our knowledge of RBP activity. These approaches have revealed many previously unsuspected RBPs, suggesting a much wider scope of posttranscriptional control based on thousands of new RBP-mRNA interactions (1, 2). Nevertheless, the functions of many of these newly discovered RBPs remain unclear.In contrast to the situation in eukaryotes, there is a paucity of knowledge about RBPs in prokaryotes. This lack of knowledge is compounded further by the lack of a poly(A) tail on functional transcripts, which precludes similar global discovery studies of bacterial RBP networks. Extensive profiling of cellular targets of the small RNA (sRNA) chaperone Hfq and the translational repressor CsrA (3-7) have shown that large posttranscriptional networks also exist in bacteria. In addition, the ProQ protein has recently been identified as a previously overlooked global RBP in Salmonella enterica (8). As new methods are developed to identify RPBs in bacteria globally, it is essential to be able to analyze their functions systematically to understand their cellular and physiological roles (9).Cold-shock proteins (CSPs) constitute the largest nonribosomal RBP family in Gram-negative bacteria, including the model species Escherichia coli and Salmonella enterica. Their conserved nucleic acid-binding cold-shock domain makes them members of a widespread RBP superfamily that includes the well-investigated eukaryotic Y-box proteins (10, 11). Of the nine and six CSP paralogs present in E. c...

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Inhibition of Salmonella enterica Serovar Typhimurium Motility and Entry into Epithelial Cells by a Protective Antilipopolysaccharide Monoclonal Immunoglobulin A Antibody

Cited by 98 publications

References 54 publications

Rapid Effects of a Protective O-Polysaccharide-Specific Monoclonal IgA on Vibrio cholerae Agglutination, Motility, and Surface Morphology

Rapid Effects of a Protective O-Polysaccharide-Specific Monoclonal IgA on Vibrio cholerae Agglutination, Motility, and Surface Morphology

O-Antigen-NegativeSalmonella entericaSerovar Typhimurium Is Attenuated in Intestinal Colonization but Elicits Colitis in Streptomycin-Treated Mice

RNA target profiles direct the discovery of virulence functions for the cold-shock proteins CspC and CspE

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