crip.umontreal.ca - Université de Montréal

Recommendations

Info

Présentation orale no. 2 Page 21 Implication du LOS d’Haemophilus parasuis lors des interactions avec des cellules endothéliales porcines de microvaisseaux cérébraux B. Bouchet, , G. Vanier, M. Jacques, M. Gottschalk Groupe de Recherche sur les maladies infectieuses du Porc (GREMIP) Faculté de médecine vétérinaire, Saint-Hyacinthe, QC Haemophilus parasuis est un pathogène du porc causant la maladie de Glässer chez les porcelets. Les animaux infectés présentent des signes cliniques de polysérosite fibrineuse, de polyarthrite et de méningite. À ce jour, peu de facteurs de virulence ont été décrits. De plus, la pathogenèse de l’infection causée par ce pathogène est encore mal connue. Pour accéder au système nerveux central et causer la méningite, H. parasuis pourrait traverser la barrière hémato-méningée (BHM). En effet, H. parasuis est capable d’adhérer à et d’envahir des cellules endothéliales porcines de microvaisseaux cérébraux (PBMEC) provenant de la BHM. Le but de cette étude était (a) d’évaluer le rôle du lipooligosaccharide (LOS) de H. parasuis dans l’adhésion aux PBMEC, (b) d’étudier la capacité de H. parasuis à activer les PBMEC, et (c) d’évaluer le rôle du LOS dans cette activation. Les résultats ont démontré que l’adhésion de H. parasuis aux PBMEC était en partie médiée par le LOS. En effet, le LOS purifié réduit significativement l’adhésion du pathogène aux cellules. De plus, nous avons démontré qu’H. parasuis était capable d’activer les cellules endothéliales, cette activation se traduisant par la sécrétion d’interleukine-6 et d’interleukine-8. Le LOS de H. parasuis était responsable d’une faible partie de cette activation. Différentes souches de champ d’H. parasuis de sérotypes 4 et 5 (les sérotypes les plus prévalents en Amérique du Nord) ont stimulé les PBMEC à produire ces médiateurs de l’inflammation à des niveaux similaires. En général, les résultats suggèrent que le LOS de H. parasuis pourrait jouer un certain rôle dans la pathogenèse de la méningite causée par ce pathogène, tandis que d’autres composantes bactériennes pourraient également être impliquées dans ce processus.
Page 22 Présentation orale no. 3 Hemoglobin binding protein from Actinobacillus pleuropneumoniae: a novel method for extraction and isolation , Mario Jacques ** , and James W. Coulton * Dora Maria Skaf * , Mario Jacques ** , and James W. Coulton * *Department of Microbiology and Immunology, McGill University, 3775 University Street, Montreal, QC, Canada H3A 2B4 ** Groupe de recherche sur les maladies infectieuses du porc, Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, Canada J2S 7C6. Disease accounts for a multimillion dollar annual loss to the Canadian pork industry. Actinobacillus pleuropneumoniae (APP) is the etiological agent of a highly contagious and devastating porcine disease designated pleuropneumonia. Currently there is no vaccine that is consistently cross protective against all disease-associated strains of this bacterium. An attractive candidate for cross protective vaccine design is the conserved 105 kDa outer membrane hemoglobin binding protein (HgbA). HgbA is expressed by all serotypes of APP, allowing it to overcome bacteriostatic and bacteriocidal effects of the iron restricted environment in the host and thus contributing to APP pathogenicity. Deletion of hgbA results in disease attenuation. The requirement of HgbA for virulence and its exposure to the immune system at the bacterial cell surface make HgbA an ideal vaccine target. Our objective was to produce significant quantities of HgbA, purified to homogeneity. Recombinant HgbA with an appended C-terminal 6xHis tag was over-expressed by IPTG induction of the pET vector pRSC05 harboured by Escherichia coli BL21(DE3). Following cell lysis, total membranes were isolated by high speed ultracentrifugation (100 000 x g), and solubilized with 1% Sarkosyl, 150 mM NaCl, 50 mM Tris pH 7.5 at 4 oC. Sarkosyl-insoluble fractions were obtained with high speed ultracentrifugation and re-solubilzed in the 1% Sarkosyl solution at 70 o Disease accounts for a multimillion dollar annual loss to the Canadian pork industry. Actinobacillus pleuropneumoniae (APP) is the etiological agent of a highly contagious and devastating porcine disease designated pleuropneumonia. Currently there is no vaccine that is consistently cross protective against all disease-associated strains of this bacterium. An attractive candidate for cross protective vaccine design is the conserved 105 kDa outer membrane hemoglobin binding protein (HgbA). HgbA is expressed by all serotypes of APP, allowing it to overcome bacteriostatic and bacteriocidal effects of the iron restricted environment in the host and thus contributing to APP pathogenicity. Deletion of hgbA results in disease attenuation. The requirement of HgbA for virulence and its exposure to the immune system at the bacterial cell surface make HgbA an ideal vaccine target. Our objective was to produce significant quantities of HgbA, purified to homogeneity. Recombinant HgbA with an appended C-terminal 6xHis tag was over-expressed by IPTG induction of the pET vector pRSC05 harboured by Escherichia coli BL21(DE3). Following cell lysis, total membranes were isolated by high speed ultracentrifugation (100 000 x g), and solubilized with 1% Sarkosyl, 150 mM NaCl, 50 mM Tris pH 7.5 at 4 C. Following a third round of ultracentrifugation, HgbA was markedly enriched in the soluble fraction. Structural integrity of isolated HgbA may be assessed by its specificity for hemoglobin using a hemoglobin-agarose column. This extraction protocol produces 5 mg HgbA/ litre of culture at a quality that is near homogeneity even prior to column purification. Such high quality preparations of HgbA are advantageous for vaccine studies as well as for structural studies. oC. Sarkosyl-insoluble fractions were obtained with high speed ultracentrifugation and re-solubilzed in the 1% Sarkosyl solution at 70 oC. Following a third round of ultracentrifugation, HgbA was markedly enriched in the soluble fraction. Structural integrity of isolated HgbA may be assessed by its specificity for hemoglobin using a hemoglobin-agarose column. This extraction protocol produces 5 mg HgbA/ litre of culture at a quality that is near homogeneity even prior to column purification. Such high quality preparations of HgbA are advantageous for vaccine studies as well as for structural studies.
Page 1 and 2: 1 er Symposium du Centre de recherc
Page 3 and 4: Table des matières 1. Programme ..
Page 5 and 6: Session 2: Streptococcus suis Table
Page 7 and 8: Page 8 10 h 45 — 11 h 5-Truncatio
Page 9 and 10: Page 10 15 h — 15 h 15 14-Déterm
Page 11 and 12: Page 12 9 h 15 — 9 h 30 21- Compa
Page 13 and 14: Page 14 14 h 45 — 15 h 30- Étude
Page 15 and 16: Page 16 SESSION 1 : FAMILLE DES PAS
Page 17 and 18: Page 18 12-Intérêt des polymères
Page 19: Page 20 Présentation orale no. 1
Page 23 and 24: Page 24 Présentation orale no. 5 T
Page 25 and 26: Page 26 Présentation orale no. 7
Page 33 and 34: Page 34 Résumés/Abstracts Delegat
Page 37 and 38: Page 38
Page 43 and 44: Page 44 Résumés/Abstracts Prés e
Page 49 and 50: Résumés/Abstracts Delegates/Parti
Page 51 and 52: Présentation orale no. 29 Résumé
Page 59 and 60: Page 60 Titre princi pal Affiche no
Page 61 and 62: Affiche no. 4 Titre Remarque: Titre
Page 63 and 64: Page 64 Affiche no. 6 Propriétés
Page 65 and 66: Page 66 Affiche no. 8 Formulation d
Page 67 and 68: Page 68 Affiche no. 10 Carboxyméth
Page 69 and 70: Page 70 Affiche no. 12 Intérêt de
Page 71 and 72:
Page 72 Affiche no. 14 Pharmacocin
Page 73 and 74:
Page 74 Affiche no. 16 Développeme
Page 75 and 76:
Page 76 Affiche no. 18 Global Gene
Page 77 and 78:
Page 78 Affiche no. 20 A surface pl
Page 79 and 80:
Nous remercions pour leur contribut
show all

crip.umontreal.ca - Université de Montréal

Create successful ePaper yourself

Delete template?

Save as template?