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Attachment of Yersinia pestis to human respiratory cell lines is inhibited by certain oligosaccharidesPneumonic plague is an aggressive disease that is clinically difficult to treat. Inhibition of attachment using oligosaccharide receptor mimics may provide an alternative to antibiotics. The virulent Yersinia pestis strain GB was demonstrated to attach to the murine monocyte cell line (J774A.1) and a range of human respiratory epithelial cell lines: nasal (RPMI-2650), bronchial (BEAS2-B) and alveolar (A549). Attachment was greatest to the A549 and BEAS2-B cell lines. Pre-treatment of the cell lines with tunicamycin reduced attachment by 55-65 %, indicating the importance of cell-surface carbohydrates in adhesion. The cell lines displayed differences in the oligosaccharides that inhibited attachment. p-Nitrophenol was the best inhibitor for each cell line. Disaccharides such as GalNAcb1-3Gal and GalNAcb1-4Gal were also good inhibitors, particularly for the RPMI-2650 cell line. This demonstrates the potential of oligosaccharides as potential anti-adhesion therapeutics. INTRODUCTIONYersinia pestis is the aetiological agent of 'black death', a pandemic that swept across Asia and Europe between the 14th and 17th centuries causing millions of deaths (Perry & Fetherston, 1997 Russell et al., 1998;Steward et al., 2004). However, recently there has been a worrying increase in multidrug-resistant strains (Galimand et al., 1997;Wong et al., 2000).The infectious nature and high mortality rate of pneumonic plague means that, even with today's medical advances, it remains a disease to be feared. High aerosol transmissibility makes plague a high-priority agent for potential bioterrorists (Inglesby et al., 2000). A novel preventative mechanism of plague infection may be via inhibiting adhesion of Y. pestis to the respiratory tract. Bacteria generally attach to mammalian cells using proteins, glycoproteins and glycolipids located on the mammalian cell membrane. In Y. pestis, identified putative adhesins include the pH6 antigen, plasminogen activator, LPS and the capsular F1 antigen (Lindler et al., 1990;Kienle et al., 1992;Lindler & Tall, 1993;Straley, 1993; Lähteenmaki et al., 1998;Payne et al., 1998). The potential of short-chain oligosaccharide receptor mimics to inhibit attachment has been investigated with varying success in many bacterial species, both in vitro (Krivan et al., 1988;Hambrook et al., 2004; Thomas & Brooks, 2004a, b) and in vivo (Idänpään-Heikkilä et al., 1997;Bryan et al., 1999). This study investigated the attachment of Y. pestis strain GB to a range of human respiratory epithelial cell lines. The ability of a range of oligosaccharides to prevent attachment was determined. METHODSMaterials. Tissue culture reagents, p-nitrophenol and tunicamycin were purchased from Sigma-Aldrich. Saccharides were purchased as powders from Dextra Laboratories or Sigma-Aldrich. Anti-rabbit IgG-FITC conjugate was purchased from Sigma-Aldrich. Rabbit anti-ganglioside antibodies were purchased from Calbiochem.Abbreviations: P...
Attachment of Yersinia pestis to human respiratory cell lines is inhibited by certain oligosaccharidesPneumonic plague is an aggressive disease that is clinically difficult to treat. Inhibition of attachment using oligosaccharide receptor mimics may provide an alternative to antibiotics. The virulent Yersinia pestis strain GB was demonstrated to attach to the murine monocyte cell line (J774A.1) and a range of human respiratory epithelial cell lines: nasal (RPMI-2650), bronchial (BEAS2-B) and alveolar (A549). Attachment was greatest to the A549 and BEAS2-B cell lines. Pre-treatment of the cell lines with tunicamycin reduced attachment by 55-65 %, indicating the importance of cell-surface carbohydrates in adhesion. The cell lines displayed differences in the oligosaccharides that inhibited attachment. p-Nitrophenol was the best inhibitor for each cell line. Disaccharides such as GalNAcb1-3Gal and GalNAcb1-4Gal were also good inhibitors, particularly for the RPMI-2650 cell line. This demonstrates the potential of oligosaccharides as potential anti-adhesion therapeutics. INTRODUCTIONYersinia pestis is the aetiological agent of 'black death', a pandemic that swept across Asia and Europe between the 14th and 17th centuries causing millions of deaths (Perry & Fetherston, 1997 Russell et al., 1998;Steward et al., 2004). However, recently there has been a worrying increase in multidrug-resistant strains (Galimand et al., 1997;Wong et al., 2000).The infectious nature and high mortality rate of pneumonic plague means that, even with today's medical advances, it remains a disease to be feared. High aerosol transmissibility makes plague a high-priority agent for potential bioterrorists (Inglesby et al., 2000). A novel preventative mechanism of plague infection may be via inhibiting adhesion of Y. pestis to the respiratory tract. Bacteria generally attach to mammalian cells using proteins, glycoproteins and glycolipids located on the mammalian cell membrane. In Y. pestis, identified putative adhesins include the pH6 antigen, plasminogen activator, LPS and the capsular F1 antigen (Lindler et al., 1990;Kienle et al., 1992;Lindler & Tall, 1993;Straley, 1993; Lähteenmaki et al., 1998;Payne et al., 1998). The potential of short-chain oligosaccharide receptor mimics to inhibit attachment has been investigated with varying success in many bacterial species, both in vitro (Krivan et al., 1988;Hambrook et al., 2004; Thomas & Brooks, 2004a, b) and in vivo (Idänpään-Heikkilä et al., 1997;Bryan et al., 1999). This study investigated the attachment of Y. pestis strain GB to a range of human respiratory epithelial cell lines. The ability of a range of oligosaccharides to prevent attachment was determined. METHODSMaterials. Tissue culture reagents, p-nitrophenol and tunicamycin were purchased from Sigma-Aldrich. Saccharides were purchased as powders from Dextra Laboratories or Sigma-Aldrich. Anti-rabbit IgG-FITC conjugate was purchased from Sigma-Aldrich. Rabbit anti-ganglioside antibodies were purchased from Calbiochem.Abbreviations: P...
Seventy-two barrows (Landrace × Large White, initial BW of 4.9 ± 0.3 kg and 17 ± 3 d old) were used to determine if dietary chito-oligosaccharides can replace antibiotics as a means to reduce signs associated with infection in weaned pigs challenged with Escherichia coli. Pigs were assigned to 1 of 4 treatments in a randomized complete block design using 6 pens per treatment with 3 pigs per pen. The treatments consisted of pigs fed the unsupplemented corn-soybean meal diet challenged or unchallenged with E. coli K88 and pigs fed the same diet supplemented with 160 mg of chito-oligosaccharides or 100 mg of cyadox/kg and challenged with E. coli K88. On d 7, 1 group of pigs fed the unsupplemented diet, as well as all pigs fed diets containing chito-oligosaccharides or cyadox, were orally dosed with 30 mL of an alkaline broth containing E. coli K88. Another group of pigs fed the unsupplemented diet was orally dosed with 30 mL of sterilized alkaline broth. Fecal consistency was visually assessed each morning from d 7 to 14. Blood samples were collected at 0, 24, 48, and 168 h postinfection. On d 14 postchallenge, all pigs were killed to evaluate intestinal morphology and determine E. coli concentrations in the intestine. During the postchallenge period (wk 2), unsupplemented pigs challenged with E. coli had decreased (P < 0.05) BW gain, feed intake, fecal consistency, villus height, villus height:crypt depth ratio, and plasma IGF-1, and increased (P < 0.05) diarrhea incidence, E. coli counts in the intestine, plasma interleukin-1β, plasma IL-10, and IGA-positive cells in the jejunal and ileal lamina propria, compared with unchallenged pigs. Supplementation with cyadox largely mitigated these effects. Although chito-oligosaccharide reduced the incidence of diarrhea, the growth performance of E. coli-challenged pigs supplemented with chito-oligosaccharide was not better than that of unsupplemented pigs challenged with E. coli. Therefore, chito-oligosaccharide, at the amount used in this experiment, does not seem to be an effective substitute for antibiotics as a growth promoter for newly weaned pigs challenged with E. coli.
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