The PSWRCE Protein and PCR MS Core will collaborate on the following projects:
Specific Aim i - Proteomics Analyses for Burkholderia, Coccidioides, and Reservoir Vaccines Projects ? Identify protein biomarkers for use as diagnostic or vaccine candidates (from serum, urine, and lung homogenates, with depletion of abundant mouse/human proteins as necessary) ? Perform comparative proteomics on pre-fractionated samples (e.g., extracellular proteins or cell wall proteins of two different strains of an organism, or wild type vs. mutant) ? Build """"""""blood meal"""""""" protein library to use to identify host vertebrate proteins in tick nymphs Specific Aim 2 - Development of Diagnostic Assays ? Use biomarker IDs of Sp.
Aim i to develop bioaffinity MS for pathogen identification ? Use MS to guide development of targeted ELISA or other fluorescence assays (characterize expressed protein antigens, antibodies, and labeled antibodies by MS throughout development of assay) ? Compare antibodies vs. small ligands for capture of antigens in bioaffinity MS Specific Aim 3 - Ibis TSOOO PCR-MS Biosensor Applied to Reservoir Vaccines, Burkholderia &Coccidioides ? PCR MS (on mitochondrial DNA) for ID of vertebrate host on which vector has fed (e.g., tick nymph fed on mule deer) and ID of tick-borne pathogenic organism present ? PCR MS for rapid ID and strain typing of pathogens, as well as confirmation of the presence of toxin genes, antibiotic resistance genes, selected virulence factor genes Specific Aim 4 - Characterization of Protein Complexes (uncompensated synergistic activity) ? Structurally characterize large protein-protein complexes (e.g., pili involved in virulence, assemblies of bacteriophage tail fiber proteins useful for biosensors) using a one-of-a-kind modified QTOF MS The antigen discovery coupled to support of diagnostic assay development, PCR-MS, bioaffinity MS, and structural characterization of large protein-protein complexes are unique capabilities of the core.
Collaborations of the Core with RCE projects will allow: identification of proteins that will be used in diagnostic tests or as vaccine candidates;determination of protein expression differences between virulent and non-virulent strains of organisms;identification of vertebrate hosts of tick vectors, allowing disease transmission to be better tracked;pathogens to be rapidly characterized by PCR MS;and characterization of large protein complexes important in virulence or diagnostic design.
|Hertz, Tomer; Beatty, P Robert; MacMillen, Zachary et al. (2017) Antibody Epitopes Identified in Critical Regions of Dengue Virus Nonstructural 1 Protein in Mouse Vaccination and Natural Human Infections. J Immunol 198:4025-4035|
|Marques, Adriana R; Yang, Xiuli; Smith, Alexis A et al. (2017) Citrate Anticoagulant Improves the Sensitivity of Borreliella (Borrelia) burgdorferi Plasma Culture. J Clin Microbiol 55:3297-3299|
|Parameswaran, Poornima; Wang, Chunling; Trivedi, Surbhi Bharat et al. (2017) Intrahost Selection Pressures Drive Rapid Dengue Virus Microevolution in Acute Human Infections. Cell Host Microbe 22:400-410.e5|
|Bortell, Nikki; Flynn, Claudia; Conti, Bruno et al. (2017) Osteopontin Impacts West Nile virus Pathogenesis and Resistance by Regulating Inflammasome Components and Cell Death in the Central Nervous System at Early Time Points. Mediators Inflamm 2017:7582437|
|Barbour, Alan G (2017) Infection resistance and tolerance in Peromyscus spp., natural reservoirs of microbes that are virulent for humans. Semin Cell Dev Biol 61:115-122|
|Nualnoi, Teerapat; Norris, Michael H; Tuanyok, Apichai et al. (2017) Development of Immunoassays for Burkholderia pseudomallei Typical and Atypical Lipopolysaccharide Strain Typing. Am J Trop Med Hyg 96:358-367|
|Nakajima, Rie; Escudero, Raquel; Molina, Douglas M et al. (2016) Towards Development of Improved Serodiagnostics for Tularemia by Use of Francisella tularensis Proteome Microarrays. J Clin Microbiol 54:1755-65|
|Zeltina, Antra; Bowden, Thomas A; Lee, Benhur (2016) Emerging Paramyxoviruses: Receptor Tropism and Zoonotic Potential. PLoS Pathog 12:e1005390|
|Ziegler, Christopher M; Eisenhauer, Philip; Bruce, Emily A et al. (2016) A novel phosphoserine motif in the LCMV matrix protein Z regulates the release of infectious virus and defective interfering particles. J Gen Virol 97:2084-9|
|Park, Arnold; Yun, Tatyana; Hill, Terence E et al. (2016) Optimized P2A for reporter gene insertion into Nipah virus results in efficient ribosomal skipping and wild-type lethality. J Gen Virol 97:839-43|
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