The
purpose of this project is to develop new antimycobacterial agents for control
and/or eradication of mycobacterial diseases. This project involves
collaborations with a Crystallography Group and an Organic Chemistry group.
This is a drug-design program based upon recombinant DNA technology, X-ray
crystallography, organic synthesis and site-directed mutagenesis.
Sponsor: NIAID/NIH
PIs: William W. Barrow, Rebecca J. Morton
Oklahoma Medical Research Foundation: Allen B. Edmundson, Philip
Bourne
SRI, Birmingham, Alabama: William J. Suling, Robert Reynolds
This
is a collaborative project with Southern Research Institute in Birmingham,
Alabama. It is designed to
investigate the antimycobacterial activity of purine analogs against Mycobacterium
tuberculosis.
It involves organic synthesis, purine metabolism and in vitro assays of purine
derivatives against M. tuberculosis.
Sponsor:
NIAID/NIH
PIs:
William B.
Parker
Department
of Veterinary Pathobiology: William W. Barrow, Joseph A. Maddry
Shipping
Fever: New Approaches to Understanding Prevention and Management
The objective is to determine infectious agents and host responses that cause respiratory disease and/or defend cattle from disease. Naturally occurring cattle diseases at Noble Foundation, privately owned, USDA, and OSU herds are being investigated. Emphasis is primarily on bovine viral diarrhea virus infection and pasteurellosis. Vaccination programs are being evaluated and new recommendations given.
Sponsor: Noble Foundation
PIs: Anthony W. Confer, S. Mady
Dabo, Robert W. Fulton, Jerry W. Ritchey
Department
of Veterinary Clinical Sciences: John G. Kirkpatrick, Robert A. Smith
This
project determines changing patterns, geographical differences, risk factors,
and management practices related to bovine respiratory disease. The influence
of various bacteria and viruses is studied. In addition, the pharmacokinetics
and efficacy of newer therapies and new-generation vaccines are evaluated. The
host-pathogen relationship is characterized at the molecular level.
Sponsor: Oklahoma Agricultural Experiment Station
PIs: A. W. Confer, R. W. Fulton, R. J. Panciera, K. D. Clinkenbeard, R.
J. Morton
Mannheimia haemolytica Outer Membrane Protein PlpE:
Characterization of Epitopes Stimulating Homologous and Heterologous Serotype
Protection
This project is a molecular and immunologic approach to
studying an immunologically important outer membrane protein of M.
haemolytica. It compares
the PlpE protein from serotypes 1, 2, and 6 with respect to important epitopes
for immunity.
Sponsors: USDA
CSREES, National Research Initiative Competitive Grant
PIs: A.
W. Confer, Sahlu Ayalew
Mannheimia
haemolytica Bacterin-Toxoid Efficacy Studies
This
project studies the efficacy of commercial M. haemolytica vaccines in an experimental
challenge model.
Sponsor:
Pfizer
Animal Health
PIs:
A. W.
Confer, R. J. Panciera
Pasteurella
(Mannheimia) haemolytica Lipopolysaccharide-Leukotoxin Complex
A
protein exotoxin termed leukotoxin (LKT) and an endotoxin termed
lipopolysaccharide (LPS) produced by Pasteurella haemolytica are the most important
virulence factors in the causation of shipping fever in cattle. LKT and LPS are
chemically distinct toxins.
Likewise, the biological activities of these toxins can also be
distinguished. Although LKT and LPS have separate toxic activities, evidence of
cooperative toxic effects has been recognized. We recently determined that LPS
is complexed with LKT. The hypothesis for this proposal is that: LKT catalyzes
the transfer of aggregated LPS to specific receptors on target cells, which
mediates the LPS intoxication. The specific objectives are: 1) To compare
target cell activation elicited by LPS·LKT complex with that for
LPS-free LKT, isolated LPS, and non-complexed LKT + LPS to determine whether
LKT facilitates LPS target cell activation, and 2) To identify receptors
mediating LKT facilitated LPS target cell activation. For bovine neutrophils, naturally complexed LPS·LKT induced higher expression
of TNF-a
than either purified LKT or LPS individually. Anti-CD18 Mab blocked LKT induced
TNF-a
expression, but not that of purified LPS or LPS·LKT complexed, suggesting
that a receptor other than CD18 is involved in LPS or LPS·LKT complexed signaling in
bovine neutrophils.
Sponsor: United States Department of
Agriculture, National Research Initiatives Competitive Grants Program
PI: Ken Clinkenbeard
Pasteurella haemolytica produces an exotoxin termed
leukotoxin (LKT) that has been implicated as an important virulence factor in
shipping fever pneumonia in cattle. LKT specifically intoxicates ruminant
leukocytes and platelets, but LKT has limited or no effect on leukocytes from
other species. Bovine CD18 acts as the species-specific and leukocyte-specific
receptor for P. haemolytica LKT. Prior to exposure to LKT, CD18 was found to be evenly
distributed around the periphery of the bovine leukocyte plasma membranes.
Following one minute of exposure, LKT and CD 18 were found associated with
patches of degenerative plasma membranes. No CD18 or LKT was observed in
regions of intact plasma membranes. This supports LKT-induced capping of CD18.
The rapid degeneration of the associated plasma membrane in these patches
suggests that CD18 localization to these regions may play a role in loss of
plasma membrane integrity.
Sponsor: Oklahoma Agricultural
Experiment Station
PI: Ken Clinkenbeard
The
objective of this program is to expand the suite of sensors for the
Environmental Systems Management, Analysis, and Reporting neTwork (E-SMARTâ) to include devices for the
detection of chemical and biological warfare agents. This program involves assessing the state-of-the-art
detection technology for these materials, with an emphasis on Former Soviet
Union (FSU) technology. The scope
of the program involves assessment of pertinent detection technology,
organization and direction of research, the development, testing, and
evaluation of emerging detection technology, and the validation of operation
through the integration and testing of the devices with the E-SMART
demonstration system currently in place at Tinker AFB in Oklahoma City,
Oklahoma. Device validation will
include testing with stimulants and live agents at certified facilities. System validation will be conducted at
Tinker AFB using approved materials, remote signal transmissions, and/or simulated
signal inputs to demonstrate detection and response actions. Since the actual technologies to
be integrated are not known at this time, a technical discussion and cost
estimate (in the cost proposal) for technology anticipated as likely candidates
for E-SMART integration is provided.
Sponsor: Air Force Research
Laboratory and General Atomics
PIs: Ken
Clinkenbeard, Rebecca
Morton
Department
of Physiological Sciences: Carey Pope
The
objective of this program is to expand the suite of sensors for the
Environmental Systems Management, Analysis, and Reporting neTwork (E-SMARTâ) to include devices for the
detection of biological warfare agents. Specifically, polymerase chain reaction
(PCR) devices will be assessed as well as detection technology developed by
program support to Former Soviet Union (FSU) under the Enhancement of E-SMART
for Chemical and Biological Warfare Agent Detection project.
Sponsors: Air Force Research
Laboratory, General Atomics
PIs: Ken
Clinkenbeard, Rebecca
Morton
Networked
Terrorism Detection System
The
overall goal of this proposal is to develop highly specific approaches for
detecting and identifying explosives, nerve gases and BW agents using a unique
amplifying fluorescent polymer (AFP) which will greatly increase the speed and
sensitivity of detection of explosives and CBW agents and to apply this
development to produce a continuous real-time microarray networkable detection
system for use against terrorist threats. Central to the overall goal is the
underlying hypothesis that AFP can be used as a sensor platform and
functionalized with specific probes to detect in continuous real-time
explosives and specific CBW agents at minimal effective levels. This hypothesis
will be tested for nitroaromatic and other explosives, a nerve gas agent, and
three BW agents. Once developed, the functionalized AFPs will be incorporated
into a microarray sensor. Individual microarray sensors can be placed in a
networked system such as General Atomics E-SmartÒ such that high value
buildings or complexes can be continuously monitored for these and other CBW
agents. Therefore, the research conducted for this project will: 1) develop
probes specific and selective for explosives and prototype CBW agents, 2)
functionalize AFPs for detection of prototype agents, 3) produce and test a
prototype AFP microarray sensor for explosives and CBW detection, and 4) conduct
refinement, application, and commercialization of our MIPT AFP microarray sensor.
Sponsors: Oklahoma City Memorial for
the Prevention of Terrorism, National Institute of Justice
PIs: Ken
Clinkenbeard, Rebecca
Morton, John Wyckoff
Department
of Physiological Sciences: Cyril Clarke, Carey Pope, Jerry Malayer
Biological
Warfare Agent Water Monitor
The
threat from biological warfare (BW) agents in combat and terrorist scenarios
makes the ability for rapid detection and identification of BW agents of great
importance. Oklahoma State University, along with their commercial partner
Nomadics, Inc., proposes to adapt amplifying fluorescing polymer (AFP)
technologies currently being developed for BW agent detection in aerosols for
monitoring of water supplies for BW agents. Such a monitor is of great interest
to the Joint Services Agent Water Monitor (JSAWM) project of the Army Soldier
Biological and Chemical Command (SBCCOM). Hurdles for development of
near-real-time continuous monitoring of water supplies for BW agents are
requirements for high sensitivity and specificity for up to 20 agents monitored
simultaneously with minimal use of consumable reagents. The approach proposed
herein is novel using new technologies that can interrogate water supplies with
a highly sensitive regenerable reagent adaptable for numerous BW agents. The
project plan is to demonstrate the utility of this approach by developing a
prototype portable BW agent water monitor for two BW agents, Francisella
tularensis,
the causative agent of tularemia, and the water-borne pathogen
enterohemorrhagic Escherichia coli O157:H7 (EHEC). Central to the overall goal is the underlying hypothesis that AFP can be used as a
sensor platform for BW agents in water supplies and functionalized with
specific probes to detect in near-real-time BW agents at minimal infectious
levels. This hypothesis will be tested for 2 BW agents. Once developed, the
functionalized AFPs will be incorporated into a recoverable microbead (MB)
prototype monitor. Our objectives are to: 1) Functionalize AFP for oligonucleotide-based detection
of BW agent Francisella tularensis and Escherichia coli O157:H7, 2) Adapt available technologies for disruption of BW
agent in water supplies for presentation of genomic and ribosomal
oligonucleotides for sensing, 3) Develop functionalized AFPs into
specific-sized microbeads, and 4) Develop a prototype BW agent water monitor
for F. tularensis and E. coli O157:H7.
Sponsors: Oklahoma Center for
Advancement of Science and Technology, Oklahoma Applied Research Support
PIs: Ken
Clinkenbeard, Rebecca
Morton
Department
of Physiological Sciences: Jerry Malayer
Amplifying
Fluorescent Polymer Diagnostics for Respiratory Pathogens
The
Phase I solicitation topic is aimed at development of rapid diagnostic tests
for early detection of emerging respiratory pathogens. The United States Navy
has interest in this topic, which largely arose out of concern about the
increasing incidence of acute respiratory disease amongst military trainees
subjected to overcrowding, stress, and frequent mobilization to disease-endemic
areas (Naval Health Research Center, San Diego). Changes in pathogen susceptibility, lack of adult vaccines,
reemergence of problem pathogens, and the inadequacy of current diagnostic
methods, especially in the military field environment, emphasized the need for
sensitive, specific, rapid, and robust detection methods. Nomadics’ Phase I proposal
addresses development of a signal amplification technology (amplifying
fluorescent polymer, or AFP) that will enable early detection of a bacterial
target via binding to a specific aptamer probe. The target chosen for initial development and testing of the
technology is Bordetella pertussis lipopolysaccharide (LPS).
Phase I technical objectives include: 1) Modification of amplifying
fluorescent polymer (AFP) for attachment of a quencher-labeled oligonucleotide
recognition (aptamer) sequence, 2) Testing and optimization of transduction
between the aptamer-functionalized AFP film and a target molecule via a binding
event. Initially transduction will
be established and optimized using a recently published aptamer beacon
(Hamaguchi et al., 2001) selected specifically for thrombin identification, and 3)
Selection of highly specific and sensitive aptamers targeted to LPS of B.
pertussis.
Sponsors: Office of Naval Research,
Small Business Innovative Research
PIs: Ken
Clinkenbeard
Nomadics:
Jean Clarke
Development
of Aptamer Beacons to Lipopolysaccharide for the Real-Time Sensing of BW Agents
Force
protection is of utmost importance, but a lack of real-time sensing
technologies for biological warfare (BW) agents leaves US forces vulnerable to
conventional as well as terrorist’s BW attacks. The major barriers to
real-time sensing of BW agents are 1) a lack of robust probes for detection of
BW agents, and 2) inadequate sensitivity of sensor platforms through which
target probes transduce their detection signals. We propose to conduct a “proof-of-concept”
project to marry a new type of probe technology with exceptional robustness
termed aptamers with a newly conceived sensing platform with exceptional
sensitivity termed AFP beacons to overcome the current barriers for developing
real-time BW sensing. Aptamers are highly stable and specific oligonucleotides
that work like monoclonal antibodies (Mab) to bind directly to BW target
agents. However, unlike Mab, aptamers have exceptional stability even under
field conditions. Aptamer technology is only beginning to be applied to
detection of infectious agents. We propose to develop aptamers to the highly
agent-specific O-antigen of lipopolysaccharides of a prototype BW agent,
enterohemorrhagic E. coli, to demonstrate that aptamers can be used to specifically
detect BW agents. In addition, we will engineer our anti-LPS aptamers to act as
beacons. That is, when the aptamer binds to its target it will switch from an
“off” to an “on” signal by turning on specific
fluorescence directly. No consumable reagents are required. Once DEPSCoR
proof-of-concept project is accomplished we will transition to prototype sensor
development using this technology.
Sponsor: Army Research Laboratory
PI: Ken
Clinkenbeard
Identification
of P. multocida Surface Protein(s) with Specific Binding to Chicken Intestinal
Secretory Immunoglobulin A
Fowl
cholera involves infection via the respiratory route during outbreaks. However, the digestive tract is also
severely affected during outbreaks. This project will identify vaccine
candidate proteins in Pasteurella multocida that bind to chicken
secretory IgA (sIgA) involved in immunity to the bacteria.
Sponsor:
OSBA
PIs:
S. Mady
Dabo, Anthony W. Confer, John H. Wyckoff III
Baboon Research Resource Program
This project will study ways
of enhancing the well-being of baboons in both breeding and research settings,
develop a number of reagents for quantitation of various components of the
immune system of baboons, and assess viral pathogens indigenous in baboons and
the biology of these infectious agents.
Sponsors: National Center for Research
Resources, National Institutes of Health
PIs: Richard Eberle
Oklahoma University Health
Science Center: G. White, J. Wallis
This project provides support
for the derivation of a Specific Pathogen Free breeding colony of baboons that
are free of six herpes viruses and four retroviruses. As part of this project
new, more sensitive immunoassays and PCR tests for detection of these baboon
viruses will be developed.
Sponsors: National Center for Research
Resources, National Institutes of Health
PIs: Richard Eberle
Molecular Diagnostic Tests for Simian Herpesviruses
This project provides support
for the molecular characterization of monkey B virus. The data derived from
these molecular studies are then used to develop new assays for the detection
of B virus infections in humans and macaque monkeys, and to differentiate
infections in humans caused by B virus versus the human herpes simplex viruses.
Sponsors: National Center for Research
Resources, National Institutes of Health
PI: Richard Eberle
Bovine
Viral Diarrhea Disease Virus (BVDV) Vaccines: Antibody Response to Heterologous BVDV Strains
The study will determine the range of
heterologous immunity in calves receiving modified live virus (MLV) or killed
BVDV vaccines. Currently there are two recognized antigenic types of BVDV, Type
1 and 2. The study will determine if these vaccines induce antibodies to
various Type 1 and 2 viruses.
Sponsors: Grand
Laboratories, Inc., Pfizer Animal Health
PIs: Robert W. Fulton,
Anthony W. Confer
Genetic
and Antigenic Variability of BVDV in Cattle Infections
Bovine viral diarrhea viruses (BVDV)
isolates from the Oklahoma Animal Disease Diagnostic laboratory (OADDL) will be
obtained from clinical/necropsy cases.
The viruses will be typed as BVDV 1a, 1b, or 2. Potentially there will be additional
typing and/or groups. Field
isolates from naturally occurring disease including persistently infected (PI)
cattle will be compared with vaccinal stains and standard reference
strains. A phylogenetic survey of
the BVDV subtypes from the field isolates, vaccinal strains and reference strains
will be performed to detect relationships between the viruses and their genetic
stability. Neutralization tests
will be performed to compare the subtypes to the vaccinal strains. Potentially new subtypes may warrant
additional subtypes in the vaccines.
Sponsor: Oklahoma Agricultural
Experiment Station
PIs: Robert W. Fulton, A. W.
Confer
Oklahoma Animal Disease Diagnostic
Laboratory: J.T. Saliki
Antibiotic Administration and Vaccination
with Live Bacterial Vaccine in Calves
This study will determine if an antibiotic
given to calves that have been administered avirulent Pasteurella
haemolytica and Pasteurella multocida
vaccine will decrease the immune responses to the immunogens. Calves will
receive MicotilÒ
antibiotic and Once PMHÔ Pasteurella
haemolytica and P. multocida vaccine. The calves’
sera will be tested for P. haemolytica and P.multocida
antibodies.
Sponsor: ELANCO
Animal Health
PIs: Robert W. Fulton, Anthony
W. Confer
Bovine viral diarrhea viruses (BVDV) occur
as biotypes, cytopathic (CP) and noncytopathic (NCP), and as genotypes, 1 and
2. Certain BVDV disease forms occur with different biotypes/genotypes. The
molecular differences among biotypes/genotypes will be investigated by PCR and
nucleic acid sequencing. Virulence markers of BVDV will be investigated.
Sponsor: Oklahoma
Agricultural Experiment Station
PIs: Robert W. Fulton, Jean M.
d’Offay, Anthony W. Confer, Jerry W. Ritchey
Oklahoma Animal Disease Diagnostic Laboratory:
Jeremiah T. Saliki
Evaluation
of Viral Vaccine Containing Infectious Bovine Rhinotracheitis Virus (IBRV),
Bovine Viral Diarrhea Virus 1 and 2 (BVDV), Parainfluenza-3V (PI-3V), and
Bovine Respiratory Syncytial Virus (BRSV) in Preventing Infection and
Respiratory Disease in Cattle
The
purpose of the study will be to determine if pre-weaning vaccination of ranch
calves with viral vaccine: 1) reduces respiratory disease, and 2) reduces
transmission of viruses in calves moved from auction markets and commingled
with the fresh calves under feedlot conditions.
Sponsor:
Fort Dodge
Animal Health
PIs: Robert Fulton, A.W. Confer
Oklahoma
Animal Disease Diagnostic Laboratory: J.T. Saliki
Department
of Veterinary Clinical Sciences: D.L. Step
This
is a cooperative study on the role of Amblyomma maculatum in the spread of Hepatozoon
americanum
to dogs from other as-yet-unknown vertebrate hosts.
Sponsor:
Krull
Endowed Professorship in Veterinary Parasitology
PIs:
S. A. Ewing,
Roger J. Panciera
This
is a study of the development of veterinary parasitology from the classical
discipline of parasitology in North America, especially the role of Wendell H.
Krull, founding head of the veterinary parasitology discipline at Oklahoma
State University.
Sponsor:
Krull
Endowed Professorship in Veterinary Parasitology
PI:
S. A. Ewing
Canine
hepatozoonosis, a serious, often fatal disease of dogs caused by the protozoan
parasite, Hepatozoon americanum, was recently recognized to occur with considerable frequency in
Oklahoma. Our studies have identified the vector tick, many aspects of the
reproductive cycle of the parasite in the domestic dog and in the vector tick,
and established phylogenetic relationships of H. americanum to certain other protozoan.
Ongoing research addresses studies of the pathogenesis of skeletal lesions of
the disease, and development of a serologic test to be used for diagnosis and
for epidemiological investigations designed to identify potential reservoirs
available to infect the vector tick.
Efforts are being made to validate a serologic test developed at OSU by
acquiring serum samples from veterinarians in a wide geographic area.
Sponsors:
College of
Veterinary Medicine, Krull Endowed Professorship in Veterinary Parasitology
PIs:
Roger J.
Panciera, S.A. Ewing, A. Alan Kocan, Terry W. Lehenbauer
Oklahoma
Animal Disease Diagnostic Laboratory: Jeremiah T. Saliki
Reduced
Disease Transmission by Tick Control on Rodents
This
study investigates the feasibility of reducing tick-transmitted diseases by
developing a safe but effective medicated food/bait for wild mammals that will
inhibit immature tick development, thus reducing the potential number of tick
disease vectors.
Sponsor: Oklahoma Center for the
Advancement of Science and Technology
PIs:
A. Alan
Kocan
Department
of Entomology: Robert Barker
Determination
of the Susceptibility of Coyotes to Infection with Babesia gibsoni
Experimental
infections in captive coyotes will be compared to those initiated with the same
isolate in domestic dogs to determine the potential role of coyotes as a
natural reservoir for this parasite.
Sponsor:
College of
Veterinary Medicine
PIs:
A. Alan
Kocan, Jim Meinkoth
Characterization
of Cats Surviving Cytauxzoon felis Infections
This
study investigates the clinical and biological aspects of cats that have
survived naturally occurring infections with Cytauxzoon felis. Studies will focus on
genetic characterization of the non-fatal isolate and attempted
tick-transmission trials.
Sponsors:
College of
Veterinary Medicine, Morris Animal Foundation
PIs:
A. Alan
Kocan, Jim Meinkoth
A
Cell Culture-Derived Vaccine for Anaplasmosis
Anaplasma
marginale
harvested from tick cell culture will be tested as an antigen for a new and
improved vaccine for bovine anaplasmosis. This antigen should result in the
development of a vaccine that is safe, easily standardized, and free of
contaminating bovine cells and pathogens. A vaccine dose will be formulated and
tested in cattle for vaccine efficacy.
Sponsor: Novartis Animal Vaccines,
Inc.
PIs: Katherine M. Kocan, Edmour F.
Blouin, Jose de la Fuente
Novartis
Animal Health: Thomas Halbur, Virginia C. Onet
A Cell Culture-Derived
Vaccine for Anaplasmosis
Anaplasma
marginale
harvested from tick cell culture will be tested as an antigen for a new and
improved vaccine for bovine anaplasmosis. This antigen should result in the
development of a vaccine that is safe, easily standardized, and free of
contaminating bovine cells and pathogens. A vaccine dose will be formulated and
tested in cattle for vaccine efficacy.
Sponsor: Oklahoma Center for the
Advancement of Science and Technology
PIs: Katherine M. Kocan, Edmour F.
Blouin, Jose de la Fuente
Novartis Animal Health:
Thomas Halbur, Virginia C. Onet
A Cell Culture-Derived
Vaccine for Bovine Anaplasmosis
Anaplasma
centrale will
be propagated in tick cell culture.
Organisms harvested from cell culture will be tested as a live vaccine
for bovine anaplasmosis in South Africa and Israel. The cell culture derived A.
centrale
should result in the development of a vaccine that is safe, easily
standardized, and free of contaminating bovine cells and pathogens. A vaccine
dose will be formulated and tested in cattle for vaccine efficacy.
Sponsor:
U.S.- Israel
Cooperative Development Research (CDR) Program
PIs: Katherine M. Kocan, Edmour
F. Blouin, José de la Fuente
Varda Shkap (Israel), Eric
Zweygarth (South Africa)
Prevalence
of Escherichia coli O157:H7 in Stocker Cattle
This project is being conducted at the
Willard Sparks Beef Research Center and will provide an estimate of prevalence
for Escherichia coli O157:H7 infection in stocker cattle and the influence of
receiving programs on its persistence.
This information will be useful for developing other projects aimed at
preventing or reducing this infection, which is a significant food borne
pathogen in humans.
Sponsor:
College of
Veterinary Medicine
PIs: Terry W. Lehenbauer
Department
of Animal Science: Stan Gilliland
Department
of Veterinary Clinical Sciences: Robert A. Smith
Genetic Construction of
Bovine CD18 that Resists P. Haemolytica Leukotoxin
Pasteurella (Mannheimia)
haemolytica is
the cause of an economically important respiratory disease of cattle known as
“shipping fever.” The purpose of this project is to investigate the
relationship between a toxin (leukotoxin, LKT) secreted by the Pasteurella bacterium and bovine immune
cells. The main objective of this
project is to definitively identify the bovine cell receptor that LKT uses to
bind and mediate toxicity.
Following identification of the receptor, the specific binding domain
will be characterized. The
information generated from this work will lead to improved treatment regimens,
vaccine strategies, and possibly the development of cattle genetically
resistant to “shipping fever.”
Sponsor: U. S. Department of
Agriculture
PI: Jerry W. Ritchey
Murine
Model for Monkey B Virus Infection
Non-human primates are an
integral part of biomedical research programs, particularly as animal models of
human disease. As long as monkeys
are used for research, zoonotic infection by simian viruses will continue to be
an occupational hazard. Human
infections by monkey B virus, a herpes virus of macaques involve the central
nervous system and left untreated are usually fatal. The goal of this project is to establish and characterize,
both pathologically and immunologically, a murine model for B virus infections. Establishment of a well characterized
murine model system for B virus infection will serve as an important resource
in which hypothesis-driven studies can be performed such as evaluation of
anti-viral drugs, challenge assessment of potential vaccines, and investigation
of the role of various viral genes/proteins in determining the pathogenic
properties of these viruses.
Sponsor:
National
Institutes of Health
PI:
Jerry W.
Ritchey
Functional Genomic/Proteomic Analysis of Campylobacter spp.
Campylobacter infection, generally
food-borne, is a significant global public health problem, representing the
leading cause of enteritis in humans. As with most food-borne diseases, the
molecular basis for the mechanisms of Campylobacter infection is poorly
understood. As the total genomic sequence of C. jejuni subsp. jejuni has recently become
available, the next major step will be to investigate the function of the two
genes/proteins revealed by the genome project and their expression under in
vivo
conditions. A key environmental signal for controlling bacterial pathogenesis
is the availability of iron in the host. This project proposes to investigate
the iron regulon of C. jejuni, using DNA microarrays and proteome analysis. The data from this
study will be used to construct knockout mutations in selected genes that
appear to be environmentally regulated; the knockout strains will then be
tested in a stringent in vivo model of Campylobacter infection, to assess whether these genes are
essential for Campylobacter pathogenesis.
Cytokine
Expression in Response to Brucella Vaccines
Cytokine responses induced by
Brucella polysaccharides
in blood cells from vaccinated- and unvaccinated-cattle and -mice,
unvaccinated-pigs and -humans were evaluated in vitro. Reverse transcriptase-polymerase chain reaction was used to
detect cytokine mRNA. Development
of a cytokine diagnostic profile indicative of induction of immunity for
application to human vaccinates was attempted.
Sponsor: Canada Department of National
Defense
PI: John H. Wyckoff III
Synthetic peptides with
sequences homologous to Mycobacterium bovis proteins will be used to
develop better test reagents for field and laboratory diagnosis of bovine
tuberculosis. Cattle immunized with M. bovis will be compared for
responsiveness to these antigens and those of M. avium to determine both specific
and cross-reactive responses.
Sponsor: Oklahoma Agricultural
Experiment Station
PIs: John H. Wyckoff III
Department of Biochemistry
and Molecular Biology: Richard Essenberg
Heat shock protein-specific T
lymphocytes derived from Brucella abortus vaccinated cattle will be characterized by flow
cytometry for surface marker expression and cytotoxic function against infected
monocyte-derived macrophages. Cytokine production by the T lymphocytes will be
analyzed through RT-PCR. These studies will define a host defense effector
mechanism against brucellosis.
Sponsor: U. S. Department of
Agriculture
PIs:
John H.
Wyckoff III, Anthony W. Confer