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The 14th Annual Great Plains Infectious Disease Meeting We are pleased to host the 14th Great Plains Infectious Disease (GPID) Meeting which is returning to the University of Kansas after five strong years at the University of Missouri. The GPID meeting was originally developed to promote collaborations in the Great Plains region and to create a platform for networking among researchers. It continues to do so by promoting student, postdoc and young faculty’s research by hosting a poster presentation, and providing a platform for faculty, especially those new to the region, to give oral presentations that provide an outline of their research programs and promote new collaborative interactions. This year, we are proud to host more than 100 participants from across the region with nearly 30 posters being presented. We also welcome participants from the Kansas City Area Life Science Institute, the Animal Health Corridor and the USDA who are embarking on new partnerships with our academic partners. This meeting has always been successful due to the generosity of our academic sponsors and selected vendors. Thank you all for attending! The GPID Programming Committee 2015

ACKNOWLEDGEMENTS Generous academic and industrial sponsors have contributed greatly to the success of this meetings. Many thanks to all of our 2015 GPID supporters!

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14th Annual Great Plains Infectious Disease Meeting Program Schedule THE GOOD, THE BAD AND THE UGLY

November 6-7, 2015 University of Kansas-Lawrence

Friday, November 6, 2015 (Naismith Ballroom - Springhill Suites, 6th and New Hampshire, Downtown Lawrence, KS) 4:00pm 6:00pm Welcome and Reception 6:00pm 6:30pm Dinner

Plenary Session (Naismith Ballroom) 6:30pm 8:30pm Wayne Carter (KCALSI) “Life Science Ecosystem in Kansas City Region”

Blake Hawley (IAH) “Leveraging the Animal Health Corridor and Human Research to Build a Company” Dave Hustead (Boehringer Ingelheim) “It’s harder to decide which vaccines to give puppies and kittens than which vaccine to give kids”

Saturday, November 7, 2015 (Atrium, School of Pharmacy, West Campus ) 7:15am 7:55am Registration and Breakfast 8:00am 8:02am Opening Remarks

Session I - The Good: Immunology and Vaccine Development (2020 SOP) 8:02am 8:30am Jodi McGill (KSU) “The role of gamma delta T cells in immunity to respiratory infections in the

bovine”

8:30am 9:00am Brad Bearson (USDA) “DIVA Design: a cross-protective Salmonella vaccine” 9:00am 9:30am Tracy Nicholson (USDA) “Non-antibiotic Strategies to Prevent and Control Respiratory Bacterial” 9:30am 10:00am Mary Markiewicz (KUMC) “The influence of the innate immune receptor NKG2D on microbiota

composition and autoimmune diabetes"

10:00am 10:20am Refreshments and Networking Break (Atrium SOP) Session II - The Bad : Bad Bugs (2020 SOP) 10:30am

11:00am A. Paige Adams (KSU-Olathe) "Current Status of Vaccines for Venezuelan and Eastern Equine Encephalitis."

11:00am

11:30am Jeff Adamovicz (MU, LIDR) “New RB51 Vaccine Schedule for Cattle, Impacts on Immunity and Transmission Following B. abortus Infection.”

11:30am 12:00pm Eric Nicholson (USDA) “Thermodynamic insight into the basis of genetic prion disease in cattle” 12:00am 12:30pm Carl Gelhaus (MRIGlobal) “Animal Models for Tularemia Medical Countermeasures” 12:30 1:30pm Lunch - Mortar and Pestle Cafe (First Floor Dining Area)

Session III - The Ugly: Antibiotic Resistance (2020 SOP) 1:30pm 2:00pm Josie Chandler (KU) “Quorum sensing-controlled antibiotics and antibiotic-resistance factors and their

role in interspecies competition” 2:00pm 2:30pm Brian Brunelle (USDA) “Antibiotic exposure can induce various bacterial virulence phenotypes in

multidrug-resistant Salmonella enterica serovar Typhimurium” 2:30pm 3:00pm Mario Rivera (KU) “Targeting iron homeostasis for developing novel antimicrobials” 3:00pm 3:45pm Jim Riviere (KSU) “Use of Mixed-Effect and Physiological Based Pharmacokinetic Models of

Antimicrobial Drugs as a Framework for Interspecies Extrapolations”

Session IV - Poster Presentations 3:45pm 5:30pm Poster Session (Atrium SOP) 4:00pm 6:00pm Light Dinner / Hors d'oeuvres

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14th Annual Meeting

Plenary Speaker Session

Abstracts and

Bio-Summaries

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Wayne O. Carter, DVM, PhD, Dipl. ACVIM President and Chief Executive Officer Kansas City Area Life Sciences Institute Abstract: Life Science Ecosystem in Kansas City Region The greater Kansas City region spanning from Kansas State University in Manhattan, KS to the University of Missouri in Columbia, MO is home to a growing and vibrant life science ecosystem comprising over 250 companies and employing over 24,000 people. This region is also recognized as the Animal Health Corridor, home to the largest concentration of animal health R&D assets in the world with companies representing 75% of the global sales and over $19 billion. The region contains over 90 contract research organizations, is home of the $1.25 billion NBAF laboratory, and vibrant entrepreneurial and collaboration networks. We are growing the life sciences looking for nexus opportunities between human and animal diseases, health IT and convergent technologies intersecting with the life sciences.

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Wayne O. Carter, DVM, PhD, Dipl. ACVIM President and Chief Executive Officer Kansas City Area Life Sciences Institute Bio-Summary: Dr. Wayne Carter is President and Chief Executive Officer of Kansas City Area Life Sciences Institute that serves to advance human and animal health through translational research, collaboration and commercialization.

Dr. Carter is an experienced life science executive with both human and veterinary expertise spanning pharmaceutical and nutritional products. He is a technology aficionado, passionate about applying technology to improve healthcare decisions. Dr. Carter attended Purdue University, earning his BS degree in 1980, DVM in 1984 and practiced veterinary medicine for 5 years. He completed advanced residency training in internal medicine and board certification in 1992 and a PhD in cellular immunology at Purdue University in 1994.

Dr. Carter's passion for technology applications developed from a group started at Pfizer in 1997 focused on improving decisions in the drug development process. In that role, he was responsible for translational research and led the company’s North American clinical facilities in the development, validation and implementation of new clinical technologies to accelerate development decisions for human pharmaceuticals. He drove decisions at Pfizer by return on investment and grew the Clinical Technology group to span all phases of drug development and all therapeutic areas. In 2007, Dr. Carter was hired as Vice President of Research by Hill’s Pet Nutrition, a Colgate subsidiary, where he led nutrigenomics research and applied gene expression profiling to drive nutrition decisions for novel products in development.

Dr. Carter serves on several advisory boards. He serves on the board for the One Health Commission. He is Chairman of BioKansas, a non-profit organization to promote bioscience development in Kansas, a member of the Missouri Biotechnology Association Board of Directors, a non-profit organization to promote bioscience development in Missouri, a member of the KU Center for Research, Inc. Board of Trustees, a member of the University of Missouri Research and Discovery Advisory Board, member of the Kansas-State Olathe Advisory Board, a member of the MRIGlobal Board of Trustees, and a member of the Biological Sciences Advisory Board for the University of Kansas.

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Blake Hawley, DVM, MBA President and CEO North and South America Integrated Animal Health Kansas City Animal Health Corridor Abstract: Integrated Animal Health (IAH) is commercialising disruptive, patented technologies for the livestock sector and companion animal and horse space History Originally focused on a single technology platform, the IAH portfolio is expanding rapidly. The current focus is on commercialisation of its natural feed inclusion technologies which originated from licensing a human biotechnology firm’s asset. Upon visiting the Animal Health Corridor Homecoming event in 2013, Founder Rob Neely elected to set up an office in the US. Today, with headquarters in the BTBC and stemming from a single technology, IAH is now approaching 30 technologies in under two years, producing one of the most robust pipelines in privately held animal health companies.

Products IAH products range from natural feed inclusion technologies proven in feed trials over 12,000 dairy cows, to equine nutrition products and supplements, calf scour preventive and therapeutics, to poultry feed products, vaccines for livestock and pets, fly and tick repellents, cancer therapies, peptide products for weight loss and joint disease and much more. The vast majority originated from human health care and research.

Overall Objectives Leverage a lean, core team, their networks, and a strategic cohort of ‘early adopter’ commercial trial partners converted into active, paying customers. In parallel, expand globally by leveraging a solid advisory board, university supported technical white papers, evidence-based marketing materials, real-world testimonials and established market channels and partners. Our biggest opportunities lie in the global application and licensing of these technologies to enhance milk and meat production worldwide, which will then fund development of our broader pipeline.

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Current Commercial Products: Problem Solved and Market Demand

Animal health management poses one of the greatest challenges to the financial viability of producers and if not managed, is a threat to food safety, biosecurity and community health. By the year 2050, the world will need to double food production to feed a global population estimated to be 9.1 billion*. Meeting this need involves the cost-effective production of safe, high-quality animal protein.

As livestock production continues to increase in response to rising demand and increased standard of living, innovative animal health solutions are needed in greater volumes. There is growing importance of livestock supplements, medicines and vaccines as a component of global food supply. A key challenge for all players in the food ecosystem is how to meet the so-called “productivity imperative” – how to produce more with less.

By providing ways for farmers and livestock producers to cost-effectively increase productivity, Integrated Animal Health can help address this challenge. Vaccines and medicines, feed additives, improved production practices and advanced techniques – all of these assist producers with preventing diseases and optimizing the efficiency of the feed-to-meat conversion for protein and milk yield.

A major challenge for the industry, however, is the increasing scrutiny and regulation of antibiotics given to animals. The use of antibiotics in the feed of dairy and their use as growth promoters in livestock are a significant public health concern, with international pressure building to abate or ban antibiotic use in animal production. The EU was the first region to implement a ban in 2006. Developed countries such as the US, Canada and Australia are under increasing pressure to follow suit and major corporations such as McDonald’s are committing to antibiotic-free meats.

“…WHO has long recognized that antibiotic use in food animals contributes importantly to the public health problem of antibiotic resistance”. – World Health Organization, March 12, 2012

Widespread scientific concern about the extensive use of antibiotics in the food chain is creating opportunities for:

• Non-antibiotic methods of enhancing animal productivity and profitability • Increasing the innate immune function so disease doesn’t have so great an impact • Helping prevent infections by eliminating exposure to insect-borne vectors such as flies

and mosquitoes • Natural answers to improving food animal growth & feed conversion efficiency

IAH’s range of natural products solve multiple layers of these issues.

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Blake Hawley, DVM, MBA President and CEO North and South America Integrated Animal Health Kansas City Animal Health Corridor

Bio-summary:

Blake Hawley is the President and CEO for Integrated Animal Health, commercializing disruptive, patented, natural technologies, biologics and biopharmaceuticals for the livestock sector, companion animals and horses. Prior to joining IAH, he consulted for Jaguar Animal Health, and served as Chief Commercial Officer for Kindred Biosciences, a pet biopharmaceutical company he helped take public in 2014. Blake has also served as Worldwide Director of Global Digital for Hill's Pet Nutrition, a subsidiary of Colgate-Palmolive, working in various roles from 1998-2013 including Managing Director for Hill's UK and Ireland business, Regional General Manager for Russia and Central-Eastern Europe and General Manager-Australia and New Zealand. Before joining Hill's, he worked for Kaytee Products Inc, servicing the pet bird, exotics and wild bird markets. He was Executive Director of an environmental non-profit organization, and served on several boards. He was an adjunct professor at the University of Wisconsin veterinary School. Today, he serves on the MBA Advisory Board for KU, the Board of Directors for the Lawrence Humane Society and the Board of Economic Development Corporation of Lawrence and Douglas County.

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David R. Hustead, DVM MPH DACVPM Director Veterinary Medical Affairs Pet Division Boehringer Ingelheim Vetmedica Abstract: It’s harder to decide which vaccines to give puppies and kittens than which vaccine to give kids Guidelines for the use of vaccines in pets direct veterinarians to give a small group of vaccines to essentially every patient while at the same time directing them to give a much larger set of vaccines only to those patients that have adequate risk of exposure. This is in contrast to pediatricians who give most of the available vaccines to every child they see while conducting risk assessment on only a very small number of vaccines. In the real world of a busy veterinary practice, it is difficult to conduct a good risk assessment interview during routine history taking. To assist veterinarians in this task, we provide a web based interview tool which asks pet owners simple yes or no questions about pet behaviors considered important to confer risk of exposure. The tool has a simple decision system that can be configured by the veterinarian which informs the pet owner if there is adequate risk of exposure to justify implementing a preventive program including vaccination. The tool also provides the pet owner with disease information and advice actions they can take to prevent these diseases in their pets and in the case of zoonotic diseases to prevent these diseases in the human family, too.

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David R. Hustead, DVM MPH DACVPM Director Veterinary Medical Affairs Pet Division Boehringer Ingelheim Vetmedica. Bio-summary: Dr. Hustead is a graduate of Kansas State University. Following several years in companion animal and equine practice, he joined Fort Dodge Laboratories as their first technical service’s veterinarian specializing in companion animal and equine health issues. During his tenure at Fort Dodge, he was responsible for both in-house and field professional services, planned and supervised R&D product approvals, supervised regulatory affairs activities in the US and internationally. He has served on several committees including the American Association of Feline Practitioners Vaccination Guidelines, American Veterinary Medical Association’s Council on Biologic and Therapeutic Agents and represented US animal health manufacturers on the VICH Expert Working Group on Pharmacovigilance. He is currently Director, Veterinary Medical Affairs, Pet Division, at Boehringer Ingelheim Vetmedica. His team of veterinarians is responsible for technical consultations with customers, technical training of sales personnel, ensuring technical appropriateness of sales activities, post-approval clinical studies, interactions with organized veterinary medicine and liaison with colleges of veterinary medicine. Dr. Hustead recently obtained a Masters of Public Health from University of Iowa/Iowa State University and is a diplomat in American College of Veterinary Preventive Medicine.

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14th Annual Meeting

Oral Presentation

Abstracts

Session I

THE GOOD: Immunology and Vaccine

Development

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Jodi L. McGill, Ph.D. Assistant Professor Immunology Department of Diagnostic Medicine and Pathobiology Kansas State University Abstract: The role of gamma delta T cells in immunity to respiratory infections in the bovine γδ T cells are non-conventional T lymphocytes that form an important bridge between the innate and adaptive immune systems. They are conserved in all vertebrate species examined so far, but are particularly abundant in the immune system of the ruminant. Thus, cattle are an ideal model for studying the role of nonconventional T cells in immunity to infections and disease. We study the response of γδ T cells to infection with Mycobacterium bovis, the causative agent of bovine and zoonotic tuberculosis; and to bovine respiratory syncytial virus infection, a common respiratory viral infection of young calves and human infants. We have shown that γδ T cells secrete a number of inflammatory chemokines and cytokines in response to bacterial and viral infections both in vitro and in vivo, and play an important role in immune cell recruitment and inflammatory responses at the site of infection in the lungs.

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Jodi L. McGill, Ph.D. Assistant Professor Immunology Department of Diagnostic Medicine and Pathobiology Kansas State University Bio-summary: Jodi L. McGill received her B.S. in Microbiology from Iowa State University. She received her M.S. in Pathology and Ph.D. in Immunology from the University of Iowa studying dendritic cell biology and CD8 T cell immunity during influenza virus infection in a murine model. She did her post-doctoral fellowship at the National Animal Disease Center, USDA under the guidance of Dr. Randy E. Sacco studying the immune response to bovine respiratory syncytial virus and bovine tuberculosis. Dr. McGill started in her position as an Assistant Professor of Immunology in September 2014 in the Department of Diagnostic Medicine and Pathobiology at Kansas State University. The focus of her research is to elucidate the role of nonconventional T cells in shaping the innate and adaptive immune response to respiratory infections in humans and animals. Dr. McGill currently studies the response to several respiratory pathogens including bovine respiratory syncytial virus, Mannheimia haemolytica and Mycobacterium bovis, the causative agent of bovine and zoonotic tuberculosis.

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Brad Bearson, Ph.D. USDA, ARS Research Microbiologist National Laboratory for Agriculture and the Environment (NLAE) Ames, IA Abstract: DIVA Design: a cross-protective Salmonella vaccine Salmonella is a leading cause of bacterial foodborne disease and food-related death in the U.S. Greater than 50% of U.S. swine farms test positive for Salmonella, resulting in issues for both animal production and food safety. Vaccination of swine against Salmonella is a potential mitigation strategy to reduce or prevent pathogen colonization of livestock. However, there are over 2,400 Salmonella serovars and current vaccines may not provide adequate cross-protection against heterologous serovars. Furthermore, vaccination against Salmonella may interfere with surveillance programs that monitor the presence of Salmonella in swine herds. To overcome current vaccine limitations, we rationally designed, constructed, and evaluated a Salmonella vaccine to provide broad protection against a variety of Salmonella serovars while allowing the differentiation of infected from vaccinated animals (DIVA). The attenuated Salmonella DIVA vaccine is anticipated to protect animal health while enhancing food safety.

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Brad Bearson, Ph.D. USDA, ARS Research Microbiologist National Laboratory for Agriculture and the Environment (NLAE) Ames, IA Bio-summary: Brad Bearson is a Research Microbiologist at the USDA, ARS, National Laboratory for Agriculture and the Environment (NLAE) in Ames, IA. He received a B.S. in Biomedical Sciences from the University of South Alabama (USA) in Mobile, AL and earned his Ph.D. in Basic Medical Sciences with an emphasis in Microbiology from the USA College of Medicine. He performed an American Society for Microbiology, Clinical Microbiology Post-doctoral Fellowship at the University of California at Los Angeles (UCLA). Brad was a Visiting Research Scientist at the Poultry Research Unit at Mississippi State, MS prior to his move to Ames, IA where his initial position was as a Research Molecular Biologist at the National Animal Disease Center. In 2004, Brad assumed his current position at NLAE and investigates Salmonella colonization and pathogenesis of the swine and turkey gastrointestinal tracts for the development of interventions strategies to protect animal health, enhance food safety, reduce economic losses for producers, and limit environmental risk. In conjunction with his wife, Shawn Bearson, he has rationally designed, constructed and evaluated an attenuated Salmonella enterica serovar Typhimurium vaccine that allows the differentiation of infected from vaccinated animals (DIVA) while providing broad protection against multiple Salmonella serovars. To date the attenuated S. Typhimurium vaccine has been evaluated in both swine and turkeys for protection against S. Typhimurium and S. Choleraesuis or multidrug-resistant S. Heidelberg, respectively. A PCT International Application has been filed for the S. Typhimurium DIVA vaccine with a publication number of WO 2015/103104 A1.

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Tracy Nicholson, Ph.D. Research Microbiologist National Animal Disease Center U.S. Department of Agriculture Ames, IA Abstract: Non-antibiotic Strategies to Prevent and Control Respiratory Bacterial Infections in Swine The U.S. swine industry is the second largest producer of pork in the world, and respiratory disease in pigs is the most important health concern for swine producers today, making the development of efficacious vaccines and therapeutic interventions that can protect against respiratory infections a top research priority. The primary bacterial agents responsible for respiratory disease in swine include Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Actinobacillus pleuropneumoniae, and Pasteurella multocida. In addition to causing pneumonia, bacteria such as Haemophilus parasuis and Streptococcus suis and can cause systemic diseases such as meningitis, polyserositis, arthritis and septicemia. Antibiotics can be costly, of limited value against mixed infections, and may eventually be limited in their use in domestic livestock. In addition, several bacteria, such as Methicillin Resistant Staphylococcus aureus (MRSA), while causing minimal disease in swine, are zoonotic agents with potential economic consequences to the swine industry. The overall goals of my research are to: (1) identify the transmission, genetic, and pathogenic mechanisms used by these bacterial pathogens (2) develop and evaluate candidates for improved diagnostic tests, vaccines, biotherapuetic, and novel non-antibiotic strategies to prevent, reduce, or eliminate colonization and diseases caused by these pathogens.

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Tracy Nicholson, Ph.D. Research Microbiologist National Animal Disease Center U.S. Department of Agriculture Ames, IA Bio-summary: Tracy Nicholson is from San Antonio, Texas. Dr. Nicholson received a B.S. degree in Biomedical Science from Texas A&M University in 1995. She stayed at Texas A&M for graduate school where she worked in the laboratory of Dr. Andreas Bäumler studying Salmonella Typhimurium pathogenesis and earned her Ph.D. in Medical Microbiology in 2000. After graduation, she spent 4 years working as a NIH Postdoctoral Research Fellow with Dr. Richard Stephens at University of California, Berkeley, investigating global gene expression in the obligate intracellular bacterium Chlamydia trachomatis during active and persistent infections. In 2004, she began working for the USDA in Ames, Iowa, as a Principal investigator and team member for the Strategies to Control and Prevent Bacterial Infections in Swine research project. Her research encompasses identifying and assessing virulence mechanisms of bacterial pathogens associated with swine (Bordetella bronchiseptica, Haemophilus parasuis, Stretococcus suis, and methicillin-resistant Staphylococcus aureus), countermeasures to prevent infection, and host immune response to infection. She has authored/co-authored 34 peer-reviewed publications, member of the Editorial Board for the journals Pathogens and Infection and Immunity, and is currently serving as chair for Division Z Animal Health for the American Society for Microbiology.

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Mary Markiewicz, PhD Assistant Professor Department of Microbiology, Molecular Genetics & Immunology University of Kansas Medical Center Abstract: The influence of the Innate Immune Receptor NKG2D on Microbiota Composition and Autoimmune Diabetes Mary Markiewicz Department of Microbiology, Molecular Genetics & Immunology, University of Kansas Medical Center, Kansas City, KS USA Type 1 diabetes is an autoimmune disease caused by T cell-mediated destruction of the insulin-producing β cells in pancreatic islets. The current standard of care for type 1 diabetes patients is insulin replacement. While effective, this treatment is cumbersome and the life expectancy of type 1 diabetics is still reduced compared with non-diabetics. Therefore, novel prevention strategies and therapies are urgently needed. To reach this goal, a better understanding of the molecular mechanisms of β cell destruction is needed. Both human and animal studies implicate signaling through the NK group 2 member D (NKG2D) immune receptor in autoimmune diabetes pathogenesis. However, the mechanism by which NKG2D influences diabetes development is unclear. New data from our lab utilizing the non-obese diabetic (NOD) mouse model suggest that NKG2D interaction with its ligands affects β-islet cell destruction and diabetes via multiple and opposing mechanisms. First, NKG2D-ligand interactions promote a diabetes-protective intestinal microbiota composition. Second, NKG2D-ligand interaction via homotypic T cell contact during CD8+ T cell differentiation into cytotoxic T lymphocytes (CTLs) decreases diabetes-enhancing cytokine production. Third, NKG2D-ligand interaction via homotypic T cell contact within pancreatic islets enhances lytic granule release and β-islet cell killing. We are continuing studies with the NOD mouse model as well as human pancreatic samples to clearly define these roles of NKG2D receptor-ligand interaction in autoimmune diabetes pathogenesis.

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Mary Markiewicz, PhD Assistant Professor Department of Microbiology, Molecular Genetics & Immunology University of Kansas Medical Center Bio-summary: Mary Markiewicz received her Ph.D. in Immunology from the University of Chicago from her studies investigating the requirements for an effective tumor-specific CD8+ T cell response. She continued her tumor immunology studies during a one-year postdoctoral fellowship at the Cardinal Bernardin Cancer Center at Loyola University in Chicago in Dr. Martin Kast’s laboratory. She next moved to St. Louis for a postdoctoral fellowship in Dr. Andrey Shaw’s laboratory in the Department of Pathology and Immunology at Washington University School of Medicine where she began her studies concerning the role of the natural killer (NK) cell activating receptor NKG2D and its ligands in various types of immunity. She then stayed on at Washington University as an Instructor and Research Assistant Professor and focused her attention on determining the role of NKG2D and its ligands in autoimmune diabetes and tumor immunity. In 2014 she joined the faculty of the Department of Microbiology, Molecular Genetics & Immunology at the University of Kansas Medical Center, continuing her studies to understand the role of NKG2D signaling in autoimmunity and tumor immunity. She has received funding for her work from the American Cancer Society, the American Diabetes Association and the NIH.

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14th Annual Meeting

Oral Presentation Abstracts

Session II

THE BAD: Bad Bugs

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A. Paige Adams, DVM, PhD Research Assistant Professor Kansas State University Olathe Olathe, KS Abstract: Current Status of Vaccines for Venezuelan and Eastern Equine Encephalitis A. Paige Adams, DVM, PhD Kansas State University, Olathe, KS 66061 Venezuelan (VEEV) and eastern (EEEV) equine encephalitis viruses are closely related single-stranded, positive-sense RNA viruses and are members of the Alphavirus genus of the family Togaviridae. Both viruses are capable of causing encephalitis in animals and humans when transmitted by mosquito vector or potentially via infectious aerosol. Periodic re-emergence of Venezuelan equine encephalitis (VEE) in Central and South America and the yearly threat of eastern equine encephalitis (EEE) in North America emphasize the importance of these pathogens to public health and veterinary medicine. Despite their importance and over 60 years of research on VEE and EEE, there are no licensed human vaccines or effective antiviral treatments for human or equine disease, and the concern that VEEV and EEEV may be used for biological terrorism has also underscored the need for the development of improved, licensed vaccines and effective antiviral treatments. Recent advances in genetic engineering and vaccine design have been exploited to produce new vaccine candidates for VEE and EEE. This presentation will review: 1) past and present approaches for the development of these vaccines, 2) efficacy of vaccine candidates in various animal models, 3) limitations and advantages of each approach, and 4) the potential use of these vaccine candidates in the future.

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A. Paige Adams, DVM, PhD Research Assistant Professor Kansas State University Olathe Olathe, KS Bio-summary: Dr. Adams currently serves as Research Assistant Professor at K-State Olathe and is an ancillary faculty member of the Department of Diagnostic Medicine and Pathobiology at Kansas State University, College of Veterinary Medicine. Dr. Adams, a native Texan, received her Bachelor of Science and Doctor of Veterinary Medicine degrees from Texas A&M University. After graduating from veterinary school, she received large animal internship training at the Atlantic Veterinary College, University of Prince Edward Island, Canada, and large animal surgery residency training at the College of Veterinary Medicine, University of Minnesota. Dr. Adams received a Ph.D. in immunology from Cornell University with NIH support from institutional and individual National Research Service Awards (NRSA). In 2004, she worked as a postdoctoral fellow in the laboratory of Dr. Scott Weaver at the University of Texas Medical Branch, where she studied pathogenesis of and vaccine development for Venezuelan (VEEV) and eastern (EEEV) equine encephalitis viruses. In 2009, she joined the faculty at UTMB, where she continued her work with VEEV and EEEV, including some work with arthralgic alphavirus, chikungunya virus (CHIKV). In 2010, Dr. Adams received NIH K08 funding to study the genetic factors that contribute to differences in neurovirulence of subtype IE VEEVs. This award also concentrated on studies related to equine amplification of VEEV as a major determinant of epidemic potential. Additionally, Dr. Adams served as a co-investigator on a Western Regional Center of Excellence (WRCE) NIH grant to study quantitative mapping of CD8+ T cell responses of nonhuman primates (NHP) to vaccine strains of CHIKV. In 2013, Dr. Adams joined the faculty at K-State Olathe, where she has been involved in teaching and supporting the Veterinary Biomedical Science (VBS) Master of Science program. Dr. Adams is an active member of several professional societies, including American Veterinary Medical Association (AVMA), American Association of Veterinary Immunologists (AAVI), American Society for Virology (ASV), and American Society of Tropical Medicine and Hygiene (ASTMH), and she serves as a reviewer for several scientific journals.

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Jeffery Adamovicz, Ph.D. Associate Professor Director, Laboratory of Infectious Disease Research College of Veterinary Medicine Laboratory University of Missouri Abstract: New RB51 Vaccine Schedule for Cattle, Impacts on Immunity and Transmission Following B. abortus Infection Jeff Adamovicz, Ph.D. Department of Veterinary Pathobiology, Columbia, Missouri, United States The current recommended vaccination for brucellosis in at-risk cattle is a single calf hood dose of the live attenuated vaccine RB51. While this vaccine is successful in reducing infections in cattle it is not fully infective in preventing vertical transmission, abortions or the development of chronic infection. The hypothesis of this study is that RB51 booster vaccinations will provide either increased cell mediated or humoral immunity as compared to a single calf-hood vaccination alone. This research includes four specific aims; 1) that multiple doses of RB51 vaccine will induce a measurable increase in cell-mediated and humoral immunity and that this will correlate with a decreased incidence of infection and abortion at challenge; 2) that multiple doses of RB51 will not induce abortion when an adult booster vaccination is given during pregnancy; 3) that we will be able to correlate cell-mediated immunity data with a predicted outcome at challenge to establish an immune correlate for vaccination; 4) that RB51 titers to multiple doses of vaccine will correlate with previous studies in terms of safety of the vaccine. For this study Black Angus cattle were divided into four treatment groups; three treatment groups (n=7 each) and a control group (n=4). Each treatment group was assigned to be vaccinated with zero, one, two, or three doses of vaccine. All cattle were treated under normal ranching conditions and were vaccinated for diseases following the usual protocols for the Wyoming region. Cell-mediated immunity was measured through semi-quantitative PCR, cytokine suspension array, flow cytometry, and cell proliferation and killing assays. Humoral immunity was measured through an RB51 ELISA. Cell killing assays were used to determine the up-regulation in cytokine expression and CTL cell killing. An immortalized bovine macrophage cell line was used to perform cell-mediated immunity assays. Results will be presented for assays listed above. The results will reflect data from all three RB51 vaccine doses. Vaccinated cattle and control cattle were challenged with wild type B. abortus S2308 to determine the efficacy of multiple doses on bacterial burden, incidence of vertical transmission and abortion and those results will be discussed along with an alternate recommended vaccination schedule for at risk cattle. 28

Jeffery Adamovicz, Ph.D. Associate Professor Director, Laboratory of Infectious Disease Research College of Veterinary Medicine Laboratory University of Missouri Bio-summary: Jeff Adamovicz is from Des Moines, IA. He received his BA in Biotechnology from the University of Northern Iowa and Ph.D. in the Department of Microbiology at the Uniformed Services University of Health Sciences in Bethesda, MD. He did a post-doctoral fellowship in the Bacteriology Division, USAMRIID. Subsequently he worked on development of a new recombinant vaccine for plague which has just completed phase 3 clinical trials. After completion of his 24 year military career including service as a U.N. weapons inspector, and Chief of Bacteriology at USAMRIID Dr. Adamovicz became a senior medical product developer at USAMRIID helping to develop the next generation anthrax vaccine which is in clinical trials. He then worked for Midwest Research Institute for three years in central Asia as Chief Scientist, to improve public health infrastructure and develop biocontainment facilities and procedures. Following that he then worked for four years at the University of Wyoming as an Assistant Professor with a focus on brucellosis vaccine development and diagnostic testing as the State Veterinary serologist. He recently completed a study in cattle demonstrating improved vaccine efficacy by altering the recommended schedule for the current RB51 vaccine. Upon moving to the University of Missouri this past summer as an Associate Professor in the College of Veterinary Medicine he took over as the new Director of the Laboratory of Infectious Disease Research, a high containment biosafety laboratory facility. His research interests include common pathogenic mechanisms of facultative intracellular bacteria and the development of dendritic cell targeted vaccines.

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Eric Nicholson, Ph.D. Research Chemist National Animal Disease Center U.S. Department of Agriculture Ames, IA Abstract: Thermodynamic insight into the basis of genetic prion disease in cattle Prion diseases or transmissible spongiform encephalopathies (TSEs), are an unusual class of infectious disease. The causative agent of TSEs is a misfolded form of a host-encoded protein. In humans and cattle, prion disease can originate through infectious, spontaneous, and genetic processes. The most common genetic prion disease in humans is caused by the substitution of a lysine for a glutamic acid at position 200 (E200K) resulting in a heritable form of CJD. Rodent models of E200K CJD have proven interesting but have significant limitations including the need to use human-mouse chimeric protein constructs to reproduce a disease phenotype. The analogous amino acid substitution in cattle (E211K) was first identified in a case of BSE in the U.S. in 2006 and subsequently found in the only living offspring of that animal. This EK211 heterozygous animal has been used to develop a herd of EE211, EK211, and KK211 cattle for research on genetic TSEs. To date, our results do not faithfully replicate the observations from transgenic rodent models. These results led us to pursue additional studies of the folding and stability of E211K prion protein. Our studies implicate differences in the unfolded state of the wild-type and E211K protein in genetic prion disease in cattle.

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Eric Nicholson, Ph.D. Research Chemist National Animal Disease Center U.S. Department of Agriculture Ames, IA Bio-summary: Dr. Eric Nicholson grew up on the plains of western Kansas and received a B.S. degree in Biochemistry from Kansas State University in 1993. He then pursued his Ph.D. research in the lab of Dr. J. Martin Scholtz at Texas A&M University studying the thermodynamics of protein folding, completing his degree in 1999. After graduation he began post-doctoral research in the lab of Dr. Susan Marqusee at UC Berkeley as a Damon Runyon Cancer Research Fellow investigating the folding and stability of the prion protein. In 2004, he began working for the USDA in Ames, IA as a Principal Investigator on the Transmission, Differentiation, and Pathobiology of Transmissible Spongiform Encephalopathies research project. His research encompasses various aspects of animal prion disease, including genetics of disease resistance/susceptibility, biochemical methods of TSE strain differentiation, and folding/misfolding of the prion protein.

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Carl Gelhaus, PhD Principal Scientist and Director of Bacteriology MRIGlobal Kansas City, MO Abstract: F. tularensis is a highly infectious bacteria of increased interest due to its potential use as a biological/bioterror weapon. Tularemia, the disease caused by F. tularensis, can result in severe morbidity and mortality. As such, therapeutics and vaccines are needed. Due to the low natural incidence of tularemia, FDA licensure of tularemia therapeutics and vaccines will require well-developed animal models of disease under the FDA Animal Rule. We have developed well characterized animal models of tularemia which closely resemble human tularemia. We are actively testing therapeutics and vaccines and present a recent example.

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Carl Gelhaus, PhD Principal Scientist and Director of Bacteriology MRIGlobal Kansas City, MO Bio-summary: Dr. Gelhaus is Director of Biodefense Programs in the Medical Countermeasures of MRIGlobal, where he directs cutting edge research to protect the world from biological threats. Dr. Gelhaus has 17 years of experience in the field of Immunology, covering a diverse range of topics. Dr. Gelhaus has 9 years of experience in infectious disease, with emphasis on bioweapons and bioterror threats. In particular, Dr. Gelhaus is an internationally recognized Burkholderia and tularemia research and has presented his research around the globe. At the University of Colorado Health Sciences Center, Dr. Gelhaus demonstrated that CD8 T cells were not necessary for islet allograft tolerance in a mouse model of diabetes. At the United States Army Research Institute of Infectious Diseases (USAMRIID), Dr. Gelhaus performed research on the role of toll-like receptors (TLR) involved in the pathogenesis of diseases caused by gram-negative select agent bacteria. Namely, the roles of TLR1, 2, 4, 6, and 9 were investigated in Burkholderia mallei, Burkholderia pseudomallei, Francisella tularensis, and Yersinia pestis. This work was conducted both in vivo and in vitro, using mouse models, including bone marrow derived dendritic cell cultures. Dr. Gelhaus participated in research efforts involving the recombinant F1-V plague vaccine under development at USAMRIID. Dr. Gelhaus investigated the role of quorum sensing in Y. pestis pathogensis. At Battelle, Dr. Gelhaus was involved in the development of animal models as a Study Director for Good Laboratory Practices regulated studies. This effort included development of associated assay to support licensure of medical countermeasures through the US Food and Drug Administration Animal Rule. Dr. Gelhaus has investigated the efficacy of vaccines and drugs using these models, including doxycycline, and nitrogen-containing bisphosphonates. Dr. Gelhaus has 9 years’ experience with bacterial select agents under BSL-3 and ABSL-3 conditions, including Bacillus anthracis, B. mallei, B. pseudomallei, F. tularensis, and Y. pestis. Dr. Gelhaus has also provided scientific support under contract with a client company, including molecular biology database support and testing antibacterial properties of household items under EPA regulations.

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14th Annual Meeting

Oral Presentation

Abstracts

Session III

THE UGLY: Antibiotic Resistance

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Josephine R. Chandler, PhD Assistant Professor Department of Molecular Biosciences University of Kansas Lawrence, KS Abstract: Quorum sensing-controlled antibiotics and antibiotic-resistance factors and their role in interspecies competition Many Proteobacteria coordinate the production of virulence factors and other extracellular goods using a type of density-dependent cell-cell signaling called quorum sensing (QS). Extracellular products or ‘public goods’ are shared among the population and vulnerable to cheating by non-producing individuals (e.g. QS-defective variants). Cheaters do not incur any production costs and have a fitness advantage over cooperating members of the population, which can ultimately cause the population to collapse. One way to restrain the emergence of cheaters is by co-regulating “private goods” (goods that only benefit the producing individuals) along with the public goods, which creates a disadvantage to cheating. Some bacteria, such as the soil saprophyte Chromobacterium violaceum, use QS to regulate secreted antibiotics that are important for interspecies competition. Our group has been investigating the relationship between QS, cooperation, and interspecies competition using a combination of laboratory evolution and dual-species competition models. In addition to antibiotics, C. violaceum uses QS to regulate antibiotic resistance determinants. In a C. violaceum laboratory evolution model QS-deficient cheaters readily emerge and increase in the population. However, when antibiotics are added to the growth medium the cheaters are growth-restricted, because they are more susceptible to the antibiotics. This improves the competitiveness of the population, because it increases the frequency of antibiotic-producing individuals. Our results demonstrate a mechanism where antibiotics may play an important role in cheater restraint and the maintenance of QS. This may be important for survival in mixed microbial communities or in antibiotic-treated infections.

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Josephine R. Chandler, PhD Assistant Professor Department of Molecular Biosciences University of Kansas Lawrence, KS Bio-summary: Josephine Chandler grew up in Iowa City, IA and received her B.S. in Microbiology from the University of Iowa and her Ph.D. in Microbiology at the University of Minnesota. During her Ph.D., Dr. Chandler’s work focused on cell-cell signaling in the opportunistic pathogen Enterococcus faecalis. After her Ph.D., Dr. Chandler moved to Seattle, Washington and did a post-doctoral fellowship in the Department of Microbiology at the University of Washington under the mentorship of Dr. E. Peter Greenberg. As a postdoc Dr. Chandler was funded by an NIH NRSA to study cell-cell communication (quorum sensing) in the soil bacterium Burkholderia thailandensis and its close pathogenic relative, Burkholderia mallei. In 2012, she became a Research Assistant Professor at the University of Washington, and in 2013 she moved to the University of Kansas where she is currently an Assistant Professor in the Department of Molecular Biosciences. Her current research program uses several bacterial species including Burkholderia and Pseudomonas aeruginosa to understand how quorum sensing-control of antibiotics and other competition factors may promote survival in mixed microbial communities.

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Brian Brunelle, Ph.D. Research Microbiologist ARS, USDA, Research Microbiologist National Animal Disease Center Ames, IA Abstract: Antibiotic exposure can induce various bacterial virulence phenotypes in multidrug-resistant Salmonella enterica serovar Typhimurium Salmonella is one of the most prevalent bacterial foodborne diseases in the United States and causes an estimated 1 million human cases every year. Multidrug-resistant (MDR) Salmonella has emerged as a public health issue as it has been associated with increased morbidity in humans and mortality in livestock compared to antibiotic sensitive isolates. It is known that antibiotics can have unintended consequences on bacteria, and the goal of my research has been to characterize some of these collateral effects in MDR Salmonella Typhimurium. We have found that various antibiotics can enhance a variety of virulence phenotypes in MDR S. Typhimurium, such as inducing cellular invasion, increasing antibiotic resistance, and stimulating horizontal gene transfer. These factors may underlie some of the clinical observations associated with MDR Salmonella virulence.

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Brian Brunelle, Ph.D. Research Microbiologist ARS, USDA, Research Microbiologist National Animal Disease Center Ames, IA Bio-summary: Brian Brunelle was born and raised in western Massachusetts. He received his BS in Biochemistry from Rensselaer Polytechnic Institute in upstate New York. After that, he attended graduate school at the University of California in Berkeley and earned a PhD in Infectious Diseases studying the phylogenetics and evolution of Chlamydia trachomatis. He then did a post-doctoral research project at the National Animal Disease Center in Iowa where he studied genetic factors in cattle associated with resistance and susceptibility to bovine spongiform encephalopathy. Currently, he is a research microbiologist in a Food Safety group at the National Animal Disease Center studying how antibiotics impact virulence in multidrug-resistant Salmonella.

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Mario Rivera, Ph.D. Professor Department of Chemistry Courtesy Professor Department of Molecular Biosciences University of Kansas Abstract: Targeting iron homeostasis for developing novel antimicrobials Mario Rivera Department of Chemistry, University of Kansas Most antibiotics target a limited number of crucial biological processes, including DNA replication, protein translation, and cell wall biosynthesis. Thus, there is a need to discover and validate novel biological targets in bacteria. To address this issue, we have been exploring a new direction in the development of antimicrobials, which targets bacterial iron homeostasis. Bacterial iron metabolism offers a major vulnerability because: (i) iron is essential and must be obtained from the host, (ii) the concentration of free iron in the host is vanishingly low (≈ 10-18 M), (iii) to survive, bacteria have evolved mechanisms to “steal” iron from their host, which depend on well-regulated iron homeostasis, and (iv) once in the bacterial cell, iron can be toxic and must be stringently regulated. Hence, iron homeostasis, which is critical for infection, offers an excellent target for anti-infectives. To disrupt bacterial iron homeostasis we are targeting the protein bacterioferritin (BfrB), which is present in bacteria but not in humans. BfrB has a spherical and hollow structure where ≈3,500 iron atoms can be stored in the form of an Fe3+ mineral. BfrB functions by: (i) utilizing O2 or H2O2 to oxidize Fe2+ and store Fe3+ in its internal cavity, and (ii) accepting electrons from Bfd to reduce Fe3+ in the internal cavity and release Fe2+ to the cytosol. The talk will summarize our efforts at discovering small molecules, which by targeting the BfrB/Bfd interaction disrupt iron metabolism in Pseudomonas aeruginosa and render the opportunistic pathogen vulnerable.

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Mario Rivera, Ph.D. Professor Department of Chemistry Courtesy Professor Department of Molecular Biosciences University of Kansas Bio-summary: Dr. Mario Rivera is a Professor in the Department of Chemistry and a Courtesy Professor in the Department of Molecular Biosciences at the University of Kansas. He received his Ph.D. in Chemistry from the University of Arizona in 1991. The Rivera lab has worked on the development of new NMR methodology for the study of heme proteins in solution and has elucidated the structure, function and dynamics of a number or proteins involved in bacterial heme-iron acquisition and in bacterial iron metabolism. Their findings have contributed to shape current understanding of iron homeostasis in P. aeruginosa. A current focus of the Rivera lab is to capitalize from their relatively recent and unprecedented structural finding, which revealed the structure of P. aeruginosa bacterioferritin (BfrB) in complex with its cognate partner (Bfd). The interaction between BfrB and Bfd is necessary to regulate bacterial cytosolic iron concentrations by enabling the crucial equilibrium between free Fe2+ and Fe3+ sequestered in BfrB. P. aeruginosa mutants lacking BfrB or Bfd are growth impaired in iron-restricted media, significantly less virulent than the parent strain in C. elgans host and hyper-susceptible to antibiotics. Consequently, the Rivera lab, in collaboration with a multidisciplinary team with expertise in synthetic organic chemistry (Richard Bunce, Oklahoma State University), medicinal chemistry (Blake Peterson) and microbiology (Josephine Chandler and Bill Picking, University of Kansas), is working toward validating the BfrB-Bfd binding interaction as target for the development of novel antibiotics.

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Jim E Riviere, DVM, PhD The MacDonald Endowed Chair in Veterinary Medicine University Distinguished Professor Kansas Biosciences Eminent Scholar Director, Institute of Computational Comparative Medicine College of Veterinary Medicine Kansas State University, Manhattan, KS Abstract: Use of Mixed-Effect and Physiological Based Pharmacokinetic Models of Antimicrobial Drugs as a Framework for Interspecies Extrapolations Out of necessity, veterinarians must work with multiple species of animals to assure that administered drugs are both efficacious, safe and do not produce residues in edible tissues of food producing animals. Making extrapolations across species is a daily task. Two pharmacokinetic modeling techniques can be used to make such extrapolations; physiological-based (PBPK) and mixed-effect population pharmacokinetics (PopPK). Both allow incorporation of disease effects on distribution (protein binding), elimination or biotransformation based on published literature. In fact, these approaches could be considered meta-analysis tools that allow relevant data to be incorporated in appropriate models. Efficiency of these approaches were assessed using published and new experimental data from pharmacokinetic studies on penicillin-G, oxytetracycline and flunixin in cattle and swine. The strengths and weaknesses of these approaches will be discussed in the context of extrapolating antimicrobial drug disposition data across laboratory animal and veterinary species to humans.

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Jim E Riviere, DVM, PhD The MacDonald Endowed Chair in Veterinary Medicine University Distinguished Professor Kansas Biosciences Eminent Scholar Director, Institute of Computational Comparative Medicine College of Veterinary Medicine Kansas State University, Manhattan, KS Bio-summary: Dr. Jim E. Riviere is the MacDonald Endowed Chair, Kansas Bioscience Eminent Scholar and a University Distinguished Professor at Kansas State University in Manhattan where he is the founding director of the Institute of Computational Comparative Medicine. He spent the first three decades of his career at North Carolina State University in Raleigh as a Burroughs Wellcome Fund Distinguished Professor in Pharmacology. He received his BS (summa cum laude) and MS degrees from Boston College, his DVM and PhD in pharmacology as well as a DSc (hon) from Purdue University. He is an elected member of the National Academy of Medicine, and serves on the National Research Council (NRC) Board of Agriculture and Natural Resources. He previously served on the Board of Scientific Counselors of the NIEHS National Toxicology Program, the FDA Science Advisory Board as well as on numerous NIH Study Sections, FDA Committees and journal editorial boards. He is the Editor of the Journal of Veterinary Pharmacology and Therapeutics and confounder of the USDA-supported Food Animal Residue Avoidance and Depletion (FARAD) program. His honors include the 1999 O. Max Gardner Award from the Consolidated University of North Carolina, the 1991 Ebert Prize from the American Pharmaceutical Association, the Harvey W. Wiley Medal and FDA Commissioner’s Special Citation, and the Lifetime Achievement Awards from both the European and American Association of Veterinary Pharmacology. Dr. Riviere has been PI on over 20 million dollars of extramural grants and published 550 full-length research papers and chapters, holds 6 U.S. Patents, and has authored/edited 11 books in pharmacokinetics, toxicology and food safety. His current research interests relate to the development of animal models; applying biomathematics to problems in toxicology, including the risk assessment of chemical mixtures, pharmacokinetics, nanomaterials, absorption of drugs and chemicals across skin; and the food safety and pharmacokinetics of tissue residues in food producing animals. Jim has been married to fellow toxicologist Dr. Nancy Monteiro-Riviere for 39 years and have three adult children.

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14th Annual Meeting

Poster Abstracts

Session IV

Poster Presentations

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14th Annual Great Plains Infectious Disease Meeting University of Kansas - Lawrence Kansas

November 6 - 7, 2015

SESSION IV: POSTER PRESENTATIONS

Poster #

Last Name

First Name

Abstract Title

1 Arizmendi Olivia IpaD from the T3SS of Shigella Triggers Macrophage Apoptosis

2 Behar Amanda Genetic Manipulation of Chlamydia trachomatis Inclusion Membrane Protein CT228 using the Adapted TargeTron System

3 Cherla Rama Coxiella burnetii infection inhibits Neutrophil Apoptosis through activation of Erk1/2 and Anti-apoptotic protein Mcl 1

4 Coate Eric Identification and validation of antigens associated with lethal pneumonic plague

5 Dhariwala Miqdad Bacterial activation of TLR7 in macrophages stimulates a non-canonical pathway for expression of type I interferon

6 Dowdell Alexander Localization of the B. burgdorferi Lipoproteome 7 Eleshy Rawan Antibiotic Resistance in Staphylococcus aureus Isolated From Cystic

Fibrosis Patients 8 Fairlamb Max The Cra-FruK complex alters regulation of central metabolism of in

γ-Proteobacteria 9 Garcia Brandon Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking

Activation of the C1 Complement Complex 10 Hermanas Timothy A c-di-GMP Signaling System in Bacillus anthracis Spores

11 Johnson Lauren Exploration of the Inhibitory Effects of Allicin on the Growth of Staphylococcus aureus in an In Vivo Animal Model

12 Klaus Jennifer Regulation of an Antibiotic-Induced Virulence Gene Cluster in Burkholderia pseudomallei

13 Krute Christina The Fatty Acid Kinase of Staphylococcus aureus Controls Virulence

14 Ledbetter Lindsey Mechanism of Vaccine-Induced Protection against Coxiella burnetii

15 Liang Lingfei Small-Angle X-ray Scattering Analysis of Bacteriophage Sf6 Structural Protein gp8: a Molecular Hub that Mediates the Assembly of the Tail Machine

16 Machen Alexandra Novel and Rapid Development of Stabilizers Against Infectious Disease Protein Toxins and Human Disease

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17 Matocha Kimberly NKG2D Genotype Alters Gut Microbiome Composition and Immunoglobulin A Production in The Large Intestine

18 McAllister Rachel The Induction of Small Colony Variant Staphylococcus aureus in Artificial Sputa Media

19 O’Neil Pierce Probing the Kinetic Stability of Tetanus Neurotoxin Using Biolayer Interferometry along with a Novel Denaturant Pulse-Chaperonin Assay with Parallel Confirmation Via Transmission Electron Microscopy

20 Olivarez Nicholas Bioinformatic Analysis of Antibiotic Resistance Mechanisms of Coxiella burnetii

21 Olson Rachel Amino Acids T45 and R53 in Yersinia pestis YopK Putative Binding Region Suppress Host Inflammasome Response

22 Park So Lee Thermostability of alphaviruses can be virus-specific

23 Pressnall Melissa Activation of Dendritic Cells by the subunit vaccine for Shigella

24 Sharma Neekun Constitutive Expression of the NKG2D Ligand RAE1ε Within Pancreatic Islets Ameliorates Autoimmune Diabetes Development in Non-Obese Diabetic Mice

25 Trembath Andrew Protective Roles for NKG2D in NOD Diabetes Development

26 Woehl Jordan Toward Understanding the Structural Basis of Inhibition of the Classical and Lectin Pathways of Complement by S. aureus Extracellular Adherence Protein

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Abstract #1

IpaD from the T3SS of Shigella Triggers Macrophage Apoptosis

Olivia Arizmendi1, William D. Picking2 and Wendy L. Picking2

1Dept. of Molecular Biosciences, University of Kansas, Lawrence, KS, USA; 2Dept. of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA

Abstract:

Shigellosis, a type of bacillary dysentery, is an infectious gastrointestinal disease caused by Shigella spp. Approximately 165 million cases of shigellosis occur every year around the world, the vast majority of them in developing countries. High levels of antibiotic resistance, an increase in multidrug-resistant Shigella isolates and the lack of a licensed vaccine are factors that situate shigellosis as a public health problem, especially among young children. Shigella is able to cause death of resident macrophages in the gut to avoid bacterial clearance early after infection. Shigella is then able to colonize the intestinal epithelium and induce inflammation, which ultimately gives rise to the symptoms of dysentery and bacterial shedding. The virulence of Shigella is intimately tied to its Type III Secretion System (T3SS) for which invasion plasmid antigen D (IpaD) is a structural element. IpaD was recently shown to induce apoptosis in B lymphocytes in conjunction with an additional unknown factor. Furthermore, previous studies have established that IpaD is secreted at levels beyond what is needed for its role as the T3SS needle tip protein. Here, we present data substantiating our hypothesis that IpaD secreted by Shigella triggers an apoptotic pathway in macrophages as an early step in infection. To test this, we have applied a multidisciplinary approach that includes cell biology, immunology and molecular pathogenesis. Macrophages exposed to purified IpaD show clear morphological changes consistent with classical apoptosis such as membrane blebbing and bundles of condensed chromatin. Furthermore, we observed cytotoxicity as measured by LDH release and activation of specific caspases. A multiplex assay also provided a clear understanding of the cytokine release profiles caused by IpaD. We then tested our findings in Shigella infection of cultured macrophages, which confirmed this role for IpaD in Shigella pathogenesis. A required IpaD domain as well as possible binding partners were also identified. These findings allow us to conclude IpaD is a contributing factor to macrophage cell death during Shigella infection. Learning more about the ways T3SSs interact with host cells provides potential new targets for antimicrobial drug design.

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Abstract #2

Genetic Manipulation of Chlamydia trachomatis Inclusion Membrane Protein CT228 using the Adapted TargeTron System

Amanda Behar1, Cayla Johnson2, Derek Fisher2 and Erika Lutter1 1 Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA; 2 Department of Microbiology, Southern Illinois University Carbondale, Carbondale, IL, USA

Abstract:

Chlamydia trachomatis is the most frequently reported bacterial sexually transmitted

infection. Even after a C. trachomatis infection is treated, there is an increased risk for

the development of pelvic inflammatory disease and cervical cancer, but the mechanisms

are poorly understood. As an obligate intracellular pathogen, C. trachomatis usurps

many host cell-signaling pathways from within a membrane bound vacuole, called an

inclusion. C. trachomatis is also known to synthesize and secrete via the type III

secretion system, inclusion membrane proteins (Incs) that insert into the inclusion

membrane and serve as the interface between Chlamydia and the host. C. trachomatis is

the first bacterial pathogen observed to recruit myosin phosphatase (MYPT1) for means

of host cell exit, and does so through the chlamydial Inc protein, CT228. In this study,

the chlamydial TargeTron system was used to genetically inactivate CT228 in the C.

trachomatis genome. TargeTron insertion was confirmed by PCR and expression of the

CT229-CT224 operon of the mutant was verified by RT-PCR to rule out polar effects.

The CT228 mutant was verified to be deficient in CT228 production and MYPT1

recruitment by immunofluorescence. This study demonstrates successful gene

inactivation of the chlamydial protein CT228 and confirms the role of CT228 in MYPT1

recruitment. Additionally, these studies provide a platform to further investigate the role

of CT228 in chlamydial pathogenesis.

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Abstract #3

Coxiella burnetii infection inhibits Neutrophil Apoptosis through activation of Erk1/2 and Anti-apoptotic protein Mcl 1

Rama Cherla*, Laura Schoenlaub, Yan Zhang, and Guoquan Zhang

Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, Columbia, Missouri, USA

Abstract:

Coxiella burnetii is an obligate intracellular pathogen that causes acute and chronic Q

fever in humans. It is mainly transmitted by inhalation of contaminated dust from

infected animals such as cattle and sheep as well as in petting zoos. Understanding the

host defense mechanisms will provide critical information for discovery of novel

therapeutic targets against C. burnetii infection. Neutrophils are the first responders to

migrate into the infection site during bacterial pathogen invasion. Previous studies from

our group showed that both virulent C. burnetii Nine Mile phase I (NMI) and avirulent

Nine Mile phase II (NMII) bacteria can infect mouse bone marrow-derived neutrophils

and NMI infection induced more severe disease in neutrophil-depleted mouse, suggesting

neutrophils play an important role in host defense against C. burnetii primary infection in

mice. However, the mechanisms of interaction between C. burnetii and neutrophils

remain unknown. In this study, we examined if there is a difference between NMI and

NMII in modulating host cell apoptotic signaling in mouse bone marrow-derived

neutrophils by MTS assay and TUNEL staining. Interestingly, both NMI and NMII

bacteria were able to inhibit neutrophil preprogramed apoptosis. Immunoblotting analysis

indicates that NMII infection induces a decreased caspase 3 cleavage and increased

phosphorylated p38 MAPK and Erk1/2 kinases. Inhibitors of p38 (SB203508) and

Erk1/2(PD98059) kinases were able to reduce NMII infection induced anti-apoptotic

activity. In addition, activation of Mcl1, a pro-survival Bcl2 family member was also

detected in NMII-infected neutrophils at different time points post-infection.

Collectively, these data suggest that C. burnetii infection induced anti-apoptotic activity

depends on activation of MAP kinase and Mcl1 signaling pathway.

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Abstract #4

Identification and Validation of Antigens Associated with Lethal Pneumonic Plague Eric A. Coate1 and Deborah M. Anderson1

1Department of Veterinary Pathobiology, University of Missouri, Columbia, MO

Abstract:

Inhalation of Yersinia pestis causes primary pneumonic plague, a rapidly progressing

disease that becomes untreatable shortly after symptom onset. We have recently shown

that lethal disease correlates with the development of cardiac arrhythmia in a Brown

Norway rat pneumonic plague model. This profound response is accompanied by

elevated IL-6, IL-1β, TNF-α and decreased platelets, consistent with a cytokine storm

severe sepsis enabled by bacterial growth in the lungs, and further suggesting that

arrhythmias could be caused by specific Y. pestis antigens. IL-6 and IL-1β are commonly

found during bacterial sepsis caused by toll-like receptor 2- or 4-dependent responses to

the bacterial cell wall or outer membrane lipopolysaccharide (LPS). However, host TLR-

2 and TLR-4 do not recognize Y. pestis and the antigens inducing this systemic

inflammatory response during infection are unknown. In this work, we sought to better

understand the mechanism underlying these arrhythmias and to explore the potential to

use this information to develop methods for early detection of primary pneumonic

plague. Towards these goals, we have compared electrocardiograms from rats infected

with WT or mutant Y. pestis strains in order to identify arrhythmias that were dependent

on specific Y. pestis virulence factors. Our data suggest that ST interval changes are

associated with the virulence factor YopJ and requires secretion, but not bacterial

transport, of the iron-binding siderophore Yersiniabactin. Furthermore, neither

bacteremia, fever nor serum cytokines were consistently associated with ST interval

changes. Together the data provide new insight into understanding the severity of

pneumonic plague, and support further investigation of the ECG as a diagnostic tool for

plague, and perhaps other infectious diseases.

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Abstract #5 Bacterial Activation of TLR7 in Macrophages Stimulates a Non-Canonical Pathway for Expression of Type I Interferon

Miqdad O Dhariwala1 and Deborah M Anderson1

1Department of Veterinary Pathobiology, University of Missouri-Columbia, MO USA Abstract: Phagocytosis induces localization of Toll-like receptors (TLR) to endosomes where their

subsequent activation by nucleic acids triggers the expression of type I interferon (IFN).

TLR7 is known to localize to acidified- lysosome associated membrane protein 1

(LAMP1) positive vacuoles in the phagosomal pathway of dendritic cells where it

delivers an activation signal through the adaptor protein MyD88 followed by activation of

one or more interferon regulatory factors (IRFs) and NFκB. While this is viewed as a

general response to RNA released during phagocytosis, TLR7 is in many cases not the

main pattern recognition receptor (PRR) involved in initiating expression of type I IFN

and the issues underlying why TLR7 may only sometimes be used are not clear. In this

work, we show that TLR7 signaling in macrophages following phagocytosis of Yersinia

pestis leads to the MyD88-independent expression of IFNβ. Inhibition or absence of IRF3

or NFκB activation prevented IFNβ expression suggesting both transcription factors may

be activated through TLR7. Furthermore, we show that TLR7 localization to vacuoles

occurs following phagocytosis of Y. pestis, Escherichia coli or Staphylococcus aureus but

is evaded by the obligate intracellular pathogens Francisella tularensis and Brucella

abortus in a manner that is independent of phagosomal escape suggesting early events in

the phagocytic pathway may be involved in specifying the localization of TLR7. Together

these data identify a non-canonical TLR7 signaling pathway in macrophages activated by

the immune-evasive pathogen Y. pestis. Whether this pathway is affected by crosstalk due

to activation of other PRRs by the potent immune stimulators E. coli and S. aureus

remains unknown.

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Abstract #6 Localization of the B. burgdorferi Lipoproteome

Alexander S. Dowdell1, Shiyong Chen1, Christina Azodi1, Max Murphy1, Wolfram R. Zückert1 1Department of Microbiology, Molecular Genetics & Immunology; University of Kansas Medical Center; Kansas City, KS, USA

Abstract:

The spirochete bacterium Borrelia burgdorferi is the most common vector-borne

pathogen in the United States and a worldwide health risk. B. burgdorferi exhibits a

dualistic lifestyle, continually transitioning between arthropod vectors and vertebrate

hosts. In order to cope with such radical environmental changes, B. burgdorferi employs

many lipid-modified membrane proteins (lipoproteins) to serve in diverse roles ranging

from nutrient acquisition to immune evasion. The genes for these lipoproteins are

distributed across a highly fragmented genome, which is composed of a linear

chromosome and several circular and linear accessory plasmids. Until recently, research

has been primarily focused on a small subset of surface lipoproteins known to be

immunogenic that could serve as potential vaccine candidates. The majority of the

bacterium's lipoproteome has however been neglected, despite its crucial role to the

survival of the organism. Here, we used a tagged expression library and well-estabilished

techniques to localize each protein of the B. burgdorferi lipoproteome to a particular

cellular location. We demonstrate that the majority of lipoproteins are surface-exposed

and that the nonessential "accessory" plasmids are enriched in these surface lipoprotein

genes. Finally, we used in silico techniques to show conservation in multiple "families"

of lipoproteins and to explore the role of lipoproteins lacking experimental investigation.

Our work confirms previous paradigms of B. burgdorferi genome organization and lays

the groundwork for the future investigation of previously uncharacterized lipoproteins.

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Abstract #7 Antibiotic Resistance in Staphylococcus aureus Isolated From Cystic Fibrosis Patients Rawan G. Eleshy1 and Erika Lutter1 1Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA Abstract: Cystic fibrosis (CF) is one of the most common autosomal recessive genetic diseases

caused by a mutation in the CFTR gene. Mutations within this gene disrupt the function

of the ion channels and inhibit the transport of chloride ions across the epithelial

membranes. This leads to dehydrated thick mucus and impaired mucocillary clearance.

As a result, the environment within the CF lung airways becomes ideal for bacterial

colonization by several bacterial species. Staphylococcus aureus is one of the first

pathogens isolated from the CF lung and is prevalent throughout the life of CF patients.

It is very adept at developing resistance to antibiotics and as such is of great concern to

the medical community. This study investigates the prevalence of antibiotic resistance

and presence of antibiotic resistance genes within a subset of S. aureus isolates recovered

from CF patients of various ages: adults (over 18), adolescents (13-18), and children

(under 13). Prior studies have identified nine genes that are correlated with antibiotic

resistance in CF patients. Isolates were tested for the presence of any of these resistance

genes by PCR. Some isolates expressed up to four resistance genes, while others did not

express any of them. Therefore, susceptibility tests were performed to determine if these

isolates show a resistance phenotype. Surprisingly, some of the isolates that contained the

resistance genes did not show resistance in the susceptibility tests. In addition, some

isolates showed resistance on Kirby-bauer plates without expressing the genes.

Understanding the antibiotic resistance mechanisms of Staph aureus isolates from CF

patients will provide significant insights into the complexity of CF infections and may

help in future patient treatments.

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Abstract #8

The Cra-FruK Complex Alters Regulation of Central Metabolism in γ-Proteobacteria Max Fairlamb1, Dipika Singh1, Chamitha Weeramange1, Benjamin Rau1, Sarah Meinhardt1, Sudheer Tungtur1, Dr. Aron Fenton1, Dr. Liskin Swint-Kruse1 1Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA Abstract: The Gram negative γ–proteobacteria include E.coli, Shigella, plague-causing Yersinia, and Vibrio cholarae. In these organisms, one protein that is required for pathogenicity is the catabolite repressor activator (“Cra”). Cra deletion is not lethal, but the bacteria are no longer robust enough to become pathogenic. This is probably related to its role as a central regulator of carbon metabolism: Cra directly regulates more than 100 genes, including those of glycolysis, gluconeogenesis, and the Kreb’s cycle. In regulating these pathways, Cra enables the bacteria to switch between aerobic and anaerobic metabolism. Cra is a LacI/GalR homolog and has a function analogous to LacI: Cra binds to operator DNA sequences to regulate transcription of downstream genes; and DNA binding is diminished when Cra binds fructose metabolites. Of these, fructose-1-phosphate (“F1P”) is a strong inducer at micromolar concentrations. F1P is created when fructose is transported into the cell via a phosphotransferase system. In addition, fructose-1,6-bisphosphate (“F16BP”, a central metabolite in glycolysis) has been reported to be a weak (millimolar) inducer in E. coli, although not in Yersinia. Further, if F16BP acts as a simple inducer like F1P, it is difficult to rationalize the switching mechanism between aerobic and anaerobic metabolism. We serendipitously discovered a new, tight protein-protein interaction between Cra and fructose-1-kinase (“FruK”). This kinase interconverts F1P and F16BP and is itself regulated by Cra. Thus, we hypothesized that the interaction must have a critical role in regulating central carbon metabolism. Experiments with purified proteins show that the Cra-FruK complex enhances DNA binding relative to Cra alone. Further, whereas F16BP acts as an inducer for the Cra-DNA complex, it acts as an anti-inducer for Cra-FruK-DNA complex. In contrast, F1P induces both Cra-DNA and Cra-FruK-DNA. Thus, the Cra-FruK interaction (+/- F16BP) provides the missing event for switching E. coli between aerobic and anaerobic metabolism.

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Abstract #9

Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex Brandon L. Garcia1,3, Hui Zhi2, Beau Wager2, Magnus Höök2,3, and Jon T. Skare2

1Department of Biochemistry & Molecular Biophysics, Kansas State University, Manhattan, KS, USA; 2Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, Bryan, TX, USA; 3Center for Infectious and Inflammatory Diseases, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, USA Abstract:

Microbial pathogens have evolved a sophisticated arsenal of secreted and membrane associated proteins capable of recognizing, exploiting and/or inactivating host immune responses. Pathogens that traffic in blood, lymphatics, or interstitial fluids must adopt strategies to evade human innate immune defenses, notably the human complement system. Through recruitment of host regulators of complement to their own surface many human pathogens are able to escape complement-mediated attack. The etiologic agent of Lyme disease, Borrelia burgdorferi, expresses a number of surface exposed proteins that bind directly to factor H related molecules, which function as the dominant negative regulator of the alternative pathway of complement. Relatively less is known about how B. burgdorferi evades initiation of the classical pathway of complement despite the observation that some sensu lato B. garinii isolates are sensitive to classical complement. Furthermore, the classical pathway is involved in the development of the adaptive immune response that is quelled following borrelial infection. Here we report that the borrelial lipoprotein BBK32 is capable of potent and specific inhibition of the classical pathway by binding with high affinity to the initiating complex, the first component of complement (C1). Using a biochemical and biophysical approach we localized the anti-complement activity of BBK32 to its globular C-terminal domain. Mechanistic studies reveal that BBK32 acts by entrapping C1 in its zymogen form by binding and inhibiting the C1 subcomponent, C1r, which serves as the initiating serine protease of the classical pathway. To our knowledge this is the first report of a spirochetal protein acting as a direct inhibitor of the classical pathway and is the only example of a biomolecule capable of specifically and noncovalently inhibiting C1/C1r. By identifying a unique mode of complement evasion this study greatly enhances our understanding of how pathogens subvert and potentially manipulate host innate immune systems.

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Abstract #10 A c-di-GMP Signaling System in Bacillus anthracis Spores Timothy M. Hermanas1; Chung-Ho Lin2, and George C. Stewart1

1Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 2Center for Agroforestry, University of Missouri, Columbia, MO Abstract: Bacillus anthracis is a gram-positive spore-forming organism that causes fatal disease in

humans, cattle, sheep and goats. B. anthracis spore stability, combined with ability to

inflict lethal disease, has resulted in its use as a biowarfare agent. The 2001 Amerithrax

attacks, utilizing spores concealed in letters, resulted in five deaths and over $320 million

dollars spent decontaminating areas of release. The B. anthracis spore structure and

physiology, especially involving the outermost exosporium layer, are incompletely

understood. The exosporium plays a role in spore uptake and intracellular survival in the

host. Utilizing transposon mutagenesis, we identified a gene in the Sterne strain, bas3594,

as important for the proper assembly of the exosporium. The bas3594 mutant spores

exhibited decreased BclA deposition and microarray data revealed an expression profile

similar to that of bclA, the gene encoding the major surface glycoprotein of the spore.

BAS3594 is a GGDEF/EAL-domain-containing protein. GGDEF/EAL domains are

typically found in proteins that regulate intracellular levels of the messenger c-di-GMP,

which modulates gene function and host immune responses. The B. anthracis genome

also encodes nine other proteins that contain GGDEF/EAL domains. Expression of

mCherry-GGDEF/EAL-protein fusions revealed seven of these putative cyclic di-GMP

regulatory proteins are expressed during specific phases of B. anthracis growth and

sporulation. A subset of these proteins is contained within mature spores and incubation

of B. anthracis spores with GTP resulted in the generation of c-di-GMP showing

evidence diguanylate cyclase activity in mature spores.

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Abstract #11 Exploration of the Inhibitory Effects of Allicin on the Growth of Staphylococcus aureus in an In Vivo Animal Model

Lauren J. Johnson1, Fawn Beckman1, J. David McDonald Ph.D1 1Department of Biological Sciences, Wichita State University, Wichita, KS USA

Abstract:

The antimicrobial properties of Allicin, while long known, require further investigation in order

to evaluate this garlic-derived chemical as an anti-infective agent against Staphylococcus aureus

wound infection. S. aureus is well adapted to live on skin as either normal flora or as a pathogen.

Indeed, it is carried as normal flora by approximately one-third of all people. Thus, there is an

important ongoing clinical problem with wound infection by this pathogen along with the fact

that antibiotics continue to lose their effectiveness against strains. This has motivated us to

explore alternative methods to deal with this common clinical problem and Allicin quickly

emerged as an agent worthy of testing in a standardized wound infection model. Using this

mouse model, we plan to follow wound progression in the presence of different levels of Allicin

applied at the wound site, and compare that to uninfected and untreated controls. We will follow

the progression of this infection in a number of ways: visually (by periodic photography of the

wound site), through the quantitative determination of inflammatory cytokine gene expression by

the mouse host, through the quantitative determination of virulence factor gene expression by the

pathogen, and histologic staining and microscopic analysis of wound tissue. These forms of

analysis will serve as the basis for determining whether Allicin is effective at controlling wound

infection and, if so, to perhaps yield important clues about the mechanism by which it operates.

These clues may be useful in designing more effective control on wound infection in the future.

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Abstract #12

Regulation of an Antibiotic-Induced Virulence Gene Cluster in Burkholderia pseudomallei Jennifer R. Klaus1, Patricia Silva1, and Josephine R. Chandler1

1Department of Molecular Biosciences, University of Kansas, Lawrence, KS, USA Abstract: The bacterium Burkholderia pseudomallei is a Category B select agent and opportunistic pathogen that causes the deep-lung disease melioidosis, an often-fatal and difficult-to-treat condition. The disease mechanisms of B. pseudomallei are poorly understood. Because B. pseudomallei is a BSL-3-restricted pathogen, studies on B. pseudomallei disease mechanisms are often carried out using a closely related non-pathogen, Burkholderia thailandensis. We are interested in the mal gene cluster, which has been shown to be important for virulence in several animal models. The mal genes are comprised of the polyketide biosynthesis genes malA-M, and malR. The DNA element containing the mal genes and malR is highly conserved in B. thailandensis and B. pseudomallei, and most of the Mal proteins have >80% sequence identity. MalR is a predicted member of the LuxR family of transcriptional regulators that are typically activated by acyl-homoserine lactone (AHL) signals. Although MalR has all of the conserved amino acids of AHL-binding LuxR-family proteins, we have shown that the MalR from B. thailandensis is an AHL-independent transcriptional activator of the mal genes. Expression of the mal genes is induced by certain antibiotics (e.g. trimethoprim) that activate MalR by driving malR expression. In standard laboratory conditions malR and the mal genes are not expressed. We have begun experiments to elucidate mal gene regulation in B. pseudomallei using an attenuated, BSL-2-approved strain. Our results show that in B. pseudomallei the mal genes and malR are activated by trimethoprim, which is clinically used to treat melioidosis infections. Our results suggest that the regulation of the mal cluster may be similar in B. thailandensis and B. pseudomallei. Our results also suggest the possibility that this virulence cluster may be activated by antibiotics during infections, which we aim to test in future experiments.

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Abstract #13 The Fatty Acid Kinase of Staphylococcus aureus Controls Virulence Christina N. Krute1 and Jeffrey L. Bose1

1Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS, USA

Abstract:

Virulence factor regulation in Staphylococcus aureus is under complex transcriptional

and post-transcriptional control. During a screen of the Nebraska Transposon Mutant

Library, we identified mutants in an operon encoding two hypothetical proteins, named

VfrA and VfrB, which had a dramatic decrease in α-hemolysin production. Further

analysis using non-polar mutations revealed that the second protein, VfrB, was the

primary contributor to conditionally-controlled α-hemolysin, protease production, and

expression of other secreted virulence factors. This regulation may be mediated by VfrB

activation of SaeRS, as α-hemolysin production in a vfrB mutant is restored in S. aureus

strain Newman, which encodes a constitutively active SaeRS system. Additionally,

expression of saeRS-controlled genes is down-regulated in vfrB mutants. In conjunction

with a role in virulence, VfrB was identified as a fatty acid kinase essential for uptake of

exogenous fatty acids. When proposed active site residues of VfrB are mutated, the

hemolytic activity of the enzyme is abolished, indicating a role for these residues in

kinase activity. Furthermore, it was discovered that VfrB works in cooperation with two

additional previously hypothetical proteins (termed FakB1 and FakB2). While VfrB

serves as a fatty acid kinase, FakB1 and FakB2 are fatty acid-binding proteins with

differing specificity. In vivo studies using a murine model of dermonecrosis revealed that

the vfrB mutant was hyper-virulent as observed by increased tissue damage, despite

similar numbers of bacteria present. Together, these studies identify a previously

undescribed fatty acid kinase complex found in gram-positive bacteria, and reveal the

connection between fatty acid metabolism and virulence factor regulation in S. aureus.

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Abstract #14

Mechanism of Vaccine-Induced Protection against Coxiella burnetii Lindsey E. Ledbetter1 and Guoquan Zhang1

1Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA Abstract: Coxiella burnetii is an obligate intracellular Gram-negative bacterium that causes the

zoonosis Q fever in humans. This disease manifests as an acute flu-like illness, although

it can escalate to a chronic and often fatal disease. It is also considered an occupational

hazard, therefore prophylactics should be studied to protect individuals who are at risk of

exposure to infected materials. There is currently a licensed whole-cell vaccine available,

however it is not approved for use in the United States due to adverse reactions in

previously sensitized individuals. Safe use of this vaccine requires multiple screening

procedures, which precludes a mass vaccination program. This whole-cell vaccine has

been shown to be protective against C. burnetii challenge, although the mechanism of

protection remains unknown. Previous work from our laboratory has demonstrated that

this protection is reliant on B cells. In addition, we have shown that these B cells produce

protective IgM independent of T cell help. Here we look further into the mechanism of

protection and show that it is specifically marginal zone B cells that proliferate upon

vaccination. This class of B cells is likely responsible for the majority of T cell-

independent IgM produced. Future studies will identify the innate cells responsible for

priming this adaptive immune response. By understanding the mechanism behind this

vaccine-induced protection, we will be able to develop safer, more effective vaccines.

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Abstract #15 Small-Angle X-ray Scattering Analysis of Bacteriophage Sf6 Structural Protein gp8: a Molecular Hub that Mediates the Assembly of the Tail Machine Lingfei Liang*, Tsutomu Matsui§, Thomas M. Weiss§, Haiyan Zhao* and Liang Tang* *Department of Molecular Biosciences, University of Kansas, Lawrence, KS, USA; §Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, USA Abstract:

Most double-stranded DNA (dsDNA) bacteriophages contain a highly specialized and

highly efficient tail structure responsible for attachment to host cells and injection of

phage DNA into host cytoplasm. The tail of dsDNA bacteriophage Sf6 is a multi-

protein, multi-functional molecular machine with a mass of 2.8 million Dalton and

consisting of 5 gene products present in as many as 51 subunits arranged with different

symmetries. Here we show that a tail component, gp8, of phage Sf6, exists in solution

in equilibrium of a monomer, a dimer and a hexamer. Small-angle X-ray scattering

(SAXS) analysis of the purified gp8 monomer shows a brick-shaped, globular protein

with a small protrusion. Fitting of the gp8 SAXS model into the electron cryo-

microscopy map of the entire tail machine defines molecular boundaries among gp8

monomers and with adjacent subunits of other tail components. Each gp8 monomer

interacts with two adaptor protein monomers, two head-binding domains and two

receptor-binding domains from two adjacent tailspike trimmers. The N-terminal

portions of the adaptor protein and the tailspike head-binding domain extend towards

gp8, which may generate additional intermolecular interactions. These interactions

unveil various symmetry mismatches and hold up the ultra-stable tail machine. These

results suggest that gp8 serve as a molecular hub that mediates the assembly of the tail

machine and integrates multiple functions.

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Abstract #16 Novel and Rapid Development of Stabilizers Against Infectious Disease Protein Toxins and Human Disease Alexandra Machen1, Pierce O’Neil1, and Mark T. Fisher1

1University of Kansas Medical Center / CHAPRx LLC, Kansas City, KS, USA Abstract: CHAPRx has developed a patent pending proprietary detection platform to rapidly and

rationally identify and validate protein stabilizers to prevent deleterious protein folding

reactions. We demonstrate that our accelerated platform can 1) validate small molecule

protein stabilizers to block protein toxin transitions during bacterial and viral infections,

2) reverse and prevent folding diseases, or 3) provide industry solutions to stabilize

protein therapeutics. The ability to directly and rapidly inhibit toxin transitions that

change folding shapes during infection has wide applications in human and animal

health. Our technology can rapidly identify and validate inhibitors of infection dependent

shape changes of numerous bacterial/viral protein toxins (e.g. Anthrax, Diphtheria,

Botulinum, Ricin, Tetanus, Shiga toxin, influenza, MRSA). Our technology platform can

rapidly develop stabilizers to prevent protein folding diseases. We have successfully

demonstrated stabilization against proteins of Parkinson’s disease, cystic fibrous, amyloid

diseases (transthyretin), breast cancer, bleeding disorders as well as in rare orphan

diseases such as Friedrich’s ataxia. The common thread between bacterial/viral protein

toxins and protein folding diseases is that they both undergo folding changes that then

result in disease.

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Abstract #17 NKG2D Genotype Alters Gut Microbiome Composition and Immunoglobulin A Production in The Large Intestine Kimberley N. Matocha1, Devin Koestler2 and Mary Markiewicz1

1The Department of Microbiology, Molecular Genetics & Immunology, University of Kansas Medical Center, Kansas City, KS; 2The Department of Biostatistics, University of Kansas Medical Center, Kansas City, KS Abstract: The NKG2D receptor and its ligands have been implicated in mediating the progression

of multiple autoimmune diseases including type I diabetes, colitis, rheumatoid arthritis,

chronic obstructive pulmonary disease, and a variety of cancers. Our lab uses the non-

obese diabetic (NOD) mouse model to ascertain the effects of NKG2D genotype on type

I diabetes and, more recently, its influence on the gut microbiome composition. In mice,

the NKG2D receptor is expressed on all natural killer cells, activated CD8+ T cells and T

cell subsets, CD4+ T cells, natural killer T cells, and gamma-delta T cells. Mouse-

expressed NKG2D ligands include retinoic acid inducible genes-1 (RAE1α-ε), minor

histocompatibility antigen (H60a-c), and mouse UL16-binding protein-like transcript

(MULT1). With the exception of intestinal epithelial cells, most healthy tissues do not

express NKG2D ligands. Given the unique property of NKG2D ligands being

constitutively expressed by intestinal epithelial cells, we investigated whether the

NKG2D genotype affects the gut microbiome composition. 16S Sequencing was

performed on a small portion of fecal pellet aseptically extracted from the colon.

Preliminary data suggests the gut microbiome composition is altered due to NKG2D

genotype. To further understand the mechanism with which NKG2D modifies the gut

microbiome composition, we assessed the role of mucosal-associated immunoglobulin A

(IgA). Via ELISA analysis, colonic fecal homogenate supernatants were analyzed for

IgA concentration and normalized to per gram feces. Our findings support NKG2D

genotype significantly alters IgA concentrations, specifically, in the large intestine thus

may prove to be quintessential for maintaining the gut microbiome composition.

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Abstract #18 The Induction of Small Colony Variant Staphylococcus aureus in Artificial Sputa Media Rachel McAllister1, Elizabeth Pascual1 and Erika Lutter1

1Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA Abstract: Cystic fibrosis (CF) is a genetic disease that results in increased mucus formation in lungs

and pancreas making individuals with CF prone to chronic infections by a number of

pathogens including Staphylococcus aureus. S. aureus is one of the first pathogens to

infect the respiratory tract of patients being the most prevalent pathogen in children and

early teens. Infection with S. aureus can persist into late adulthood. During infection

phenotypic variants, small-colony variants (SCVs), of S. aureus can emerge. SCVs are

believed to be caused by the selective pressure of antibiotics and are of great concern as

they have been increasingly isolated from CF patients. The aim of this study is to

determine if S. aureus isolated from CF patients grown in artificial sputum medium or

minimal media exhibit an altered SCV phenotype. It was found that growth of S. aureus

and prevalence of SCVs was enhanced when S. aureus was grown in artificial sputa

media compared to minimal media. This suggests conditions mimicking the environment

of the CF lung are more favorable for growth and SCV formation of S. aureus.

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Abstract #19 Probing the Kinetic Stability of Tetanus Neurotoxin Using Biolayer Interferometry along with a Novel Denaturant Pulse-Chaperonin Assay with Parallel Confirmation Via Transmission Electron Microscopy Pierce T. O’Neil1, Alexandra J. Machen1, Josh Burns2, Michael R. Baldwin2, and Mark T. Fisher1

1Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA; 2Department of Molecular Microbiology and Immunology, University of Missouri School of Medicine, Columbia, MO, USA Abstract: Tetanus neurotoxin (TeNT) is a virulence factor produced by Clostridium tetani. TeNT, synthesized as a single polypeptide, has three distinct domains: heavy chain C-terminus (HC), heavy chain N-terminus (HN) and the light chain (L). HC acts to target the toxin to neurons via acidic gangliosides and protein receptors. Correct receptor binding facilitates endocytosis, retrograde transport, and discharge into the spinal intersynaptic space. After HC-independent endocytosis into pre-synaptic inhibitory neuron endosomes, the pH decreases. The low pH triggers conformational change in the HN domain allowing it to insert into the endosomal membrane acting as a cation channel and translocon for the L chain. Once in the cytosol, the disulfide link between L and HN is reduced, and the L chain is now neurotoxic. To better understand the conformational change of HN associated with pH drop, we investigated the kinetic denaturation isotherm of the complete toxin. Using bio-layer interferometry (BLI), the toxin is attached onto a biosensor tip. This tip serves two purposes: 1) the toxin is oriented in the same manner to produce maximal signal, and 2) the toxin cannot interact with each other, thus preventing aggregation, a common hurdle when studying membrane toxins with exposed hydrophobic regions. Once the His-tagged TeNT is immobilized on the Ni-NTA biosensors, it is briefly exposed to urea. Following the urea pulse, the tip is submerged into a GroEL solution. GroEL, a bacterial chaperonin, ubiquitously binds to any exposed hydrophobic patches. Kinetic stability isotherms are generated by plotting urea concentration against GroEL binding signal. In parallel, we varied the pH (6.5-7.5) of the urea to see a shift in the isotherm curve. Using negative stain transmission electron microscopy, the TeNT-GroEL complexes were confirmed. This was achieved by releasing TeNT from the biosensor directly onto glow discharged grids into microliter volumes. Using the images captured, potential regions of instability were noted. With greater knowledge of conformational change, we hope to provide insight into directed design for inhibitors against TeNT. Any identified compounds can be tested using this methodology, where any leftward shift in the resulting isotherm and/or diminished GroEL binding indicates small molecule stabilizers of the toxin. Ultimately, we hope the inclusion of small molecules leads to novel anti-toxin drug therapies.

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Abstract #20

Bioinformatic Analysis of Antibiotic Resistance Mechanisms of Coxiella burnetii Nicholas P. Olivarez1, Guoquan Zhang1,2

1Department of Molecular Microbiology & Immunology, University of Missouri, Columbia, MO, USA; 2Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA Abstract: The obligate intracellular bacterial pathogen Coxiella burnetii is the causative agent of

the zoonotic disease, Q Fever. This pathogen is extremely resistant to environmental

damage and is easily transmitted by aerosolized particles from fluids of infected hosts.

Current therapeutic procedures for acute infections requires two weeks of treatment with

tetracycline class antibiotics; for chronic infections antibiotic use for a minimum of

eighteen months is required to resolve infections. Despite being the focus of continuous

research for over eighty years, little is known of the mechanisms behind this antibiotic

resistance, specifically, whether resistance is attributed to how the antibiotics interact

with the bacterial targets or if the acidic environment of the intracellular vacuole that the

bacteria replicate within plays a role in reducing the efficacy of antibiotics. To address

this, a thorough bioinformatic analysis of the putative antibiotic resistance proteins of C.

burnetii are compared to antibiotic resistance mechanisms of other well characterized

pathogens. This approach has established novel relationships at the protein level between

C. burnetii and well known pathogens. This work facilitates the modeling of predicted

protein structures of this important pathogen of humans and livestock to identify unique

domains on antibiotic resistance proteins that can better inform the development of

therapeutics by minimizing the duration of antibiotic treatment while maximizing

treatment efficacy.

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Abstract #21 Amino Acids T45 and R53 in Yersinia pestis YopK Putative Binding Region Suppress Host Inflammasome Response Rachel M Olson1,2, Kristen N Peters1,2, Deborah M Anderson1,2 Veterinary Pathobiology, University of Missouri, Columbia, MO, USA 2Molecular Microbiology and Immunology, University of Missouri, Columbia, MO, USA Abstract:

Yersinia pestis infection has caused an estimated 200 million deaths in multiple pandemics worldwide. Plague has been recognized as a re-emerging infectious disease with naturally occurring antibiotic resistance. Coupled with its potential for aerosol transmission and high mortality rate, this makes Y. pestis a dangerous potential bioterrorism agent. Yersinia species have a virulence plasmid encoding a type III secretion system (T3SS), which translocates effectors to subvert host defenses. The T3SS itself is sufficient to activate the inflammasome. YopJ, YopM and YopK are known to manipulate the inflammasome, but their relationship to each other is unclear. While deletion of yopJ or yopM has a very small effect on virulence, the deletion of yopK produces a severe virulence defect. YopK is a 21kDa protein with no known sequence homologs. We recently showed that YopK was involved in promoting YopJ-dependent macrophage apoptosis, suggesting YopK either has direct effector function or modulates the activity of YopJ and perhaps other Yop effectors. In this work, we studied this hypothesis by constructing a yopK-yopJmutant. We show that this strain causes a significant increase in IL-1β secretion by macrophages and dendritic cells compared to deletion of either yopJ or yopK alone, as well as WT and T3SS null bacteria. This result suggests that YopK prevents inflammasome activation caused by one or more elements of the T3SS. To further understand the potential role of YopK on inhibiting this host response, native or point mutant YopK-expressing plasmids were added back to the yopJ-yopK mutant. The suppression of IL-1β by YopK was dependent on amino acids T45 and R53 in the putative binding region of YopK. The critical amino acid for other known YopK functions is D46, suggesting multiple binding surfaces of YopK or a novel host target for the suppression of the inflammasome response. Together the results support a model whereby YopK is required to modulate host cell death responses during Y. pestis infection that is necessary to block protective inflammatory responses.

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Abstract #22 Thermostability of Alphaviruses Can Be Virus-Specific So Lee Park1,2, Yan-Jang S. Huang1,2, Susan M. Hettenbach2, Stephen Higgs1,2, and Dana L. Vanlandingham1,2 1Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS, USA; 2Biosecurity Research Institute, Kansas State University, Manhattan, KS, USA Abstract: Alphaviruses are important veterinary and human pathogens with significant epidemic

potential and disease burden. Whilst infections can cause incapacitating clinical diseases

and symptoms, detecting asymptomatic infections by laboratory diagnosis is critical for

disease surveillance prior to the epidemics. Serological assays are important methods for

laboratory diagnosis of alphavirus infections. Currently, plaque reduction neutralization

test (PRNT) is considered as the gold standard for serological assays because of the

demonstration of neutralizing capacity in serum samples. Heat inactivation of serum

samples at 56oC for 30 minutes is a required procedure before the neutralization tests to

eliminate complement activity and adventitious viruses. Although such method is

effective for the complete inactivation of flaviviruses, such parameters have recently been

demonstrated to be insufficient for Western equine encephalitis and chikungunya viruses.

Residual viruses can yield false negative PRNT results and represent a risk to laboratory

personnel. Despite its significance in diagnostic results and laboratory safety, it is not

known whether or not this high thermotolerance is a phenotype shared by all

alphaviruses. In this study, the thermostability profiles of Ross River, Barham Forest,

chikungunya, and o’nyong-nyong viruses were examined to determine the optimum

parameters for complete heat inactivation of these alphaviruses.

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Abstract #23 Activation of Dendritic Cells by the Subunit Vaccine for Shigella Melissa Pressnall1, Francisco Martinez-Becerra1, Olivia Arizmendi-Perez2, Wendy Picking1, William Picking1. 1Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA; 2Department of Molecular Biosciences, University of Kansas, Lawrence, KS, USA Abstract: Shigellosis is a disease with severe global impact yet it has no approved vaccine. This

gastrointestinal disease effects ~90 million people per year with 100,000 deaths primarily

in young children in developing countries. Shigella infects humans by utilizing a type

three secretion system (T3SS) which is highly conserved over the ~50 Shigella serotypes.

The T3SS works like a needle and syringe that injects bacterial protein effectors into the

host epithelial cells in the colon to promote bacterial entry. The internalized bacteria then

lyse the resulting phagosome which allows bacterial replication in the host cell

cytoplasm. Two proteins, IpaB and IpaD, control the activity of the T3SS from the tip of

the needle. Using a lethal pulmonary mouse challenge model, we have shown that

vaccination with IpaB + IpaD or a genetic fusion of these proteins (the DB Fusion) along

with the adjuvant dmLT is protective against a Shigella flexneri challenge as well as a

heterologous challenge by Shigella sonnei. In addition, we found that using both antigens

(either combined or in the DB Fusion) gives rise to higher cytokine activation and

protection profiles. We are working to determine the protective mechanism of the DB

Fusion vaccine and to identify the pathway for antigen presentation. The first step in this

effort is to analyze the effect of these proteins in antigen presenting cells such as

dendritic cells. In this study, we are stimulating dendritic cells with IpaB, IpaD,

IpaB+IpaD, or the DB Fusion, with or without dmLT to measure subsequent cytokine

release as well as the up-regulation of activation markers. Identification of the activation

markers and cytokines involved will help to describe a plausible mechanism for the

response of dendritic cells incubated with our candidate vaccine proteins.

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Abstract #24 Constitutive Expression of the NKG2D Ligand RAE1ε Within Pancreatic Islets Ameliorates Autoimmune Diabetes Development in Non-Obese Diabetic Mice Neekun Sharma1, Mary Markiewicz1 1Department of Microbiology, Molecular Genetics, & Immunology, University of Kansas Medical Center, KS, USA Abstract: Type 1, or autoimmune, diabetes is driven by T cell-mediated destruction of the insulin-

producing β cells in pancreatic islets. The natural killer cell activating receptor NKG2D

and its ligands are implicated in this process. However, the mechanism by which

NKG2D-NKG2D ligand interaction affects diabetes development is poorly understood.

Transgenic expression of the NKG2D ligand retinoic acid early transcript 1ε (RAE1ε) in

β-islet cells of the pancreas induces the recruitment of cytotoxic T lymphocytes (CTLs)

to the islets in mice. Surprisingly, however, we report here that the severity of

spontaneous autoimmune diabetes is decreased in non-obese diabetic (NOD) mice

expressing RAE1ε in islets (RIP-RAE1ε NOD). Despite enhanced infiltration of

lymphocytes to the pancreas, NKG2D expression is reduced on the surface of T cells

within the pancreas of RIP-RAE1ε NOD mice compared with control NOD mice. This

suggests that transgenic expression of RAE1ε in pancreatic islets leads to the down-

regulation of NKG2D surface expression on T cells, decreasing the ability of the cells to

respond to NKG2D engagement by ligands naturally expressed in the pancreas. We

further demonstrate that the NKG2D ligand H60a is expressed on T cells within the

pancreas of wild-type NOD mice. Together, these data suggest that interaction between

NKG2D and H60a expressed on T cells within the pancreas plays a critical role in the

development of NOD diabetes and that blocking this interaction may be therapeutically

useful in type 1 diabetes.

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Abstract #25 Protective Roles for NKG2D in NOD Diabetes Development Andrew P. Trembath1, Neekun Sharma1, Kimberly N. Matocha1, Mary A. Markiewicz1

1 Department of Microbiology, Molecular Genetics & Immunology, University of Kansas Medical Center, Kansas City, KS, USA

Abstract: Type I diabetes (TID) is a debilitating autoimmune disorder which requires ongoing insulin injections, and is associated with numerous long-term health consequences. This disease is mediated by autoreactive T cells that destroy the insulin-secreting β-islet cells within the pancreas. While it is clear that many genes contribute to the development of type 1 diabetes, its cause remains unknown. Both human and murine studies implicate the activating immune receptor natural killer group 2 member D (NKG2D) in enhancing TID development. However, the mechanism by which NKG2D signaling influences diabetes development has been unclear. Here we report that NKG2D signaling influences spontaneous autoimmune diabetes development in non-obese diabetic (NOD) mice via multiple, opposing mechanisms. First, we found that NKG2D influenced diabetes development via alteration in the microbiota composition. Second, when we eliminated this microbiota effect, we observed an unexpected protective role for NKG2D in NOD diabetes development. To determine how this effect was mediated, we examined NOD pancreatic islets for expression of NKG2D ligands and found that T cells within the pancreas of pre-diabetic NOD mice expressed the NKG2D ligand H60a. In vitro we observed both H60a and NKG2D on activated NOD CD8+ T cells. This suggested a possible role for NKG2D-ligand engagement during homotypic T cell-T cell interaction during both CD8+ T cell differentiation into cytotoxic T cells (CTLs) and during CTL effector function. We tested this hypothesis in vitro using CD8+ T cells from mice with various NKG2D expression levels and blocking antibodies against H60a. During differentiation, NKG2D-H60a interaction influenced the CTL effector cytokine response, but not cytotoxic granule release. In contrast, NKG2D-H60 interaction during the CTL effector phase did not affect cytokine production, but increased cytotoxic granule release. These results are the first to describe a role for NKG2D-ligand engagement via homotypic T cell-T cell interactions. Our findings suggest there are two roles for these interactions in NOD CTL responses. First, there is a dominant protective role for these interactions during NOD CD8+ T cell differentiation into CTL within pancreatic lymph nodes, second, there is a detrimental role for these interactions within the pancreatic islets where they increase β-cell killing.

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Abstract #26 Toward Understanding the Structural Basis of Inhibition of the Classical and Lectin Pathways of Complement by S. aureus Extracellular Adherence Protein Jordan L. Woehl1, Brandon L. Garcia1, Kasra X. Ramyar1, John K. Walker2, Michal Hammel3, Brian V. Geisbrecht1 1Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS, USA; 2St. Louis University School of Medicine, St. Louis, MO, USA; 3Lawrence Berkeley National Lab, SIBYLS High Throughput SAXS, Berkeley, CA, USA Abstract: The pathogenic bacterium Staphylococcus aureus actively evades many aspects of human innate immunity by expressing a series of secreted inhibitory proteins. A number of these proteins have been shown to inhibit the complement system. We recently reported that S. aureus Eap inhibits the classical and lectin pathways of the complement cascade by a previously undescribed mechanism (Woehl et al (2014) J. Immunol.). Specifically, Eap binds with nanomolar-affinity to complement C4b, and thereby blocks binding of the Classical and Lectin pathway pro-protease, C2. This effectively eliminates formation of the CP/LP C3 proconvertase, which is required for downstream complement activity. Although Eap’s mechanism of action has been demonstrated, little is known about how Eap interacts with complement C4b at the structural level. The full-length, mature Eap protein consists of four ~97 residue domains, which adopt a similar beta-grasp fold, and are connected through a short linker region. Through multiple assays, we have identified the 3rd and 4th domains as being critical for interacting with C4b and inhibiting the complement cascade. Co-crystallization efforts of Eap34 bound to a large fragment of C4b yielded ordered crystals, but they diffracted X-rays poorly; this has hampered our ability to obtain a satisfactory structural model. We therefore explored an alternative approach that takes advantage of the abundance of surface exposed lysines in the Eap protein. Through this lysine-acetylation footprinting technique, we have tentatively mapped and identified two key areas in Eap domains 3 and 4 that show protection upon being bound to C4b. Further experiments are underway to test this provisional identification of the C4b-binding site within Eap domains 3 and 4.

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14th Annual Great Plains Infectious Disease Meeting University of Kansas - Lawrence Kansas

November 6 - 7, 2015

MEETING REGISTRANTS

Last Name First Name Email Address Institution Affiliation Adam Phil Padam@kdheks.gov Kansas Dept. of Health Adamovicz Jeffrey adamoviczj@missouri.edu University of Missouri Adams Paige apadams@ksu.edu Kansas State University Agarwal Sanjeev s803a783@ku.edu KU Al-Murrani Sam murrani@babylonbioconsulting.com Prommune, Inc. Amachawadi Raghavendra agraghav@vet.ksu.edu Kansas State University Anderson Deborah andersondeb@missouri.edu University of Missouri Arizmendi Olivia olivia.arizmendi@ku.edu University of Kansas Ashitey Pearl p326a245@ku.edu University of Kansas Ault Kevin kault2@kumc.edu University of Kansas

Medical Center Barta Michael Barta.michael@ku.edu University of Kansas Bearson Brad brad.bearson@ars.usda.gov USDA; ARS; National

Laboratory for Agriculture and the Environment

Behar Amanda amanda.behar@okstate.edu Oklahoma State University

Berkland Cory berkland@ku.edu University of Kansas Bhattacharya Somanon sb2p3@mail.umkc.edu University of Missouri

Kansas City Bodugam Mahipal mahipalbiict@gmail.com University of Kansas Bose Jeffrey jbose@kumc.edu University of Kansas

Medical Center Bourne Christina cbourne@ou.edu University of Oklahoma Brunelle Brian brian.brunelle@ars.usda.gov National Animal Disease

Center Carter Wayne wcarter@kclifesciences.org Kansas City Area Life

Sciences Institute Carvalho Claudia cmdasilvacarvalho@fhsu.edu Fort Hays State

University Chandler Josephine jchandler@ku.edu University of Kansas Chen Kangming kangming@ksu.edu Kansas State University Cherla Rama cherlar@missouri.edu College of Veterinary

Medicine; University of Missouri-Columbia;

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Choi Seong-O sochoi@ksu.edu Kansas State University Coate Eric eackr3@mail.missouri.edu University of Missouri-

Columbia Coyle Kathryn kathryn.coyle@uky.edu University of Kentucky Davis Heather hfrisch@k-state.edu Kansas State University Dean Bartholomew bdean@ku.edu university of Kansas DeDonder Keith keith@mvsinc.net Veterinary and

Biomedical Research Center, Inc.

DeDonder Sarah sarahdedonder@gmail.com Kansas Department of Agriculture

DeKosky Brandon brandon.dekosky@nih.gov NIAID Dhariwala Miqdad modcp6@mail.missouri.edu University Of Missouri-

Columbia Dowdell Alexander adowdell@kumc.edu University of Kansas

Medical Center Durand-Heredia

Jorge jmdt4b@mail.missouri.edu University of Missouri at Columbia

Eleshy Rawan rawan.eleshy@okstate.edu Oklahoma State University

Eshelman Kate kleshelman@ku.edu University of Kansas Esquivel Brooke bdp3k9@mail.umkc.edu University of Missouri-

Kansas City Fairlamb Max m999f013@kumc.edu University of Kansas

Medical Center Fisher Mark mfisher1@kumc.edu University of Kansas

Medical Center Ganguly Arghya arghya@ku.edu University of Kansas Gao Philip gao@kanpro-research.com KanPro Research Inc. Garcia Brandon garciab@ksu.edu Kansas State University Geanes Eric esggg7@mail.umkc.edu University of Missouri-

Kansas City Gelhaus Carl cgelhaus@mriglobal.org MRIGlobal Hawley Blake blake@iahglobal.com Integrated Animal Health Hefty Scott pshefty@ku.edu University of Kansas Hermanas Timothy tmh3m8@mail.missouri.edu University of Missouri at

Columbia Hettenbach Susan smhett@bri.ksu.edu Kansas State University Hilliard Kinsey kahnr9@mail.missouri.edu University of Missouri Horvat Rebecca rhorvat@kumc.edu University of Kansas

Med Center Hsieh Hsinyeh hsiehh@missouri.edu University of Missouri Hulangamuwa Wasundara cwhulang@ksu.edu Kansas State University Hustead David david.hustead@boehringer-

ingelheim.com Boehringer Ingelheim Vetmedica

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Jafarain Sohaila sohailaj@vet.ksu.edu Kansas State University Javed Salim javeedsalim@ku.edu University of Kansas Jha Jay jayjha@ku.edu University of Kansas Jimenez Wanda wpjimenez@mail.fhsu.edu Fort Hays State

University Johnson Lauren lxjohnson1@wichita.edu Wichita State University Jordan Lorne lornejordan@ku.edu University of Kansas Kamath Divya dvkf57@mail.umkc.edu University of Missouri-

Kansas City Keleher Lauren keleherl@missouri.edu University of Missouri Klaus Jennifer jennklaus@ku.edu University of Kansas Krute Christina ckrute@kumc.edu University of Kansas

Medical Center Kumar Prashant prashant.kumar@ku.edu University of Kansas Lane Abigail Abigail.Lane07@gmail.com Kansas State University Ledbetter Lindsey lindseyeledbetter@mail.missouri.edu University of Missouri-

Columbia Liang Lingfei l152l176@ku.edu University of Kansas Lin Chungho linchu@missouri.edu University of Missouri Lutter Erika erika.lutter@okstate.edu Oklahoma State

University Ma Wenjun wjma@ksu.edu Kansas State University Machen Alexandra amachen@kumc.edu KUMC Manley Michael mmanley@kumc.edu KU Medical Center Markiewicz Mary mmarkiewicz@kumc.edu University of Kansas

Medical Center Marks Kayla kdmfhc@mail.missouri.edu University of Missouri-

Columbia Martinez Becerra

Francisco francisco.martinez@ku.edu University of Kansas

Massa Nicole nmassa2015@ku.edu The University of Kansas Matocha Nicole kmatocha@kumc.edu The University of Kansas

Medical Center McAllister Rachel rachel.mcallister@okstate.edu Oklahoma State

University McDonald Dave david.mcdonald@wichita.edu Wichita State University McGill Jodi jlmcgill@vet.k-state.edu Kansas State University McKinney Megan mem2015@ku.edu University of Kansas Miriyala Nagaraju nagarajumiriyala@ku.edu University of Kansas Monteiro-Riviere

Nancy nmonteiro@ksu.edu Kansas State University

Moore Kaitlin kkmoore2@mail.fhsu.edu Fort Hays State University

Moral Mario mmoral@ku.edu University of Kansas

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Nicholson Eric eric.nicholson@ars.usda.gov National Animal Disease Center; USDA

Nicholson Tracy tracy.nicholson@gmail.com USDA Nowak Martha msnowak@ksu.edu K-State Olathe Olivarez Nicholas nicholas.olivarez@mail.missouri.edu University of Missouri Olson Rachel RMO34B@MAIL.MISSOURI.EDU University of Missouri-

Columbia ONeil Pierce poneil@kumc.edu University of Kansas

Medical Center Park So Lee parksolee@vet.k-state.edu Kansas State University Parsel Suzanne tbogina@ksu.edu Kansas State University

Olathe Patel Ami patelam@missouri.edu Laboratory for Infectious

Disease Research Paudyal Anuja paudyalanuja1@gmail.com Fort Hays State

University Picking Wendy wendy.picking@ku.edu University of Kansas Picking William picking@ku.edu University of Kansas Pratt Miranda mjpratt@ku.edu The University of Kansas Pressnall Melissa pressnallm@ku.edu University of Kansas Qiu Iris qiux@ku.edu The University of Kansas Rezac DJ dj@mvsinc.net Veterinary and

Biomedical Research Center, Inc.

Rivera Mario mrivera@ku.edu University of Kansas Riviere Jim jriviere@ksu.edu Kansas State University Rosa-Molinar Eduardo erm@ku.edu The University of Kansas Sease Rebecca davis.a.rebecca@gmail.com Kansas State University Sharma Neekun nsharma4@kumc.edu University of Kansas

Medical Center Stein Sydney steinsr@health.missouri.edu University of Missouri Stewart George stewartgc@missouri.edu University of Missouri Taylor Anna Marie Amhump@k-state.edu Kansas State University Trembath Andrew atrembath@kumc.edu University of Kansas

Medical Center Vanlandingham Dana dlvanlan@vet.ksu.edu Kansas State University Vishwakarma Vikalp vikalp@ku.edu University of Kansas Wakefield Susan swakefield@ku.edu University of Kansas Ward Claire ceward@ksu.edu Kansas State University Whitaker Neal nealwhitaker@ku.edu University of Kansas Woehl Jordan jlwoehl@ksu.edu Kansas State University Yao Huili hly@ku.edu University of Kansas Zhang Xuehan liuxuehan1996@hotmail.com Kansas State University Zhang Guoquan zhangguo@missouri.edu University of Missouri

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