Research

The Great Lakes Regional Center of Excellence supports different types of laboratory research, reflecting a variety of approaches to fighting disease. Some projects are ambitious, multi-year studies, pooling the best resources in the region through collaboration across institutions and disciplines. Other projects employ an intense focus on a smaller area of research for a shorter period of time, with the hope that the results could quickly impact public health. Other projects aim to enlist the best young scientists into biodefense and emerging infectious disease research.

Research Projects

Research Projects in the Great Lakes RCE are multi-year, multi-disciplinary studies aimed at the development of new vaccines or therapies against diseases caused by agents that can be used as biological weapons. Research Projects provide the backbone of scientific study in the consortium. The following diseases are studied in GLRCE Research Projects:

Research Project 1: Anthrax

B. anthracis vegetative cells (DNA stained red) infect a human fibroblast monolayer (fibroblast DNA stained green). This image was collected on a Nikon TE-2000E inverted microscope coupled to a spinning-disk confocal unit.

Bacillus anthracis, the bacteria that causes anthrax, is a feared biological weapon due to its ability to be distributed through the air and to cause a high percentage of fatal disease, as demonstrated in the letter attacks on the United States in the fall of 2001. Though the infection can be treated with antibiotics, strains that have been engineered to be resistant to current antibiotic therapy are known to exist. Thus the discovery of new drugs to treat anthrax is a high biodefense priority.

Research Project 1 focuses on the development of new antibiotic therapy against anthrax. Drugs discovered in this project may also be effective against other infectious bacteria as many pathogens share common disease pathways. The scientists conducting this research are an interdisciplinary team of microbiologists, computational biologists, structural biologists and biochemists, and come from several institutions, including the University of Chicago, University of Michigan and Argonne National Laboratory.

Research Project 2: Botulism

Botulinum toxin is the most poisonous substance known to man. It is so potent that only one gram of toxin, evenly dispersed and inhaled, has the potential to kill more than one million people. Both easily obtained and grown, Botulinum toxin is believed to have been developed as a biological weapon by several countries, including the former Soviet Union and Iraq.

The Botulinum toxin affects nerve cells, and, if untreated, can cause paralysis and death. Treatment options consist primarily of antibodies to the toxin, which are only effective if administered early; antibodies can prevent additional toxin action but cannot reverse existing paralysis. There is currently no licensed vaccine that protects against botulism intoxication, nor any drug therapy to inhibit the action of the toxin. Research Project 2 employs a multi-disciplinary approach to develop a botulism vaccine and new anti-toxin drugs for therapy. Investigators with expertise in microbiology, biochemistry, physiology, and medicine from the University of Wisconsin at Madison, the Medical College of Wisconsin and the University of Illinois at Urbana-Champaign have teamed to achieve these goals.

Research Project 4: Plague

Immunofluorescent detection of the F1 capsule of Yersinia pestis. F1 capsule was detected using anti-F1 rabbit polyclonal serum as primary antibody. Cells were then labeled with Alexa Fluor 647 anti-goat anti-rabbit IgG.

Yersinia pestis is the bacterium that causes the plague or "black death", a disease that killed 250 million people in the middle ages and is responsible for more human mortality than any other infectious agent. Plague is a feared weapon of mass destruction because the bacteria can be disseminated in the air and transmitted from person to person, allowing for exponential spread of the highly fatal form of the disease, pneumonic plague.

There is currently no licensed vaccine in the United States, and though antibiotics are effective against this disease, they must be administered within 24 hours of the onset of symptoms to be effective (a timeframe that often precedes diagnosis of the infection). Microbiologists, immunologists and computational biologists from the University of Chicago, University of Illinois at Urbana-Champaign, Michigan State University and Argonne National Laboratory collaborate on Research Project 4 to develop safe and effective plague vaccines and post-exposure therapies.

Research Project 5: Ebola

Ebola virus-like particles. These filamentous-shaped particles - the hallmark of filoviruses - are produced in the lab. They resemble ebola virus, and allow the scientists to study the molecular mechanisms of the virus without the need for biosafety level 4 containment. (Photo credit: Y. Kawaoka)

Among the biological threats considered most serious are the viral hemorrhagic fevers. Ebola virus causes severe hemorrhagic fever in humans and non-human primates that can kill more than 80% of its victims. This threat is particularly formidable due to our limited knowledge of its pathogenesis and natural reservoir. Thus we currently have inadequate preventative and therapeutic measures making research aimed at protection against Ebola virus infections a high priority for the GLRCE.

Investigators at the University of Wisconsin at Madison are focused on the development of new vaccines and anti-viral drugs that will combat Ebola virus infections. Vaccinologists and virologists at the Canadian Science Center for Human and Animal Health and at the U.S. Army Medical Research Institute of Infectious Diseases in Maryland collaborate in this effort by providing the specialized facilities and reagents needed to study this important pathogen.

Research Project 6: Viral Hemorrhagic Fevers

The structure of West Nile Virus as determined by electron microscopy. (Photo credit: Science magazine)

Though less deadly than Ebola, Dengue hemorrhagic fever (DHF) virus accounts for over 99% of the cases of viral hemorrhagic fever worldwide. It is currently estimated that over 3 billion people live in areas at risk for Dengue hemorrhagic fever and thousands of people die each year from this disease. During each decade the number of DHF cases and their geographic distribution has increased steadily. This is due in part to the natural reservoir of this virus, the mosquito, which allows for high mobility and transmissibility of the virus.

Dengue virus is part of a large class of hemorrhagic fever viruses called flavivirus that also includes the West Nile and Yellow fever viruses. All are carried by insect vectors, such as the mosquito, thereby giving them the ability to spread rapidly and cause epidemics. Limited quantities of investigational vaccines and anti-viral drug therapies are available for some of the hemorrhagic fever viruses, but these the efficacy of these therapies has not been rigorously demonstrated. Therefore, research aimed at the development of new therapies and preventative measures is a high priority for the Great Lakes RCE. Investigators at Purdue University have designed a comprehensive, interdisciplinary approach to studying and developing new drug therapy against hemorrhagic fever viruses. Experts in microbiology, structural biology and drug design chemistry participate in this research.

Research Project 8: Designed Antibiotic Peptides and Mimetics

Scientists working on Research Project 8 at the University of Minnesota -- Twin Cities are studying bacterial membrane-disintegrating compounds as a possible means of controlling infectious disease. These novel compounds, which act by disrupting the integrity of the entire bacterial membrane, have considerable potential as therapeutics for biodefense. Research Project 8 is focused on developing new compounds and improving promising ones already discovered in their lab. These antibiotic agents are especially promising because their low molecular weights will ease the production, storage and use of the antibiotic as a dry powder.

Preliminary tests of the new antibiotic compounds are promising. While the compounds have not yet been shown to be effective against the NIAID Category A, B & C Priority Pathogens, they have been shown to be effective against closely related pathogens. The compounds have been shown to be protective against Bacillus anthracis (Sterne strain) and several clinical isolates of Staphylococcus aureus and E. coli.

The scientists of Research Project 8 are currently designing new compounds, optimizing the antibacterial activity of the compounds already developed, and assessing the biological efficacy of these compounds.

Research Project 9: Lung Innate Immune Responses to Franciselle Tularensis: A Central Role for the Macrophage

Five of the six Category A microbes, including F. tularensis (Ft), cause their most severe disease when inhaled as aerosols. Due to the daily bombardment of inhaled particulates, lung innate immune responses are unique, primarily driven by an immunoregulatory program that minimizes inflammatory damage in the alveoli. A critical mediator of this response is the alveolar macrophage (AM). For infectious agents that normally infect via the aerosol route, the lung appears to be relatively incompetent in controlling early microbial growth, particularly in the case of intracellular pathogens of macrophages. Ft is one such intracellular pathogen and is particularly threatening as a bioweapon for several reasons. First, the organism causes life-threatening infections after exposure to very small inocula making dissemination of Ft into the environment a serious concern. Second, it can cause disease by a number of infection routes, making prevention of infection in the event of an attack significantly more difficult. Third, the pneumonic form is associated with the greatest morbidity and mortality and aerosols can rapidly infect large populations. Finally, clinical conditions would most likely manifest as respiratory infections indistinguishable from influenza or other viruses.

To better understand virulence factors important during infection and the details of the host response which causes the disease, several research projects are currently in progress including:

1. Molecular determinants of phagocytosis of Ft and their regulation by lung collectins and other macrophage C-type lectins;


2. Characterization of the interactions of Ft with a new class of intracellular pathogen recognition molecules termed CATERPILLER proteins that are linked to the macrophage inflammasome;


3. Characterization of key macrophage biochemical pathways activated by Ft that regulate inflammatory responses; and


4. Macrophage responses and pathogenesis related to novel Ft acid phosphatases.

A greater understanding of Ft pathogenesis, with an emphasis on the unique immune response in the lung, will lead to new diagnostic, therapeutic and vaccine strategies.

Research Project 10: Bruccella Vaccine Development

Brucella is a human pathogen and bioterror agent for which no safe, effective human vaccine exists. Our ultimate goal is to develop immunization strategies against virulent Brucella. To achieve this goal, we are exploring two approaches:

1. Recombinant invasive E. coli and
2. Irradiated Brucella vaccine development.

First we are identifying immunogenic Brucella proteins for inclusion in the invasive E. coli vector.
Second, we have immunized mice with invasive E. coli vaccine vectors expressing the Brucella specific antigens and metabolically active/inactive irradiated Brucella.
Third, we are assessing efficacy of the Brucella antigen-expressing invasive E. coli and irradiated Brucella vaccines through clearance of Brucella challenged mice by in vivo imaging. Further, cytokine responses to the challenged mice are being assayed.
In conclusion, we believe our goal of developing and testing these novel vaccine strategies will contribute to achieving a safe, effective human vaccine against Brucella.