Proceedings of the 10th International Colloquium on Paratuberculosis

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#75 Interleukin 17 and interleukin 23 gene expression differentiate severe pathology in Johne’s disease Mark William Robinson, Rory O’Brien, Frank Griffin University <strong>of</strong> Otago, New Zealand The gastrointestinal immune response to infectious agents such as Mycobacterium avium subspecies paratuberculosis must be balanced in order to effectively contain <strong>the</strong> infection while at <strong>the</strong> same time avoiding excessive tissue destruction. As such, differences in <strong>the</strong> host immune response to challenge can potentially influence disease progression and outcome <strong>of</strong> Johne’s disease. Using an experimental infection model <strong>of</strong> Johne’s disease in red deer (Cervus elaphus) we studied <strong>the</strong> immune responses during different pathological states. A group <strong>of</strong> 30 red deer were challenged with ~10 9 colony forming units <strong>of</strong> bovine strain Mycobacterium avium subspecies paratuberculosis and were monitored for Johne’s disease development over <strong>the</strong> course <strong>of</strong> 1 year. Peripheral blood samples were acquired at 20 weeks post-infection and post-jejunal lymph node (PJJLN) samples following necropsy at ei<strong>the</strong>r 20-28 weeks post-infection for clinically diseased animals or at 50 weeks post-infection for remaining animals. Animals were retrospectively grouped into 3 pathological outcomes (Infected, Paucibacillary Diseased or Multibacillary Diseased) on <strong>the</strong> basis <strong>of</strong> histology scores and tissue culture results. Gene expression <strong>of</strong> transcripts for IFNg, IL2, Tbet, IL1a, TNFa, TRAF1, GATA3, IL4, Foxp3, IL10, IL17, IL23p19, IL6 and TGFb were detected using quantitative real time PCR and data was analysed using REST s<strong>of</strong>tware to determine significant changes in gene expression levels. Stimulation <strong>of</strong> peripheral blood mononuclear cells gave distinctive patterns <strong>of</strong> gene expression for all 3 groups with strong Type 1 responses in both <strong>the</strong> Infected and Paucibacillary Diseased groups while <strong>the</strong> Multibacillary Diseased group showed a mixed immune response with significant up-regulation <strong>of</strong> <strong>the</strong> IL23p19 gene transcript. The PJJLN gene expression pr<strong>of</strong>iles <strong>of</strong> <strong>the</strong> Multibacillary Diseased group exhibited a highly inflammatory pr<strong>of</strong>ile with no evidence <strong>of</strong> a role played by T regulatory or Type 2 immune responses. Gene expression pr<strong>of</strong>iles provide novel diagnostic markers for different pathological states <strong>of</strong> Johne’s disease and support <strong>the</strong> hypo<strong>the</strong>sis that differing host immune responses direct disease progression and pathological outcome. 119
#119 The early IL-10 response in ovine Johne’s disease Kumudika de Silva, Douglas Begg, Deborah Taylor, Richard Whittington University <strong>of</strong> Sydney, Australia The classic early immunological response to Mptb is mediated by T-cells with predominant interferon gamma activity. This response is curbed as clinical signs <strong>of</strong> disease appear and may, at least partly, be due to a surge in <strong>the</strong> activity <strong>of</strong> <strong>the</strong> immunosuppressive cytokine interleukin 10 (IL-10). While <strong>the</strong>re is evidence <strong>of</strong> IL-10 activity in <strong>the</strong> later stages <strong>of</strong> disease its involvement at earlier time points is not known. The aim <strong>of</strong> this study was to monitor <strong>the</strong> antigen-specific IL-10 response during experimental Johne’s disease. Merino lambs were left unchallenged (controls, n=20) or challenged orally with Mptb (n=40). Lymphocytes isolated from blood (every 4 months) and lymph nodes (11-13 months after challenge) were cultured in <strong>the</strong> presence or absence <strong>of</strong> Mptb antigen. IL-10 in culture supernatants was determined by ELISA. Faecal shedding and presence <strong>of</strong> Mptb in tissue samples were determined by culture in a radiometric system (Bactec). Histological lesion type was also assessed. Antigen-specific IL-10 secretion in peripheral blood <strong>of</strong> sheep exposed to Mptb was significantly higher than in control animals (P < 0.001) as early as 4 months p.i. and increased progressively. The peripheral blood IL-10 response in animals with multibacillary lesions tended to differ from animals with no histological lesions or with paucibacillary lesions. In ileal and jejunal lymph node cells, IL-10 secretion was significantly higher in <strong>the</strong> exposed animals (P < 0.05). In addition, IL-10 secretion from <strong>the</strong>se cells was significantly higher in animals with no lesions or with paucibacillary lesions when compared to animals with multibacillary lesions. Our novel findings demonstrate that IL-10 activity occurs soon after exposure to Mptb and may play a role in determining disease outcome. 120
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Book of Abstracts
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Proceedings <stron
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Grant Support and Sponsorship <stro
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Scott Wells - Planning Committee Ch
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Monday, August 10, 2009 - All sessi
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Kari Røste Lybeck, Anne Kristine S
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1120-1140 #88: Influence of
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Concurrent Session B - Rm 175 Wille
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Thursday, August 13 Thursday, Augus
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Diagnostics and Genotyping Convenor
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#229 Detecting Johne’s Disease he
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Kalis CHJ, Hesselink JW, Barkema HW
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MATERIALS AND METHODS Sampling For
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To better determine the</st
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#107 Multilocus Short Sequence Repe
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Fig. 1. Dendogram (showing genetic
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#105 MIRU-VNTR genotyping o
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Diagnostics and Genotyping Poster A
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Missouri). Serum from a cow with do
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#7 Significance of
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RESULTS Of the 327
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Direct fecal nested Map PCR testing
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#16 Histopathological and immunohis
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#19 Development of
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#36 Genetic diversity of</s
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#60 Evaluation of
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#66 Comparison of
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(b) IS900 Nested PCR, using p90-p91
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(d) f57 Real Time PCR. The figure o
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• MIRU (3 loci); • VNTR-MIRU (7
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indexes, compared to each single an
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#108 Evaluation of
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#110 Detection and molecular confir
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Page 74 and 75: #131 Seroprevalence of</str
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Page 78 and 79: #146 Analysis of v
Page 80 and 81: #162 Comparison of
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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
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Page 110 and 111: #276 Elimination of</strong
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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
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Page 124 and 125: #23 Role of Mycoba
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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
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Page 138 and 139: #163 Immunohistochemical expression
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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
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Page 154 and 155: Table 1. Posterior median (CrIs) <s
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Page 168 and 169: CONCLUSION IS1311-based nested Ma/M
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#42 Seroprevalence survey o
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#77 Characterisation of</st
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Environmental samples were collecte
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REFERENCES 1) Chiodini RJ, Van Krui
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#113 Effect of soi
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#136 Error-adjusted, variance parti
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Latium Region Viterbo districts RES
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#144 The suitability of</st
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#178 Age structure of</stro
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A statistically significant lower f
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#183 Sero-prevalence of</st
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#218 Johne’s disease in t
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#244 A survey to estimate t
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Control Programs Keynote Lecture Re
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#202 Milk quality assurance for par
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#141 Control of pa
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#198 Use of small
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Control Programs Poster Abstracts 1
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RESULTS Loss of al
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#40 Managing Bovine Johnes Disease
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#74 Economic analysis of</s
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#87 Evaluation of
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separately for each existing herd <
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#98 An inter-laboratory ring-trial
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#120 Paratuberculosis vaccine inter
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The statement provides: • A stand
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COMPLEMENTARY ASPECTS On-Farm Disea
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#168 Paratuberculosis in Iran: past
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#180 Behavioural change in owners <
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the fact that <str
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#182 An integrated risk based appro
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etween 0 to 10, depending on <stron
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#203 Milk quality assurance for par
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#234 Johne’s disease vaccine: a c
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Pathogenomics and MAP Biology Conve
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#45 Identification of</stro
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#117 Adsorption of
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#222 Atypical structural features <
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Pathogenomics and MAP Biology Persp
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#52 Is ‘Indian Bison Type’ Myco
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#69 In vitro susceptibility <strong
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#118 Cloning, expression and purifi
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#176 A reference proteomic pr<stron
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#213 Construction the</stro
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#230 Iron concentration dependent s
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Abstract not available at press tim
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#174 Sharing of
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#177 Associations between CARD15 po
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Perspectives Talk: Public Health My
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A total of 206 qua
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Feller, M., K. Huwiler, R. Stephan,
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In 2008, the Ameri
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#115 Crohn’s disease and ruminant
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International John
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#76 Towards improved reporting stan
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#97 The EU ParaTBTools Project - Th
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#241 US Vaccine Initiative through
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Paratuberculosis ELISA Reliable, si
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PRIONICS AG WAGISTRASSE 27A PHONE +
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