Proceedings of the 10th International Colloquium on Paratuberculosis

More documents

Recommendations

Info

#134 Analysis <strong>of</strong> <strong>the</strong> vaccine potential <strong>of</strong> nine MAP specific proteins, identified by immunoproteomics and in silico bio-informatic screening, in a murine model Virginie Roupie, Sophie Viart, Ruddy Wattiez, Jean-Jacques Letesson, Kris Huygen WIV-ex Pasteur Institute Brussels, Belgium; University <strong>of</strong> Mons-Hainaut, Belgium; Fac.Univ. Notre-Dame de la Paîx Namur, Belgium Objective: Control <strong>of</strong> Mycobacterium avium subsp. paratuberculosis (MAP) is seriously hampered by <strong>the</strong> lack <strong>of</strong> adequate diagnostic tools, vaccines and <strong>the</strong>rapies. In this study, we have evaluated <strong>the</strong> vaccine potential <strong>of</strong> nine MAP proteins: MAP3199, MAP2677c, MAP1693c, previously identified by immunoproteomic analysis <strong>of</strong> MAP secretome [1] and Ag5, Ag6 (not-annotated), MAP1637c, MAP0388, MAP3743 and MAP3744 identified by bioinformatic in silico screening <strong>of</strong> <strong>the</strong> MAP genome. Materials and Methods: BALB/c and C57BL/6 mice were vaccinated with plasmid DNA encoding <strong>the</strong> nine selected candidates as described before [2]. Three weeks after <strong>the</strong> last immunisation, Th1 type cytokine responses (IL-2 and IFN-γ) against <strong>the</strong> respective purified recombinant proteins were tested on spleens from individual mice. Levels <strong>of</strong> antigen-specific total immunoglobulin G (IgG), IgG1, IgG2a and IgG2b antibodies were determined by ELISA. Six weeks after <strong>the</strong> last immunization, mice were challenged intravenously with luminescent MAP ATCC 19698 (2X106 CFU/mice). Mice were sacrificed and <strong>the</strong> number <strong>of</strong> bioluminescent bacteria was determined in spleen and liver homogenates at 4, 8 and 12 weeks after challenge [3]. Results: Vaccination with DNA encoding MAP1637c (predicted carboxylase) induced <strong>the</strong> strongest Th1 type immune responses, both in BALB/c and C57BL/6 mice, confirming previous findings on <strong>the</strong> potential <strong>of</strong> this in silico selected antigen for <strong>the</strong> serodiagnosis <strong>of</strong> bovine paratuberculosis in cattle [4]. Protective potential <strong>of</strong> <strong>the</strong> nine antigens was weaker than <strong>the</strong> one we have reported for <strong>the</strong> putative transglycosylase MAP0586c [2]. Conclusion: Plasmid vaccination, coupled to <strong>the</strong> use <strong>of</strong> bioluminescent MAP is a powerful tool for <strong>the</strong> screening <strong>of</strong> <strong>the</strong> vaccine potential <strong>of</strong> paratuberculosis antigens in mice. References: 1. Leroy, B., et al., Proteomics, 2007. 7(7): p. 1164-76. 2. Roupie, V., et al., Vaccine, 2008. 26: p. 4783-4794. 3. Rosseels, V., et al., Infect Immun, 2006. 74(6): p. 3684-6. 4. Leroy, B., et al., Vet. Microbiol., 2009 135(3-4): p. 313-319. 133
#153 SNP discovery in bovine Sp110 gene and its genetic association with infetion by MAP Otsanda Ruiz, Mikel Iriondo, Carmen Manzano, Iratxe Montes, Elena Molina, Joseba M. Garrido, Ramon A. Juste, Patricia Vazquez, Ad Koets, Andone Estonba University <strong>of</strong> <strong>the</strong> Basque Country, Spain; NEIKER-Tecnalia, Spain; Utrecht University, The Ne<strong>the</strong>rlands ; Three polymorphisms <strong>of</strong> <strong>the</strong> Sp110 gene have been associated with tuberculosis susceptibility in humans (Tosh et al. 2006). Moreover, Ipr1 gene in mice, a Sp110 homolog, has been reported to confer genetic resistance to Mycobacterium tuberculosis and Listeria monocytogenes through apoptotic pathway activation in infected macrophages (Pan et al. 2005). Therefore, we hypo<strong>the</strong>size that <strong>the</strong> bovine Sp110 gene could be involved in <strong>the</strong> innate immunity response against Mycobacterium avium subsp. paratuberculosis (MAP) infection in cattle. To test <strong>the</strong> hypo<strong>the</strong>sis, a SNP discovery study on <strong>the</strong> Sp110 gene for use on a case-control association study has been undertaken comparing 500 healthy and 350 infected Holstein-Friesian cattle. As nucleotide variation <strong>of</strong> bovine Sp110 gene is poorly described, we focused on SNP (single nucleotide polymorphisms) discovery by comparative sequencing. Looking for functional variation, 9980 bp out <strong>of</strong> 51532 bp <strong>of</strong> <strong>the</strong> gene were sequenced in 30 animals (15 healthy / 15 infected). This included <strong>the</strong> 300 bp <strong>of</strong> <strong>the</strong> promoter, 12 exons, and both 3’- and 5’- UTR regions <strong>of</strong> <strong>the</strong> gene. In all, 20 SNPs were detected for <strong>the</strong> first time in Holstein-Friesian cattle. The application <strong>of</strong> <strong>the</strong> selected SNP polymorphisms to <strong>the</strong> case-control samples has not yet revealed any association. #156 Different cytokine responses after interaction between human macrophages and Mycobacterium avium subsp. paratuberculosis type II and type III strains Erika Borrmann, Petra Moebius, Heike Köhler Institute <strong>of</strong> Molecular Pathogenesis, Friedrich-Loeffler-Institute, Jena, Germany Objective: The crucial process in <strong>the</strong> pathogenesis <strong>of</strong> paratuberculosis is <strong>the</strong> internalization <strong>of</strong> Mycobacterium avium subsp. paratuberculosis (MAP) by intestinal host macrophages, its survival and intracellular multiplication. The knowledge about <strong>the</strong>se pathogenic mechanisms is still limited. Fur<strong>the</strong>rmore, despite similarities <strong>of</strong> paratuberculosis to Crohn`s disease (CD) concerning <strong>the</strong>ir histopathology and clinical symptoms, <strong>the</strong> putative pathogenic role <strong>of</strong> MAP in CD is unknown. The objective <strong>of</strong> <strong>the</strong> current study was to investigate differences in <strong>the</strong> cytokine responses <strong>of</strong> human monocytes after interaction with three MAP strains <strong>of</strong> different genotypes. Materials and Methods: The human monocyte cell line THP-1 was incubated with two MAP strains <strong>of</strong> type II (reference strain ATCC19698, one bovine field isolate) and one MAP strain <strong>of</strong> type III (intermediate type) characterized by IS900-RFLP and MIRU-VNTR genotyping before. An in vitro model was established and <strong>the</strong> test conditions were standardized. As parameters for <strong>the</strong> determination <strong>of</strong> <strong>the</strong> impact <strong>of</strong> <strong>the</strong>se strains on <strong>the</strong> macrophage responses, <strong>the</strong> production <strong>of</strong> mRNA <strong>of</strong> <strong>the</strong> cytokines TNF-alpha, IL-1, and IL-10 was determined by quantitative real-time PCR using GAPDH as housekeeping gene. The biological active proteins IL-1 and IL- 10 were measured using ELISA, TNF-alpha by a cytotoxicity test. Results: Differences in <strong>the</strong> expression <strong>of</strong> mRNA and <strong>the</strong> production <strong>of</strong> <strong>the</strong> proteins IL-1, IL-10 and TNFalpha were detected after interaction <strong>of</strong> human monocytes with <strong>the</strong> three MAP-strains. These differences were determined between MAP type III and MAP type II as well as between <strong>the</strong> MAP type II reference and field strains. Conclusion: It seems that MAP strains <strong>of</strong> diverse genotypes induce different responses <strong>of</strong> macrophages. Fur<strong>the</strong>r investigations will show if <strong>the</strong>re is a correlation <strong>of</strong> <strong>the</strong> cytokine responses <strong>of</strong> macrophages and <strong>the</strong> virulence properties <strong>of</strong> <strong>the</strong>se specific MAP strains. 134
Page 1 and 2:
Book of Abstracts
Page 4 and 5:
Proceedings <stron
Page 6 and 7:
Grant Support and Sponsorship <stro
Page 8 and 9:
Scott Wells - Planning Committee Ch
Page 10 and 11:
Monday, August 10, 2009 - All sessi
Page 12 and 13:
Kari Røste Lybeck, Anne Kristine S
Page 14 and 15:
1120-1140 #88: Influence of
Page 16 and 17:
Concurrent Session B - Rm 175 Wille
Page 18 and 19:
Thursday, August 13 Thursday, Augus
Page 20:
Diagnostics and Genotyping Convenor
Page 23 and 24:
#229 Detecting Johne’s Disease he
Page 25 and 26:
Kalis CHJ, Hesselink JW, Barkema HW
Page 27 and 28:
MATERIALS AND METHODS Sampling For
Page 29 and 30:
To better determine the</st
Page 31 and 32:
#107 Multilocus Short Sequence Repe
Page 33 and 34:
Fig. 1. Dendogram (showing genetic
Page 35 and 36:
#105 MIRU-VNTR genotyping o
Page 37:
Diagnostics and Genotyping Poster A
Page 40 and 41:
Missouri). Serum from a cow with do
Page 42 and 43:
#7 Significance of
Page 44 and 45:
RESULTS Of the 327
Page 46 and 47:
Direct fecal nested Map PCR testing
Page 48 and 49:
#16 Histopathological and immunohis
Page 50 and 51:
#19 Development of
Page 52 and 53:
#36 Genetic diversity of</s
Page 54 and 55:
#60 Evaluation of
Page 56 and 57:
#66 Comparison of
Page 58 and 59:
(b) IS900 Nested PCR, using p90-p91
Page 60 and 61:
(d) f57 Real Time PCR. The figure o
Page 62 and 63:
• MIRU (3 loci); • VNTR-MIRU (7
Page 64 and 65:
indexes, compared to each single an
Page 66 and 67:
#108 Evaluation of
Page 68 and 69:
#110 Detection and molecular confir
Page 70 and 71:
endoscopic criteria and histopathol
Page 72 and 73:
Bull TJ, McMinn EJ, Sidi-Boumedine
Page 74 and 75:
#131 Seroprevalence of</str
Page 76 and 77:
could be partly attributed to a sma
Page 78 and 79:
#146 Analysis of v
Page 80 and 81:
#162 Comparison of
Page 82 and 83:
#187 Comparison of
Page 84 and 85: #191 Potentiating day-old blood sam
Page 86 and 87: samples from the s
Page 88 and 89: (MAA) (MAP87 and MAP3776) and 1 fro
Page 90 and 91: #196 Inter- and intra-subtype genot
Page 92 and 93: RESULTS In 601 (29%) of</st
Page 94 and 95: #211 Culture of my
Page 96 and 97: #242 Application and field validati
Page 98 and 99: #251 Diagnosis of
Page 100 and 101: Table 2. Paratuberculosis PCR resul
Page 102 and 103: #254 Programmes on Paratuberculosis
Page 104 and 105: Fig. 1. European countries specifyi
Page 106 and 107: A bulk milk quality assurance progr
Page 108 and 109: Republic of Lithua
Page 110 and 111: #276 Elimination of</strong
Page 112 and 113: other projects and
Page 114 and 115: Woodbine KA, Schukken YH, Green LE,
Page 116 and 117: Host Response and Immunology Keynot
Page 118 and 119: #33 MERKAL AWARD LECTURE No interfe
Page 120 and 121: #75 Interleukin 17 and interleukin
Page 122 and 123: #173 Local and systemic roles for b
Page 124 and 125: #23 Role of Mycoba
Page 126 and 127: Host Response and Immunology Poster
Page 128 and 129: #28 Intestinal MAP Infection via Pe
Page 130 and 131: #73 Age susceptibility of</
Page 132 and 133: #95 Role of nitric
Page 136 and 137: #157 Study of <str
Page 138 and 139: #163 Immunohistochemical expression
Page 140 and 141: #193 Development of</strong
Page 142 and 143: #209 Murine macrophages carrying an
Page 144 and 145: #224 Application of</strong
Page 146 and 147: Table 1 UK and Cyprus MAP survey Re
Page 148 and 149: #228 Mycobacterium avium ssp. parat
Page 150 and 151: #145 MERKAL AWARD LECTURE Multinomi
Page 152 and 153: #50 Assessment of
Page 154 and 155: Table 1. Posterior median (CrIs) <s
Page 156 and 157: #55 Birth clusters of</stro
Page 158 and 159: from their dam, se
Page 160 and 161: #88 Influence of b
Page 162 and 163: #104 Relation between faecal shedde
Page 164 and 165: Epidemiology Poster Abstracts 109 1
Page 166 and 167: #22 The Prevalence of</stro
Page 168 and 169: CONCLUSION IS1311-based nested Ma/M
Page 170 and 171: #42 Seroprevalence survey o
Page 172 and 173: #77 Characterisation of</st
Page 174 and 175: Environmental samples were collecte
Page 176 and 177: REFERENCES 1) Chiodini RJ, Van Krui
Page 178 and 179: #113 Effect of soi
Page 180 and 181: #136 Error-adjusted, variance parti
Page 182 and 183: Latium Region Viterbo districts RES
Page 184 and 185:
#144 The suitability of</st
Page 186 and 187:
#178 Age structure of</stro
Page 188 and 189:
A statistically significant lower f
Page 190 and 191:
#183 Sero-prevalence of</st
Page 192 and 193:
#218 Johne’s disease in t
Page 194 and 195:
#244 A survey to estimate t
Page 196 and 197:
Control Programs Keynote Lecture Re
Page 198 and 199:
#202 Milk quality assurance for par
Page 200 and 201:
#141 Control of pa
Page 202 and 203:
#198 Use of small
Page 204 and 205:
Control Programs Poster Abstracts 1
Page 206 and 207:
RESULTS Loss of al
Page 208 and 209:
#40 Managing Bovine Johnes Disease
Page 210 and 211:
#74 Economic analysis of</s
Page 212 and 213:
#87 Evaluation of
Page 214 and 215:
separately for each existing herd <
Page 216 and 217:
#98 An inter-laboratory ring-trial
Page 218 and 219:
#120 Paratuberculosis vaccine inter
Page 220 and 221:
The statement provides: • A stand
Page 222 and 223:
COMPLEMENTARY ASPECTS On-Farm Disea
Page 224 and 225:
#168 Paratuberculosis in Iran: past
Page 226 and 227:
#180 Behavioural change in owners <
Page 228 and 229:
the fact that <str
Page 230 and 231:
#182 An integrated risk based appro
Page 232 and 233:
etween 0 to 10, depending on <stron
Page 234 and 235:
#203 Milk quality assurance for par
Page 236 and 237:
#234 Johne’s disease vaccine: a c
Page 238 and 239:
Pathogenomics and MAP Biology Conve
Page 240 and 241:
#45 Identification of</stro
Page 242 and 243:
#117 Adsorption of
Page 244 and 245:
#222 Atypical structural features <
Page 246 and 247:
Pathogenomics and MAP Biology Persp
Page 248 and 249:
247
Page 250 and 251:
#52 Is ‘Indian Bison Type’ Myco
Page 252 and 253:
#69 In vitro susceptibility <strong
Page 254 and 255:
#118 Cloning, expression and purifi
Page 256 and 257:
#176 A reference proteomic pr<stron
Page 258 and 259:
#213 Construction the</stro
Page 260 and 261:
#230 Iron concentration dependent s
Page 262 and 263:
261
Page 264 and 265:
263
Page 266 and 267:
Abstract not available at press tim
Page 268 and 269:
#174 Sharing of
Page 270 and 271:
#177 Associations between CARD15 po
Page 272 and 273:
Perspectives Talk: Public Health My
Page 274 and 275:
273
Page 276 and 277:
A total of 206 qua
Page 278 and 279:
Feller, M., K. Huwiler, R. Stephan,
Page 280 and 281:
In 2008, the Ameri
Page 282 and 283:
#115 Crohn’s disease and ruminant
Page 284 and 285:
International John
Page 286 and 287:
#76 Towards improved reporting stan
Page 288 and 289:
#97 The EU ParaTBTools Project - Th
Page 290 and 291:
#241 US Vaccine Initiative through
Page 292 and 293:
Paratuberculosis ELISA Reliable, si
Page 294 and 295:
PRIONICS AG WAGISTRASSE 27A PHONE +
show all

Proceedings of the 10th International Colloquium on Paratuberculosis

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?