Progress 10/01/07 to 09/30/12
Outputs
OUTPUTS: 1) Campylobacter jejuni isolates from calves have A, B and C lipooligosaccharide (LOS) biosynthetic locus classes similar to human Guillain Barre syndrome (GBS) associated strains C. jejuni is a frequent cause of gastroenteritis and can trigger the acute neuropathy GBS. A family managing a Midwest dairy operation reported a history of C. jejuni infections that lasted several days and recurred. Because 37.7% of dairy cattle shed C. jejuni, we sought to determine whether calves were the source of family infections. We cultured fecal samples from 25 calves, 1 dog, and 1 family member for C. jejuni. Isolates were characterized for LOS biosynthesis loci and by multi-locus sequence typing. C. jejuni was cultured from 15 (60%) of calves and one asymptomatic family member; the dog was negative. C. coli was cultured from 2 (1%) of calves. Several calf C. jejuni isolates had LOS classes A, B and C, strongly associated with GBS. The human isolate had an LOS class E, associated with enteric disease. 3 calves had C. jejuni with LOS class E. Typing results suggest that some calf and human C. jejuni isolates have similar characteristics. Multiple LOS classes in isolates from the calves demonstrate that there are diverse C. jejuni populations on this farm. 2) Comparison of induced animal models for GBS after C. jejuni infection. Mice were infected with C. jejuni strains HB93-13 and 260.94, from GBS patients and examined daily for clinical signs of campylobacterosis and tested weekly for neurological phenotypes. Autoantibodies to GM1 and GD2a gangliosides in serum were assayed by ELISA and nerves were removed at necropsy, fixed, and stained to detect nerve lesions and myelin damage. Antiganglioside antibodies and neurological disease develop in NOD WT, NOD B7-2-/-, and NOD IL-10-/- mice subsequent to C. jejuni infection with GBS strains. Mice developed neurological signs including decreased activity, failure to rear, splayed feet, shortened stride length, foot drag, and hind limb paralysis among other signs. Sciatic nerve lesions were consistent with acute inflammatory demyelinating polyradiculoneuropathy (AIDP). 3) C. jejuni induces mixed Type 1 and 17 responses in a mouse model of colitis. C57BL/6 IL-10+/+ and congenic IL-10−/− mice serve as C. jejuni colonization and colitis models. We hypothesized that in the absence of the regulatory cytokine IL-10, uncontrolled induction of proinflammatory cytokines and infiltration of inflammatory cells is responsible for tissue obliteration in colitis. Colon, lymph nodes and plasma cytokine analysis revealed significant time-dependent up regulation of IFNgamma, IL-17A, IL-6, TNFalpha and IL-1beta. Colon flow cytometry and immunohistochemistry revealed time-dependent increases in numbers of infiltrating macrophages, neutrophils, T cell subsets, NK cells and Thy1+ CD3- innate lymphocytes. Intra-cellular cytokine staining of colon and lymph node lymphocytes further linked IFNgamma, IL-17 and IL-22 upregulation to specific T cell subsets and innate lymphocytes. Time-dependent up regulation in anti-C. jejuni reactive IgG2b, IgG2c and IgG3, but not IgG1 or IgM were seen in infected mouse plasma. PARTICIPANTS: Several students were mentored for study of enteric diseases of food animals. John Paul Jerome (PhD candidate) successfully defended his PhD thesis and has gone on to a job as a postdoc with Professor Sheng Yang He who has an endowed chair to study food borne pathogens in plants. Four other students are pursuing PhD degrees in food safety related to these enteric pathogens including Jessica St. Charles, Ankit Malik, Barbie Gadsden, DVM, and Phillip Brooks. Two of these students have passed their preliminary exams. Dr. Gadsden is preparing for this exam. Two of the graduate students are underrepresented minorities (African American). Four undergraduate students are working in the lab on these projects including Hahyung Kim, Elizabeth Gensterbloom, Julian Yu, and Anthony Brooks. Anthony was accepted to medical school and will start in 2013. Dr. Mansfield organized a seminar in Food and Waterborne Diseases for the faculty and students of Michigan State University on October 19, 2012. People attending came from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. The speakers included Shannon D. Manning, PhD, MPH Michigan State University (Evolution and Pathogenesis of E. coli O104:H4 infections) and Kathryn Eaton DVM, PhD, University of Michigan (Host, pathogen, and microbiota contribute to disease severity in EHEC-infected mice). Dr. Mansfield helped to organize and attended the USDA Enteric Diseases Meeting NC1041 in Chicago, Illinois on December 1st and 2nd as part of the Conference of Research Workers on Animal Diseases. Her graduate student Jessica St. Charles gave a talk at CRWAD about her work entitled "Campylobacter jejuni isolates from calves have A, B and C lipooligosaccharide (LOS) biosynthetic locus classes similar to human Guillain Barre syndrome associated strains". For the same CRWAD meeting, Dr. Mansfield gave a talk at CRWAD entitled "Comparison of induced small animal models for Guillain Barre syndrome (GBS) as post infectious sequelae to Campylobacter jejuni infection". Dr. Mansfield gave an invited talk entitled "Murine models of the autoimmune neuropathy Guillain Barre Syndrome" for the Small Animal Models of Enteric Diseases, NIAID, National Institutes of Health, September 13-14, 2012. She also gave an invited talk entitled "Campylobacter jejuni induces mixed Type 1 and 17 responses in acute and chronic disease" for the Enterics Research Investigational Network, Cooperative Research Centers Annual Meeting, National Institutes of Health, Seattle, Washington, May 22-23, 2012. TARGET AUDIENCES: The main target audience is the scientific community studying food safety and medical interventions for food borne pathogens. We reach this audience with high quality research publications in scientific journals as well as the talks to scientific meetings and producer groups for food animals. We also have a long term affect by teaching undergraduate students, graduate students and postdoctoral candidates these advances and strategies for studying food borne pathogens. The ultimate goals of this work are to translate this information into preventatives and treatments for food borne diseases. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Study #1 Impact: This study demonstrates for the first time that calves colonized with C. jejuni without disease can carry strains that have the genetic characteristics associated with those from GBS patients. This implicates calves as a source for the strains of campylobacter that induce autoimmune diseases like this peripheral neuropathy. These are the most dangerous forms of this food borne bacterium. Study #2 Impact: NOD, NOD B7-2-/-, and NOD IL-10-/- mice can serve as models for autoimmune neuropathies in humans. New models for GBS can be used to test treatment and prevention modalities and therapies. Study #3 Impact: Data demonstrate a mixed Type 1 and Type 17 induced colitis in C. jejuni 11168 challenged C57BL/6 IL-10-/- mice. The work shows for the first time the adaptive immune response that leads to inflammatory bowel disease in this genetically susceptible mouse model. These results can be used to understand this process in humans.
Publications
- Bell JA, Jerome JP, Plovanich-Jones AE, Smith EJ, Gettings JR, Kim HY, Landgraf JR, Lefebure T, Kopper JJ, Rathinam VA, St Charles JL, Buffa BA, Brooks AP, Poe SA, Eaton KA, Stanhope MJ, Mansfield LS. 2012. Outcome of infection of C57BL/6 IL-10(-/-) mice with Campylobacter jejuni strains is correlated with genome content of open reading frames up- and down-regulated in vivo. Microbial Pathogenesis 2012 Aug 31. pii: S0882-4010(12)00151-9. doi: 10.1016/j.micpath.2012.08.001.
- Kim JS, Artymovich KA, Hall DF, Smith EJ, Fulton R, Bell JA, Dybas L, Mansfield LS, Tempelman R, Wilson DL, Linz JE. 2012. Passage of Campylobacter jejuni through the chicken reservoir or mice promotes phase variation in contingency genes Cj0045 and Cj0170 that strongly associates with colonization and disease in a mouse model. Microbiology February 16, 2012 as doi:10.1099/mic.0.057158-0.
|
Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: 1)Campylobacter jejuni: What's in a strain The presence of hypermutable homopolymeric DNA associated with contingency loci is a striking part of the C. jejuni genome. Using a mouse model of campylobacteriosis we showed that C. jejuni NCTC11168 re-isolated from infected mice (mouse-adapted) is more virulent than the ancestral inoculum. Mouse-adapted C. jejuni also had altered ability to autoagglutinate and interact with epithelial cells. After genetic analysis using Illumina genome re-sequencing, we showed that the only difference between mouse-adapted and ancestral C. jejuni was in homopolymeric guanine tracts of 13/29 contingency loci. This suggests that contingency loci play a role in pathogenesis. We then tested the effect of bottlenecking contingency loci mutational diversity on C. jejuni virulence phenotypes. A variant subjected to a single colony bottleneck could not colonize mice. Considering the tract length diversity found in 29 hypermutable loci, a single strain of C. jejuni potentially contains 536 million distinct genotypes. This variation is subject to selective pressure and can affect contingency genes that alter autoagglutination, epithelial cell interactions, fitness/virulence in a mouse model, motility, and antigenicity. The ubiquity of contingency loci mutations may confound analysis of C. jejuni by traditional molecular cloning methods that involve single-cell bottlenecks. 2)C. jejuni NCTC 11168 induces a mixed Type1 and Type17 cytokine and cellular response in IL-10knockout mice. C. jejuni is the leading cause of foodborne enteritis. C57BL/6 IL-10+/+ and congenic IL-10−/− mice serve as C. jejuni colonization and colitis models. IL-10-/- mice orally inoculated with C. jejuni develop colitis characterized by mucosal edema, crypt abscesses, erosions or ulcerations of the epithelium, and extensive infiltration of colon tissues with polymorphonuclear and mononuclear cells in a time-dependent but dose-independent manner. We hypothesized that in the absence of the regulatory cytokine IL-10, uncontrolled induction of proinflammatory cytokines and infiltration of inflammatory cells is responsible for tissue obliteration. We analyzed cytokines and cell types in colon and mesenteric lymph nodes, and C. jejuni specific antibody and cytokine response in serum. Colon homogenate and serum ELISA revealed significant up regulation of IFNgamma, IL-17A, IL-22, IL-6 and IL-1beta but not IL-4. Colon flow cytometric analysis revealed time-dependent increases in number of infiltrating myeloid cells, neutrophils, T cells and innate lymphocytes. Mesenteric lymph node analysis revealed increases in IFNgamma positive cells in the CD4+, CD8+ and gammadelta T cell and NK cell compartments. IL-17 positive cells were increased in CD4+ T cell and Thy1hiCD3- innate lymphocyte compartments. IL-22 positive cells were increased from CD4+ T cell and NK cell compartments. Time-dependent up regulation in C. jejuni specific IgG2b (Th17), IgG2c and IgG3 (Th1), but not IgG1 (Th2) or IgM were observed in the serum of infected mice. Data demonstrate a mixed Th1 and Th17 mediated colitis in C. jejuni challenged C57BL/6 IL-10-/- mice. PARTICIPANTS: Principal Investigator: Linda S. Mansfield (Station Representative), Julia A. Bell, Ph.D., Research Assistant Professor; Graduate Students: John Paul Jerome, Ph. D. candidate, Jessica L. St. Charles, Ph. D. candidate, Ankit Malik, Ph. D. candidate, Barbie Gadsden, DVM, Ph. D. candidate TARGET AUDIENCES: The target audience for this work is mainly scientists in the many related fields. Students studying to gain expertise in this field are an important target audience for the work. Graduate students: Once again, several students were mentored for study of enteric diseases of food animals. Jamie Jennifer Kopper (DVM candidate) successfully defended her PhD thesis and will go on to finish two more years of veterinary school. Four other students are pursuing PhD degrees in food safety related to these enteric pathogens including John Paul Jerome, Jessica St. Charles, Ankit Malik and Barbie Gadsden, DVM. Three of these students have passed their preliminary exams, two during 2011. One is preparing for this exam. Four undergraduate students are working in the lab on these projects. One undergraduate was accepted to and started medical school during 2011. Seminars to scientists: Dr. Mansfield organized a Spring seminar in Food and Waterborne Diseases for the faculty and students of Michigan State University. People attending have come from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. The speakers included Christopher Waters, PhD (Biofilms in foodborne bacteria) and John Paul Jerome (Contingency genes in Campylobacter jejuni). Also, Dr. Mansfield attended and presented at a scientific conference held by the National Institutes of Health in Baltimore, Maryland on Enteric Diseases. Dr. Mansfield helped to organize and attended the USDA Rushmore Enteric Diseases Meeting NC1041 in Chicago, Illinois on December 3rd and 4th. Her graduate student John Paul Jerome gave a talk about his work entitled "Contingency loci dynamics during Campylobacter jejuni infection". Dr. Mansfield gave a talk for the Department of Large Animal Clinical Sciences at Michigan State University entitled "Investigating the role of Campylobacter jejuni in acute and chronic disease: the genetic basis of pathotypes" on 02/01/2011. PROJECT MODIFICATIONS: none
Impacts
These findings provide cost effective animal models that can be used to study the genetic basis of virulence of the enteric pathogen C. jejuni. Furthermore, in this study period we used these murine models to determine several genetic determinants of the host that control susceptibility to enteric disease and the various pathotypes observed in patients due to this bacterium. We now understand that contingency genes are virulence factors in C. jejuni. Virulence Study Impact: We showed that C. jejuni strains possess hypermutable loci that undergo slip strand mutagenesis with a single passage in the host. These homopolymeric tracts occur mainly in genes encoding the surface structure of the bacterium. This has great implications for developing vaccines against C. jejuni. It is highly likely that this bacterium exists as a populations of "strains" that evolve very rapidly once a selective pressure is applied. This means that the current strategy for having a single "clone" of the bacterium act as a vaccine will certainly fail as other members of the population take over. It will be very important to examine this process fully to understand how to circumvent this adaptive strategy of the bacterium when designing vaccines. We also studied adaptive immune responses to C. jejuni especially those associated with development of colitis.
Publications
- Bell JA, Jerome JP, Plovanich-Jones AE, Smith EJ, Gettings JR, Kim HY, Landgraff JR, Lefebure T, Kopper JJ, Rathinam VA, St. Charles JL, Buffa BA, Brooks AP, Poe SA, Eaton KA, Stanhope MJ, Mansfield LS. 2011. A comparative genomics approach to the C. jejuni virulome: Campylobacter jejuni virulence phenotypes (pathotypes), genotypes, and gene expression in C57BL/6 IL-10-/- mice.
- Joo-Sung Kim, Katherine A. Artymovich, David F. Hall, Julia A. Bell, Leslie Dybas, Eric J. Smith, Linda S. Mansfield, and John E. Linz. 2011. Phase variation in Campylobacter jejuni contingency genes Cj0045 and Cj0170 is associated with virulence in a mouse model for human disease.
- J. L. St. Charles, A. M. Zaleski, E. J. Smith, J. A. Bell, L. S. Mansfield, 2011. Examination of Miller Fisher Syndrome in a Murine Model, Campylobacter, Helicobacter and Related Organisms, Vancouver, British Columbia, August 28-September 1, 2011.
- L. S. Mansfield, 2011. Overview of Emerging Campylobacter Species, Campylobacter, Helicobacter and Related Organisms, Vancouver, British Columbia, August 28-September 1, 2011.
- John P. Jerome, J.A. Bell, J.E. Barrick, C.T. Brown, Linda S. Mansfield. 2011. Campylobacter jejuni: What is in a strain That which is not a clone, Campylobacter, Helicobacter and Related Organisms, Vancouver, British Columbia, August 28-September 1, 2011.
- John P. Jerome, J.A. Bell, J.E. Barrick, C.T. Brown, Linda S. Mansfield, 2011. Contingency loci dynamics during Campylobacter jejuni infection, The Fourth International Rushmore Meeting, Chicago, IL, December 3-4, 2011.
- A. Malik, H. Kim and L.S. Mansfield, 2011. Campylobacter jejuni NCTC 11168 induces a mixed Type1 and Type17 cytokine and cellular response in IL-10 deficient murine host, Campylobacter, Helicobacter and Related Organisms, Vancouver, British Columbia, August 28-September 1, 2011.
- Jerome, JP, Bell JA, Plovanich-Jones AE, Barrick JE, Brown CT, Mansfield LS. 2010. Standing genetic variation in contingency genes drives the adaptation of Campylobacter jejuni to a novel host. PLoS ONE 6(1):16399.
|
Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Monitoring Campylobacter jejuni using Fluorescent in situ Hybridization. Background: Campylobacter jejuni is an important cause of food borne enteritis. We have recently developed a murine model for gastroenteritis and systemic disease using C3Bir tlr knockout IL 10 knockout mice with C. jejuni 11168. These mice exhibited extraintestinal spread, demonstrated by enlarged spleens and gastrointestinal (GI) lymph nodes and positive blood cultures. Therefore, they can serve as surrogates for immunocompromised patients to further understanding of the pathogenesis of C. jejuni that escapes the bowel. We hypothesized that C. jejuni can be detected in GI lumen, GI tissues and extraintestinal tissues of C3Birtlr KO IL 10 KO mice using fluorescent imaging. Methods: Our objective was to determine organ specific recovery of C. jejuni in this murine model post-challenge. We developed a fluorescence in situ hybridization (FISH) method to detect C. jejuni in mouse tissues. This method utilized a C. jejuni specific, fluorescently labeled oligonucleotide probe, which is hybridized to sections of relevant tissue. We then tested whether UV pre-bleaching of these paraffin-embedded tissues would reduce autofluorescence. After hybridization the tissue sections were scored in a blinded fashion and the results indicated that FISH may be an efficient way to monitor for C. jejuni invasion. Results: FISH was used following pre-bleaching to detect C. jejuni in paraffin-embedded tissues. A scoring system was created to assess the specificity of the FISH staining. When scored in a blinded fashion, the total score of the positive slides was 97 of a possible 220 points, while the total score of the negative slides was 25 of 220. The average score of a positive slide was 19.4 of a possible 44 points, while negative slides averaged a score of 5 of 44. Prebleaching of autofluorescent epitopes in tissues significantly enhanced the ability to visualize this bacterium in tissues after challenge infection. Conclusion: Since all slides were blinded to the scorer, the results showed that FISH can be a reliable method of tracking C. jejuni in paraffin-embedded tissue. PARTICIPANTS: Collaborators on the project were Michael Stanhope (Cornell University) and Michael Konkel (Washington State University). TARGET AUDIENCES: Focus on disseminating knowledge - Provide training or continuing education opportunities and dissemination of information to students, producers, veterinarians, and diagnostic laboratories. Several students were mentored for study of enteric diseases of food animals. Five students are pursuing PhD degrees in food safety related to these enteric pathogens including John Paul Jerome, Jessica St. Charles, Jamie Jennifer Kopper (DVM candidate), Ankit Malik and Barbie Gadsden, DVM. Three of these students have passed their preliminary exams. Two are preparing for this exam. Vijay Rathinam, successfully defended his Ph.D. thesis and obtained a postdoctoral position at the University of Massachusetts at Wooster, MA where he is studying the pathogenesis of inflammatory bowel disease due to bacterial pathogens. One undergraduate student working in the lab on these projects was accepted into veterinary schools (Alexander Adrian, University of Minnesota). Dr. Mansfield organized a Fall seminar in Food and Waterborne Diseases for the faculty and students of Michigan State University. People attending have come from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. The speakers included Shannon Manning, PhD, MPH (Shigatoxin producing E. coli) and Robert Britton, PhD (Probiotics for treating enteric diseases). Also, Dr. Mansfield attended and presented at a scientific conference held by the National Institutes of Health at Cambridge, Maryland on Food and Water borne Pathogens. Dr. Mansfield was an invited speaker for the Norman E. Borlaug International Agricultural Science and Technology Fellowship Program mentor at Polish Food Safety Meeting on 04/23/2010 and gave an invited keynote address at the American Society for Microbiology entitled "Examining the genetic basis of pathotypes in Campylobacter jejuni using murine models" on 05/28/2010. Dr. Mansfield organized and attended the USDA Multistate Research Project 2009 Meeting NC1041 in Chicago, Illinois on December 4th and 5th. PROJECT MODIFICATIONS: Our future goals are to define C. jejuni gene expression controlling the different pathotypes that we have defined in the reported murine models. We will continue this study on the genetic diversity of human and animal isolates of C. jejuni that we have acquired through the US Centers for Diseases Control and Prevention and Dr. Al Lastovica, Bellville, South Africa. Methods will include microarray (using a supplemented whole open reading frame (ORF) array from the 7 sequenced strains of C. jejuni), Illumina sequencing, in vivo expression technology (IVET), and selected gene knockouts followed by challenge in the murine models.
Impacts
Study Impact: This method was used to definitively localize C. jejuni in host tissues after an oral challenge infection. The scoring protocol provides semiquantitative measures of colonization and invasion by the bacterium. This method can be used on paraffin embedded tissues providing an easy means of assessing tissue distribution of C. jejuni in samples in long term storage.
Publications
- J.P. Jerome, A.E. Plovanich-Jones, J.A. Bell, Linda S. Mansfield. 2009. Genetic changes during serial passage of Campylobacter jejuni in C57BL/6 IL-10 KO mice are associated with enhanced virulence, Phi Zeta Research Day, College of Veterinary Medicine, Michigan State University, East Lansing, MI.
- Bianca A. Buffa, Julia A. Bell, Vijay A. K. Rathinam, Jamie J. Kopper, Alice J. Murphy, and Linda S. Mansfield. 2009. Responses of C57BL/6 interleukin-10 deficient and proficient mice to infection with low doses of Campylobacter jejuni 11168. Phi Zeta Research Day, College of Veterinary Medicine, Michigan State University, East Lansing, MI.
- Vijay A.K. Rathinam, Daniel M. Appledorn, Kathleen A. Hoag, Andrea Amalfitano and Linda S. Mansfield. 2009. TLR4-MyD88 and TLR4-TRIF signaling axes cooperate in activating dendritic cells in response to an intracellular enteric pathogen. Pattern Recognition Molecules and Immune Sensors, Keystone Meetings, Banff, Alberta CA, March 2009.
- J.A. Bell, J.L. St. Charles, A.J. Murphy, V.A. Rathinam, A.E. Plovanich-Jones, E.L. Stanley, J.E. Wolf, J.R. Gettings, T.S. Whittam, Tristan Lefebure, Michael Stanhope, L.S. Mansfield. 2009. Pathogenicity of Campylobacter jejuni strains in C57BL/6 IL-10 KO mice varies with strain genetic background, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- J.P. Jerome, A.E. Plovanich-Jones, J.A. Bell, Linda S. Mansfield. 2009. Genetic changes during serial passage of Campylobacter jejuni in C57BL/6 IL-10 KO mice are associated with enhanced virulence, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- J.D. Olmstead, V.A.K. Rathinam, L.S. Mansfield. 2009. Determining organ specific recovery of Campylobacter jejuni in a murine disease model, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- J.L. St. Charles, J.A. Bell, and L.S. Mansfield. 2009. Development of murine models for study of Guillain-Barre Syndrome, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- L. S. Mansfield. Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10 KO mice, NC1041 Enteric Diseases of Swine and Cattle: Prevention, Control and Food Safety Meeting, Chicago, IL; December 5-6, 2009.
- L. S. Mansfield. 2010. Developing murine models for the study of enteric pathotypes of Campylobacter jejuni. Pathology Seminar Series, Department of Pathology and Laboratory Medicine, University of Wisconsin, March 17, 2010.
- L.S. Mansfield, J.A. Bell, J.L. St. Charles, J.P. Jerome, A.E. Plovanich-Jones. 2010. Examining the genetic basis of pathotypes in Campylobacter jejuni using murine models, National Veterinary Institute (PIWet), Lublin, Poland, April 19-23, 2010.
- L. S. Mansfield, J. A. Bell, J. L. St. Charles, J. P. Jerome. 2010. Examining the Genetic Basis of Pathotypes in Campylobacter jejuni Using Murine Models, American Society for Microbiology General Meeting, San Diego, CA. May 23-27, 2010.
- J.A. Bell, J. R. Gettings, J. P. Jerome, J. J. Kopper, A. Plovanich-Jones, V. A. K. Rathinam, J. L. St. Charles, E.J. Smith, A. G. Staunton, L. S. Mansfield. 2010. Outcome of Campylobacter jejuni Infection of C57BL/6 IL-10-/- Mice Varies with Strain, American Society for Microbiology General Meeting, San Diego, CA. May 23-27, 2010.
- E. J. Smith, J.D. Olmstead, L. S. Mansfield. 2010 Monitoring Campylobacter jejuni using Fluorescent in situ Hybridization, American Society for Microbiology General Meeting, San Diego, CA. May 23-27, 2010.
- J.P. Jerome, J.A. Bell, A.E. Plovanich-Jones, C.T. Brown, L.S. Mansfield. 2010. The Role of Contingency Genes in the Adaptation of Campylobacter jejuni to a Novel Host, American Society for Microbiology General Meeting, San Diego, CA. May 23-27, 2010.
- J. L. St. Charles, J. A. Bell, L. S. Mansfield. 2010. Development of Murine Model to Study Guillain Barre Syndrome Following Campylobacter jejuni Infection, American Society for Microbiology General Meeting, San Diego, CA. May 23-27, 2010.
- J. St. Charles, J. A. Bell, A. Brooks, B. A. Buffa, J.P. Jerome, J. J. Kopper, V. A. Rathinam, E. Smith, A. Staunton, L. S. Mansfield. Murine Models for Study of Guillain Barre Syndrome (GBS) and Other Autoimmune Neuropathies following Campylobacter infection, National Institutes of Health, Food and Water borne Diseases Integrated Research Network, Cambridge. MD, June 1-4, 2010.
- J. A. Bell, A. Brooks, B. A. Buffa, J. R. Gettings, J.P. Jerome, J. J. Kopper, T. Lefebure, A. E. Plovanich-Jones, V. A. Rathinam, J. St. Charles, E. Smith, A. Staunton, M. J. Stanhope, L. S. Mansfield. 2010. Phylogeny, gene content, and virulence phenotypes (pathotypes) of Campylobacter jejuni strains in C57BL/6 interleukin-10 deficient mice, National Institutes of Health, Food and Water borne Diseases Integrated Research Network, Cambridge. MD, June 1-4, 2010.
- Parthasarathy G and Mansfield LS. 2009. Recombinant IL-4 (rIL-4) enhances Campylobacter jejuni invasion of Intestinal Pig Epithelial Cells (IPEC-1), Microbial Pathogenesis, 2009 Jul;47(1):38-6. Epub 2009 May 3, PMID: 19409975.
- Wilson DL, Rathinam VK, Qi W, Wick LM, Landgraf J, Bell JA, Plovanich-Jones A, Parrish J, Finley RL, Mansfield LS and Linz JE. 2010. Genetic diversity in Campylobacter jejuni is associated with differential colonization of broiler chickens and C57BL/6J IL-10 deficient mice. Microbiology, Papers in Press. doi:10.1099/mic.0.035717-0.
- Jerome, J.P., J.A. Bell, A.E. Plovanich-Jones, J.E. Barrick, C.T. Brown, L.S. Mansfield. 2010. Standing genetic variation in contingency genes drives the adaptation of Campylobacter jejuni to a novel host. PloS One, In press.
- Bell, J.A., J. R. Gettings, J. P. Jerome, J. J. Kopper, A. Plovanich-Jones, V. A. K. Rathinam, J. L. St. Charles, E.J. Smith, A. G. Staunton, and Mansfield, L.S. 2010. Campylobacter jejuni genome content and virulence phenotypes (pathotypes) in C57BL/6 IL-10 KO mice, submitted to Infection and Immunity.
|
Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Invited Speaker, National Institutes of Health, Food and Water borne Diseases Integrated Research Network, "Michigan State University Microbiology Research Unit Progress", 2009 Annual Meeting, Stevenson, WA. 3/29 - 4/1/2009. Gave the following talks: J.A. Bell, J.L. St. Charles, A.J. Murphy, V.A. Rathinam, A.E. Plovanich-Jones, E.L. Stanley, J.E. Wolf, J.R. Gettings, T.S. Whittam, Tristan Lefebure, Michael Stanhope, L.S. Mansfield. 2009. Pathogenicity of Campylobacter jejuni strains in C57BL/6 IL-10-/- mice varies with strain genetic background, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. J.P. Jerome, A.E. Plovanich-Jones, J.A. Bell, Linda S. Mansfield. 2009. Genetic changes during serial passage of Campylobacter jejuni in C57BL/6 IL-10-/- mice are associated with enhanced virulence, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. J.D. Olmstead, V.A.K. Rathinam, L.S. Mansfield. 2009. Determining organ specific recovery of Campylobacter jejuni in a murine disease model, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. JL St. Charles1,3, JA Bell1, and LS Mansfield. 2009. Development of murine models for study of Guillain-Barré Syndrome, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. Several students were mentored for study of enteric diseases of food animals. Five students are pursuing PhD degrees in food safety related to these enteric pathogens including John Paul Jerome, Jessica St. Charles, Jamie Jennifer Kopper, Ankit Malik and Vijay Rathinam. Three of these students have passed their preliminary exams. Two are preparing for this exam. One, Vijay Rathinam, successfully defended his Ph.D. thesis and obtained a postdoctoral position at the University of Massachusetts at Wooster, MA where he is studying the pathogenesis of inflammatory bowel disease due to bacterial pathogens. Two undergraduate students working in the lab on these projects were accepted into veterinary schools (Bianca Buffa, Michigan State University and Jenna Gettings, North Carolina State University). Jenna Gettings will also complete a M.Sc. degree in public health as part of her veterinary curriculum. Dr. Mansfield organized a summer seminars in Food and Waterborne Diseases for the faculty and students of Michigan State University. People attending have come from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. This included Marc Basson, M.D., Ph.D. from Wayne State University, Stephen Cendroski, Ph.D. from Michigan State University and Christopher Waters, Ph.D. from Princeton University. Also, Dr. Mansfield attended and presented at a scientific conference held by the National Institutes of Health at Skamania Lodge, Stevenson, Washington on Food and Water borne Pathogens. Dr. Mansfield will attend the USDA Multistate Research Project 2009 Meeting NC1041 in Chicago, Illinois on December 5th and 6th. PARTICIPANTS: Principal Investigator: Linda S. Mansfield (Station Representative), Julia A. Bell, Ph.D., Research Assistant Professor and the following graduate students worked on these projects; Graduate Students: Vijay A.K. Rathinam, B.V.Sc., Ph.D., John Paul Jerome, Ph.D. candidate, Jessica L. St. Charles, Ph.D. candidate, Jamie Jennifer Kopper, Ph.D. candidate, and Ankit Malik, Ph.D. candidate. Drs. Michael Stanhope at Cornell University, Dr. Michael Konkel at Washington State University, and Dr. Andrea Amalfitano at Michigan State University were collaborators. TARGET AUDIENCES: These studies and publications target scientists and medical and veterinary practitioners interested in food safety, bacterial pathogensis and immunology. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Multiple Factors Interact to Produce Responses Resembling Spectrum of Human Disease in Campylobacter jejuni Infected C57BL/6 IL-10-/- Mice Campylobacter jejuni infection produces a spectrum of clinical presentations in humans, including asymptomatic carriage, watery diarrhea, and bloody diarrhea, and has been epidemiologically associated with subsequent autoimmune neuropathies. This microorganism is genetically variable and possesses genetic mechanisms that may contribute to variability in nature. However, relationships between genetic variation in the pathogen and variation in disease manifestation in the host are not understood. We took a comparative experimental approach to explore differences among different C. jejuni strains and studied the effect of diet on disease manifestation in an interleukin-10 deficient mouse model. In the comparative study, C57BL/6 interleukin-10-/- mice were infected with seven genetically distinct C. jejuni strains. Four strains colonized the mice and caused disease; one colonized with no disease; two did not colonize. A DNA:DNA microarray comparison of the strain that colonized mice without disease to C. jejuni 11168 that caused disease revealed that putative virulence determinants, including loci encoding surface structures known to be involved in C. jejuni pathogenesis, differed from or were absent in the strain that did not cause disease. In the experimental study, the five colonizing strains were passaged four times in mice. For three strains, serial passage produced increased incidence and degree of pathology and decreased time to develop pathology; disease shifted from watery to bloody diarrhea. Mice kept on an ~6% fat diet or switched from an ~12% fat diet to an ~6% fat diet just before infection with an non-adapted strain also exhibited increased incidence and severity of disease and decreased time to develop disease, although the effects of diet were only statistically significant in one experiment. C. jejuni strain genetic background and adaptation of the strain to the host by serial passage contribute to differences in disease manifestations of C. jejuni infection in C57BL/6 IL-10-/- mice; differences in environmental factors such as diet may also affect disease manifestation. These results in mice reflect the spectrum of clinical presentations of C. jejuni gastroenteritis in humans and contribute to usefulness of the model in studying human disease. Study Impact - These findings provide cost effective animal models that can be used to study the genetic basis of virulence of the enteric pathogen C. jejuni. These murine models are expected to act as surrogates for humans to study C. jejuni strains coming from humans, cattle and swine. In 2008-2009, we used the models to establish the effect of host passage and diet on disease outcome (pathotype observed). We also screened a panel of C. jejuni strains from human patients in the murine model to observe whether variations in pathotype could be observed based on the genotype of the organism.
Publications
- Bell JA, St. Charles JL, Murphy AJ, Rathinam VA, Plovanich-Jones AE, Stanley EL, Wolf JE, Gettings JR, Whittam TS, Mansfield LS. 2009. Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10(-/-) mice, BMC Microbiology, 2009 Mar 18;9:57, PMID: 19296832.
- Rathinam VA, Appledorn DM, Hoag KA, Amalfitano A, Mansfield LS. 2009. Campylobacter jejuni-induced activation of dendritic cells involves cooperative signaling through TLR4-MyD88 and TLR4-TRIF axes, Infection and Immunity, 2009 Mar 30, PMID: 19332531.
- Rathinam VA, Hoag KA, and Mansfield LS. 2008. Dendritic cells from C57BL/6 mice undergo activation and induce Th1-effector cell responses against Campylobacter jejuni. Microbes and Infection 2008 Oct;10(12-13):1316-24. Epub 2008 Aug 5, PMID: 18725315.
- Mansfield LS, Patterson JS, Fierro BR, Murphy AJ, Rathinam VA, Kopper JJ, Barbu NI, Onifade TJ, and JA Bell. 2008. Genetic background of IL-10-/- mice alters host-pathogen interactions with Campylobacter jejuni and influences disease phenotype, Microbial Pathogenesis 2008 Oct;45(4):241-57 Epub2008 Jun 11.
- Parthasarathy G and Mansfield LS. 2008. Recombinant IL-4 (rIL-4) enhances Campylobacter jejuni invasion of Intestinal Pig Epithelial Cells (IPEC-1), Microbial Pathogenesis, Accepted April 22, 2009. ISSN 0882-4010.
- Abstracts V.A.K. Rathinam, Daniel M. Appledorn, Kathleen A. Hoag, Andrea Amalfitano and Linda S. Mansfield. 2009. TLR4→MyD88 and TLR4→TRIF signaling axes cooperate in activating dendritic cells in response to an intracellular enteric pathogen. Pattern Recognition Molecules and Immune Sensors, Keystone Meetings, Banff, Alberta CA, March 2009.
- J.A. Bell, J.L. St. Charles, A.J. Murphy, V.A. Rathinam, A.E. Plovanich-Jones, E.L. Stanley, J.E. Wolf, J.R. Gettings, T.S. Whittam, Tristan Lefebure, Michael Stanhope, L.S. Mansfield. 2009. Pathogenicity of Campylobacter jejuni strains in C57BL/6 IL-10-/- mice varies with strain genetic background, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- J.P. Jerome, A.E. Plovanich-Jones, J.A. Bell, Linda S. Mansfield. 2009. Genetic changes during serial passage of Campylobacter jejuni in C57BL/6 IL-10-/- mice are associated with enhanced virulence, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- J.D. Olmstead, V.A.K. Rathinam, L.S. Mansfield. 2009. Determining organ specific recovery of Campylobacter jejuni in a murine disease model, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
- JL St. Charles1,3, JA Bell1, and LS Mansfield. 2009. Development of murine models for study of Guillain-Barré Syndrome, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009.
|
Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: During this reporting period we have worked on several enteric pathogens that are of concern for food and water safety. We developed a tractable murine model for study of Campylobacter jejuni. This allows for determination of the genetic factors underlying disease caused by this pathogen. We have also screened Arcobacter isolates for their role in water borne outbreaks of enteric disease. We are on our way toward developing an animal model for this pathogen ro determine its ability to cause disease. I have mentored several students for study of enteric diseases of food animals. Four students are pursuing PhD degrees in food safety related to these enteric pathogens. Two of these have passed their preliminary exams. Two are preparing for this exam. One is getting ready to graduate. PARTICIPANTS: Our collaborators on these projects include Dr. Michael Stanhope at Cornell University and Dr. Michael Konkel at Washington State University. Vijay Rathinam BVSc, PhD candidate in the Comparative Medicine Integrative Biology Program performed some of this work in my laboratory for his PhD degree. He will graduate soon. Jenna Gettings and Dr. Julia Bell were my employees that worked on this project in my laboratory. TARGET AUDIENCES: I organized a seminar series in Food and Waterborne Diseases for the faculty and students of Michigan State University. People attending have come from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science and Human Nutrition departments. We invited several outside speakers including David Alland MD from New Jersey Medical School (Pathogenic E. coli), Shelly Rankin PhD from University of Pennsylvania (Pathogenic Salmonella), and Jimmy Ballard PhD from The University of Oklahoma (Clostridium difficile) I attended and presented at a scientific conference held by the National Institutes of Health at Callway Gardens Georgia on Food and Water borne pathogens. I am scheduled to attend the USDA Multistate Research Project 2008 Meeting NC1041 in Chicago Ilinois on December 6th and 7th. PROJECT MODIFICATIONS: There are no majors changes in the research project or procedures for this reporting period. We are following the specific aims we reported previously.
Impacts
We have published two papers, a book chapter and two abstracts this year describing the Campylobacter jejuni work. Both papers are in top grade journals. A paper is under preparation describing the findings of the Arcobacter work. We participated in assisting the Michigan Department of Community Health to identify the causative agent in a food borne outbreak in MI. Once basic CLIA laboratory work was completed to screen for bacterial pathogens, we assisted with innovative genetic testing of the isolates to further understanding of their pathogenic potential.
Publications
- Mansfield LS, Schauer DB, Fox JG. 2007. Animal Models of Campylobacter jejuni infections, In: Campylobacter, I. Nachamkin, C.M. Szymanski and M.J. Blaser (eds), American Society for Microbiology Press, Washington, D.C., Chapter 21:367-381.
- Mansfield LS, Patterson JS, Fierro BR, Murphy AJ, Rathinam VA, Kopper JJ, Barbu NI, Onifade TJ, and JA Bell. 2008. Genetic background of IL-10 knockout mice alters host-pathogen interactions with Campylobacter jejuni and influences disease phenotype, Microbial Pathogenesis 2008 Oct;45(4):241-57. Epub 2008 Jun 11.
- Rathinam VA, Hoag KA, and Mansfield LS. 2008. Dendritic cells from C57BL/6 mice undergo activation and induce Th1-effector cell responses against Campylobacter jejuni. Microbes and Infection 2008 Oct;10(12-13):1316-24. Epub 2008 Aug 5.
- Rathinam VA, Appledorn DM, Olmstead JD, Hoag KA, Amalfitano A and Mansfield LS. 2008. Campylobacter jejuni-induced Activation of Murine Dendritic Cells Involves Cooperative Signaling through MyD88 and TRIF. American Society for Microbiology, Boston, MA.
- Rathinam VA, Appledorn DM, Hoag KA, Amalfitano A and Mansfield LS. 2008. Campylobacter jejuni-induced Activation of Murine Dendritic Cells Involves Cooperative Signaling through MyD88 and TRIF. 15th International IRA Conference, Chantilly, VA.
|
|