Bruce Steven Klein
University Of Wisconsin Madison
Project start date: 1995-01-01
Project end date: 2013-12-31
Sponsored Links Excellgen http://Excellgen.com
MOLECULAR PATHOGENESIS OF BLASTOMYCOSIS
Bruce Steven Klein, Professor
University Of Wisconsin Madison, 21 N. Park Street, Suite 6401, Madison, Wi 53715-1218
Grant 5R37AI035681-16 from National Institute Of Allergy And Infectious Diseases
Keywords: Active Sites; Adhesins, Bacterial; Amino Acid Substitution; Assay; Bacterial Adhesins; Binding; Binding (Molecular Function); Bioassay; Biologic Assays; Biological Assay; Biological Function; Biological Process; Blastomyces; Blastomyces dermatitidis; Blastomycosis; C-terminal; Cations, Divalent; Cell Surface Antigens; Controlled Study; Data; Divalent Cations; EF Hand Motifs; EF Hands; EGF; EGF gene; EGF-Like Domain; Engineering; Engineerings; Event; Fungi, Filamentous; Gene Transcription; Genes; Genes, LacZ; Genes, Regulator; Genetic Alteration; Genetic Change; Genetic Transcription; Genetic defect; Goals; Histoplasma capsulatum; Host Factor; Host Factor Protein; Immune; Immunity; Infection; Integration Host Factors; LacZ; LacZ Genes; Lung; Mediating; Molds; Molecular; Molecular Interaction; Morphogenesis; Mutate; Mutation; N-terminal; NH2-terminal; Pathogenesis; Pathogenicity; Pathogenicity Factors; Peptide Domain; Phase; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Protein Conformation; Protein Domains; Proteins; Purified Protein Derivative; Purified Protein Derivative of Tuberculin; RNA Expression; Regulator Genes; Reporter; Respiratory System, Lung; Role; Shapes; Sites, Active; Structure; Surface; Surface Antigens; Surface Markers, Immunologic; Surface Markers, Immunological; Tandem Repeat Sequences; Tandem Repeats; Tertiary Protein Structure; Transcription; Transcription, Genetic; Transcriptional Regulatory Elements; Tubersol; URG; Virulence; Virulence Factors; Work; Yeasts; adhesin; base; fungus; gene product; genome mutation; interest; mutant; pathogen; pulmonary; regulatory gene; response; social role; synthetic peptide; tool; trans acting element; tuberculin purified protein derivative
Project start date: 1995-01-01
Project end date: 2013-12-31
Budget start date: 1-JAN-2010
Budget end date: 31-DEC-2010
5R37AI035681-16 (2010): $377153
Grants awarded to Bruce Steven Klein
MECHANISMS OF ANTI-FUNGAL VACCINE IMMUNITY
Bruce Steven Klein, Professor
University Of Wisconsin Madison, 21 N. Park Street, Suite 6401, Madison, Wi 53715-1218
Grant 5R01AI040996-13 from National Institute Of Allergy And Infectious Diseases
Abstract: Vaccines are one of the most cost effective public health tools in human history. Several experimental vaccines to fungi are being explored, but have reached clinical trials and or are commercially available despite the increasing rates of fungal infections in healthy and immune deficient hosts. Vaccines to fungi demand a clear understanding of the mechanisms of adaptive immunity. This proposal addresses basic mechanisms of anti-fungal immunity by continuing to investigate a novel vaccine that we created against Blastomyces dermatitidis. In the last grant cycle, we began work on a genetically-engineered live attenuated vaccine. Vaccinated mice are uniformly protected against lethal pneumonia and acquire sterilizing immunity that is mediated chiefly by CD4 T-cells. Here, we propose to investigate the cellular and molecular mechanisms behind priming and long-term maintenance of vaccine induced CD4 T-cells. We have engineered the vaccine strain with model T-cell epitopes and bred the corresponding TCR transgenic mice so that we can carefully monitor and quantify pivotal stages during the induction of vaccine immunity and memory vaccine uptake by APC, antigen delivery to the draining lymph nodes, display of peptideMHC complexes, and priming and durability of memory CD4 T-cells at the clonal level. We hypothesize that vaccine yeast injected into the skin are captured by select dendritic cell (DC) subsets that migrate over several days to draining lymph nodes, where they move deep into the paracortex to prime naive T-cells. CD4 T-cells exert primacy in vaccine immunity in part because they out-compete CD8 T-cells for peptide MHC complexes on the DC surface and/or modulate the detour pathway of cross-presentation. CD4 T-cells establish a dynamic pool of memory subsets - TEM and TCM - in which TCM cells ultimately confer durable memory. Our specific aims are to (1) Elucidate the APC that engulfs vaccine yeast and presents it to naive CD4 T-cells in the draining lymph nodes. (2) Investigate the location and kinetics of antigen presentation within the draining lymph nodes, and the ensuing priming of CD4 T-cells and establishment of durable CD4 T-cell memory. (3) Define the mechanisms behind primacy of CD4 T- cells over CD8 T-cells during the induction of vaccine immunity. Findings from our studies will delineate the fundamental mechanisms of vaccine immunity and memory to fungi. These insights will foster the rational design of vaccines against fungal pathogens and other microbial agents
Keywords: APC; ATGN; Address; Adoptive Transfer; Animals; Antifungal Agents; Antifungal Drug; Antigen Presentation; Antigen-Presenting Cells; Antigenic Determinants; Antigens; Apoptosis; Apoptosis Pathway; Attenuated Live Virus Vaccine; Attenuated Vaccines; B Cell-Associated Molecule CD40; Binding Determinants; Blastomyces dermatitidis; Blastomycosis; Body Tissues; Breeding; CD4 Positive T Lymphocytes; CD4 T cells; CD4 lymphocyte; CD4+ T cell; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; CD40 Antigens; CD40 Type II Isoform; CD40 molecule; CD40L Receptor; CD8; CD8B; CD8B1; CD8B1 gene; CDw40 Antigen; Cell Death, Programmed; Cell Locomotion; Cell Migration; Cell Movement; Cell surface; Cells; Cells, CD4; Cellular Migration; Clinical Trials; Clinical Trials, Unspecified; Communicable Diseases; Complex; Confocal Microscopy; Cross Presentation; Data; Dendritic Cells; Dermal; Edodekin Alfa; Engineering; Engineerings; Epitopes; Epitopes, T-Lymphocyte; Event; FP593; Fostering; Funding; Fungal Vaccines; Fungicides, Therapeutic; Fungus Diseases; Grant; History; Human; Human, General; IL-12; IL12; Immune; Immunity; Immunologic Accessory Cells; In Situ; In Vitro; Infection; Infectious Disease Pathway; Infectious Diseases; Infectious Diseases and Manifestations; Infectious Disorder; Interleukin-12; Investigation; Kinetic; Kinetics; LYT3; Life; Live-attenuated Vaccine; Location; Lung diseases; Lymph node proper; Maintenance; Maintenances; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Memory; Mice; Mice, Transgenic; Microscopy, Confocal; Modeling; Molecular; Monitor; Monocytes / Macrophages / APC; Motility; Motility, Cellular; Murine; Mus; Mycoses; NKSF; Natural Killer Cell Stimulatory Factor; Nerve Growth Factor Receptor-Related B-Lymphocyte Activation Molecule; Pathway interactions; Patients; Peptide-MHC; Peptide-Major Histocompatibility Protein Complex; Peptide/MHC Complex; Peptides; Phase; Pneumonia; Pneumonitis; Property; Property, LOINC Axis 2; Proteins; Public Health; Pulmonary Diseases; Pulmonary Disorder; Pulmonary Inflammation; Receptor Signaling; Recording of previous events; Relative; Relative (related person); Respiratory Disease; Respiratory Disorder; Respiratory System Disease; Respiratory System Disorder; Reticuloendothelial System, Lymph Node; Role; Skin; Staging; Staining method; Stainings; Stains; Surface; T-Cell Epitopes; T-Cells; T-Lymphocyte; T-Lymphocyte Epitopes; T4 Cells; T4 Lymphocytes; TNFRSF5 Receptor; Tea; Testing; Thymus-Dependent Lymphocytes; Tissues; Transgenic Mice; Transgenic Organisms; Tumor Necrosis Factor Receptor Superfamily, Member 5; Vaccinated; Vaccination; Vaccine Design; Vaccines; Vaccines, Attenuated; Veiled Cells; Work; Yeasts; accessory cell; anti-fungal; antifungals; base; cell motility; chemokine receptor; clinical investigation; cost effective; cytokine; design; designing; drFP583; ds red protein; dsFP593; fungal infection; fungus; fungus infection; gene product; helper T cell; immunogen; in vivo; insight; live vaccine; lung disorder; lymph gland; lymph nodes; memory CD4 T cell; memory CD4 T lymphocyte; microbial; new vaccines; next generation vaccines; novel vaccines; pMHC; particle; pathogen; pathway; public health medicine (field); red fluorescent protein; social role; thymus derived lymphocyte; tool; transgenic; uptake
Project start date: 1997-05-01
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5R01AI040996-13 (2010): $610399
5R01AI040996-12 (2009): $603224
TARGETING HYBRID HISTIDINE KINASE FOR BROAD SPECTRUM ANTIFUNGAL THERAPY
Bruce Steven Klein
University Of Wisconsin Madison, 21 N. Park Street, Suite 6401, Madison, Wi 53715-1218
Grant 5RC1AI086025-02 from Office Of The Director, National Institutes Of Health
Abstract: This challenge grant addresses the broad challenge area (03) "Biomarker Discovery and Validation". The challenge topic is 03-AI-101, entitled "Identification, characterization and evaluation of novel pathogen or host targets that may lead to the development of antimicrobials with broad-spectrum activity." The title of the proposal is Targeting hybrid histidine kinase for broad-spectrum antifungal therapy. Despite the growing prevalence of severe fungal infections, we have a paucity of anti-fungal drugs, most of them are fraught with toxicity due to cross-reactivity with shared mammalian host targets, and only one new class of anti-fungal has been developed in the last decade. This proposal is designed to capitalize on a recent path-breaking discovery that a hybrid histidine kinase regulates global control of dimorphism and virulence in fungi. We hypothesize that hybrid histidine kinase represents an ideal target for the development of anti- fungal drugs to treat patients. This new target offers many features that the field has been seeking for development of broad-spectrum anti-fungal drugs, since it is highly conserved throughout the fungal kingdom and is not present in the human genome. We provide strong preliminary data supporting the concept and feasibility of this novel drug target. We have in place a strong drug discovery infrastructure for developing anti-fungal drugs, with campus leaders who are co-investigators on this grant, offering expertise in key areas and facility support, ranging from fungal and chemical biology, medicinal pharmacy, and drug lead optimization involving in vitro and in vivo animal studies. Our approach offers a powerful complimentary strategy that provides a cell based reporter system for identifying compounds that require the signaling pathway for their mode of action (aim 1), together with a pharmacophore 3-D model to find compounds that directly act on these targets in pathogenic fungi of divergent genera throughout the kingdom (aims 2 and 3). We expect that this work will ultimately lead to anti-fungal drugs that target multiple steps in this pathway, and are effective at treating a broad range of serious fungal infections in humans and perhaps plants. The number of fungal infections has risen dramatically in the United States over the last 10 years, and has reached the top 10 in causes of annual mortality in part because of the lack of efficacious anti-fungal drugs. This proposal addresses that public health need by seeking to identify, characterize and capitalize on a novel drug target in pathogenic fungi for the development of effective new anti-fungal drugs
Keywords: 3-D; 3-D structure; 3-Dimensional; 3-dimensional structure; 3D structure; Address; Agriculture; Americas; Animals; Antibiotic Resistance; Antifungal Agents; Antifungal Drug; Antifungal Therapy; Area; Aspergillus; Aspergillus fumigatus; Assay; Azole resistance; Azole resistant; Bacteria resistance; Bacteria resistant; Bacterial resistant; Binding; Binding (Molecular Function); Bioassay; Biologic Assays; Biological Assay; Biology; C. albicans; C.albicans; Cancers; Candida; Candida albicans; Caspofungin; Cell Communication and Signaling; Cell Signaling; Cells; Cessation of life; Chemicals; Chemistry; Communicable Diseases; Congresses; Data; Data Banks; Data Bases; Databank, Electronic; Databanks; Database, Electronic; Databases; Death; Development; Development and Research; Disease; Disorder; Docking; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drug Therapy; Drugs; EC 2.7; Evaluation; Food; Fungi, Filamentous; Fungicides, Therapeutic; Fungus Diseases; Galactosidase; Generalized Growth; Goals; Grant; Growth; Hand; High Throughput Assay; Human; Human Genome; Human, General; Hybrids; In Vitro; Individual; Industrial fungicide; Infection; Infectious Disease Pathway; Infectious Diseases; Infectious Diseases and Manifestations; Infectious Disorder; Infrastructure; Institution; Intracellular Communication and Signaling; Investigators; Killings; Kinases; Lead; Libraries; Malignant Neoplasms; Malignant Tumor; Mammalian Cell; Mammals, Mice; Man (Taxonomy); Man, Modern; Medication; Mice; Microbe; Modeling; Molds; Molecular; Molecular Interaction; Monilia; Mortality; Mortality Vital Statistics; Murine; Mus; Mycoses; Pathway interactions; Patients; Pb element; Pharmaceutic Preparations; Pharmaceutical Agent; Pharmaceutical Preparations; Pharmaceuticals; Pharmacies; Pharmacologic Substance; Pharmacological Substance; Pharmacotherapy; Pharmacy facility; Phosphotransferases; Plants; Plants, General; Pneumonia; Pneumonitis; Prevalence; Public Health; Pulmonary Inflammation; R & D; R&D; Reaction; Reporter; Research Infrastructure; Research Personnel; Researchers; Resistance; Resistance to antibiotics; Resistance, Antibiotic; Resistant to antibiotics; Role; Running; S cerevisiae; Saccharomyces cerevisiae; Science of Chemistry; Sepsis; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Societies; Stress; Structure; System; System, LOINC Axis 4; Temperature; Testing; Tissue Growth; Toxic effect; Toxicities; Toxicity Testing; Toxicity Tests; Transphosphorylases; United States; Validation; Virulence; Work; Yeast, Baker`s; Yeast, Brewer`s; Yeasts; agricultural; anti-fungal; anti-microbial; antibiotic resistant; antifungals; antimicrobial; bacterial resistance; base; biological signal transduction; biomarker; bloodstream infection; chemical genetics; clinical data repository; clinical data warehouse; cross reactivity; cytotoxic; data repository; design; designing; dimorphism; disease/disorder; drug discovery; drug/agent; fludioxonil; fungal infection; fungicidal; fungicide; fungus; fungus infection; glucan synthase; heavy metal Pb; heavy metal lead; high throughput screening; histidine kinase; in vitro Assay; in vitro activity; in vivo; inhibitor; inhibitor/antagonist; insight; malignancy; mammalian genome; neoplasm/cancer; novel; ontogeny; overgrowth bacterial; pathogen; pathogenic bacteria; pathway; pharmacophore; protein-histidine kinase; public health medicine (field); public health relevance; relational database; research and development; resistance to Bacteria; resistance to Bacterial; resistance to azole; resistant; resistant to Bacteria; resistant to Bacterial; resistant to azole; response; sensor; small molecule; small molecule libraries; social role; three dimensional structure; treatment of fungal infectious disease; virtual
Relevance: The number of fungal infections has risen dramatically in the United States over the last 10 years, and has reached the top 10 in causes of annual mortality in part because of the lack of efficacious anti-fungal drugs. This proposal addresses that public health need by seeking to identify, characterize and capitalize on a novel drug target in pathogenic fungi for the development of effective new anti-fungal drugs
Project start date: 2009-09-26
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1AI086025-02 (2010): $495552
1RC1AI086025-01 (2009): $498908
MICROBES IN HEALTH AND DISEASE TRAINING PROGRAM
Bruce Steven Klein, Professor
University Of Wisconsin Madison, 21 N. Park Street, Suite 6401, Madison, Wi 53715-1218
Grant 5T32AI055397-08 from National Institute Of Allergy And Infectious Diseases
Abstract: A community of investigators seeks renewed support for a pre- and postdoctoral training program that addresses the role of microbes in health and disease at the University of Wisconsin. Microbiology is fundamentally important to human health due to the prevalence and consequences of infectious diseases. Its significance has been elevated by bioterrorism and discoveries of the unforseen roles for microbes in certain human maladies and in promoting normal human physiology and health. The proposed Microbes in Health and Disease training program will have its physical and intellectual home in a new Microbial Sciences Building where basic and clinical scientists interact and collaborate, providing a strong sense of place, cohesion and identity to the Training Program. Our predoctoral trainees are drawn from the Microbiology Doctoral Training Program (MDTP), a highly ranked graduate program. Our postdoctoral fellows are drawn from a strong pool of PhD and Infectious Disease MD fellows, the latter from a program with a long history of placing fellows into academic medicine.. A program-wide Microbial Pathogenesis and Host Response Group (MPHRG) and Symbiosis Group hold biweekly meetings together, host invited speakers and have a Website and listserver. Medical Microbiology and Immunology, Bacteriology, Internal Medicine, and Pediatrics are core departments of MDTP, MPHRG, and Symbiosis activities, and offer required didactic, journal club and seminar courses to our trainees. Instruction is provided in host-microbe interactions, microbial pathogenesis, immunology and infectious disease, translational medicine, and the responsible conduct of research. Our 28 faculty trainers span 11 departments in 4 colleges and actively collaborate with each other in both research and teaching. All trainers are productive scientists with proven NIH or equivalent funding records and strong records of graduate training. Most are tenured (15 full, 6 associate professors) and promising junior faculty trainers will be mentored by senior training faculty. The training program faculty are dedicated to recruiting outstanding students and fellows, including focused efforts for minority candidates, and are committed to pre- and postdoctoral mentoring and didactic and research training. To support this committment, and the NIH-stated need to train scientists in the area of microbes in health and disease, a request is made for the support of 8 trainees annually 5 predoctoral trainees, and 3 postdoctoral trainees, including PhD and MD fellows. Each trainee is mentored by a committee consisting of a thesis advisor or mentor and 4 other faculty, and all trainees are also co-mentored by virtue of joint trainer service on these committees. The program and its trainers are highly regarded in the scientific community, and the success of the training program in the past cycle is demonstrated by the collective 29 publications of the 14 trainees, and their progress into competitive postdoctoral positions or academic, industry or government research careers
Keywords: Disease; Disorder; Health; Microbe; Training Programs; disease/disorder
Project start date: 2003-08-01
Project end date: 2013-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
PFA/PA: PA-06-468
5T32AI055397-08 (2010): $183325
5T32AI055397-07 (2009): $165465
Molecular Pathogenesis Of Blastomycosis
Bruce Steven Klein, Professor
Pediatricsuniversity Of Wisconsin Madison
Grant 4R37AI035681-15 from National Institute Of Allergy And Infectious Diseases IRG: NSS
Project start date: 1995-01-01
Project end date: 2013-12-31