David M Underhill
Cedars-sinai Medical Center
Project start date: 2008-01-01
Project end date: 2012-12-31
Sponsored Links Excellgen http://Excellgen.com
NON-TOLL-LIKE RECEPTOR INNATE IMMUNE SIGNALING
David M Underhill, Associate Professor
Cedars-sinai Medical Center, Los Angeles, Ca 90048
Grant 5R01AI071116-03 from National Institute Of Allergy And Infectious Diseases
Keywords: ARAM; Active Oxygen; Affect; Ag Recognition Activation Motif; Agonist; Animal Welfare; Antigen Receptors; Assay; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Bibliography; Binding; Binding (Molecular Function); Bioassay; Biologic Assays; Biological Assay; Blood Neutrophil; Blood Polymorphonuclear Neutrophil; Blood Segmented Neutrophil; Blood Serum; CSIF; CSIF-10; Caring; Cell Communication and Signaling; Cell Function; Cell Process; Cell Signaling; Cell physiology; Cells; Cellular Function; Cellular Infiltration; Cellular Physiology; Cellular Process; Ciclosporin; Complement; Complement Proteins; Complex; Country; CsA; Cyclosporin A; Cyclosporine; Cyclosporine A; Cytokine Gene; Cytokine Receptors; Cytokine Synthesis Inhibitory Factor; Cytokine formation-inhibiting factor (mouse clone F115 protein moiety reduced); Cytokines, Chemotactic; Data; Defect; Dendritic Cells; Development; Differentiation Factor, B-Cell; EC 2.7; Ecological impact; Edodekin Alfa; Environment; Environmental Impact; Equipment; Ethics Committees, Research; Exposure to; Family; Genes; Genes, c-fms; Glucans; Glucose Polymer; HPGF; Hepatocyte-Stimulating Factor; Heterophil Granulocyte; Homologous Chemotactic Cytokines; Host Defense; Hybridoma Growth Factor; IACUC; IFN-beta 2; IFNB2; IL-10; IL-12; IL-6; IL10; IL10A; IL12; IL6 Protein; INFLM; IRBs; ITAM; Immune; Immune response; Immunologic Receptors; Immunological Receptors; Immunoreceptor Tyr-Based Activation Motif; Immunoreceptor Tyrosine-Based Activation Motif; Impact, Environmental; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Institutional Animal Care and Use Committee; Institutional Review Boards; Integral Membrane Protein; Intercrines; Interleukin 10 Precursor; Interleukin 6 (Interferon, Beta 2); Interleukin-10; Interleukin-12; Interleukin-6; International; Intracellular Communication and Signaling; Intrinsic Membrane Protein; Killings; Kinases; Knock-out; Knockout; MGI-2; Marrow Neutrophil; Mediating; Methods; Mice, Transgenic; Modeling; Molecular Interaction; Myeloid Differentiation-Inducing Protein; NF-AT proteins; NFAT proteins; NFATC proteins; NKSF; Natural Killer Cell Stimulatory Factor; Neutrophilic Granulocyte; Neutrophilic Leukocyte; Oxygen Radicals; Pathway interactions; Peptides; Peritoneal; Phagocytes; Phagocytic Cell; Phagocytosis; Phosphotransferases; Plasmacytoma Growth Factor; Play; Polyglucoses; Polymorph; Polymorphonuclear Cell; Polymorphonuclear Leukocytes; Polymorphonuclear Neutrophils; Principal Investigator; Pro-Oxidants; Production; Programs (PT); Programs [Publication Type]; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Protein-Tyrosine Kinases, src; Reactive Oxygen Species; Receptor Protein; Receptor Signaling; Receptors, Antigen; Receptors, Cytokine; Receptors, Immunologic; Relative; Relative (related person); Research; Research Ethics Committees; Research Resources; Resources; Role; SIS cytokines; Sandimmun; SangCya; Serum; Shapes; Side; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Sound; Sound - physical agent; Stimulus; Subcellular Process; Swelling; T cell regulation; T-Cell Activation; TIL4; TLR protein; TLR2; TLR2 gene; Tail; Toll-like receptors; Toll/Interleukin 1 Receptor-Like 4 Gene; Transgenic Mice; Transgenic Organisms; Translating; Translatings; Transmembrane Protein; Transphosphorylases; Uncertainty; Veiled Cells; Vertebrate Animals; Vertebrates; Yeasts; Zymosan; ing; acquired immunity; amebocyte; anti-microbial; antimicrobial; base; biological signal transduction; c fms; c-fms Proto-Oncogenes; chemoattractant cytokine; chemokine; clinical significance; clinically significant; cytokine; dectin 1; doubt; experiment; experimental research; experimental study; expiration; gene induction; host response; human subject; immune receptor; immunoresponse; in vivo; interest; interferon beta 2; language translation; macrophage; microbial; mouse model; neoral; neutrophil; nuclear factor of activated T-cells, cytoplasmic; nuclear factors of activated T-cells; particle; pathogen; pathway; programs; receptor; research study; response; sandimmune; social role; sound; src Kinases; src Tyrosine Kinases; src-Family Kinases; src-Family Tyrosine Kinases; transcription factor NF-AT; transgenic; vertebrata
Project start date: 2008-01-01
Project end date: 2012-12-31
Budget start date: 1-JAN-2010
Budget end date: 31-DEC-2010
PFA/PA: PA-07-070
5R01AI071116-03 (2010): $357750
Grants awarded to David M Underhill
ANALYSIS OF TOLL LIKE RECEPTOR SIGNALING COMPLEXES
David M Underhill, Assistant Professor
Scripps Research Institute La Jolla, Ca 920371000
Grant 1U54AI054523-010007 from National Institute Of Allergy And Infectious Diseases IRG: ZGM1
Abstract: The purpose of this bridging grant is to generate a complete register of all the signaling molecules associated with Toll-like receptors (TLRs) before and after stimulation with ligands, and a full of the downstream signaling complexes that form within the cytoplasm of the activated cell. Only with a complete understanding of the molecular switching mechanisms controlling inflammatory signaling can we intelligently identify molecules that may control susceptibility to disease or prove to be important candidates for therapeutic intervention. Ongoing projects supported in the lab include the analysis of Toll-like receptors domains that mediate signal activation and association with known signaling molecules such as MyD88 and IRAK. While the importance of these molecules in macrophage activation by pathogens is clear, it is also clear that we have not yet identified many more equally important proteins that function in signaling initiated by TLRs. Present constraints on project definition and scope require individual laboratories to study TLR signaling by standard incremental hypothesis-driven approaches, while the resources and collaborative environment of the GLUE consortium make the full, unbiased analysis of the TLR-initiated signaling cascade possible. By identifying the full set of proteins that mediate LPS-induced (TLR4-mediated) signaling in macrophages, and mapping the organization structure of these components, we will produce a novel foundation for further studies on the regulation of inflammatory responses during sepsis.
Keywords: bacterial disease, bactericidal immunity, biological signal transduction, toll like receptor, CD14 molecule, cooperative study, inflammation, leukocyte activation /transformation, lipopolysaccharide, macrophage, protein protein interaction, protein sequence, receptor expression, electrospray ionization mass spectrometry, epitope mapping, immunoprecipitation, mass spectrometry, yeast two hybrid system
Project start date: 2002-09-30
Project end date: 2007-07-31
PHAGOSOMAL TARGETING OF CARD PROTEINS
David M Underhill, Associate Professor
Cedars-sinai Medical Center, Los Angeles, Ca 90048
Grant 3R01GM085796-03S1 from National Institute Of General Medical Sciences
Abstract: Phagosomal targeting of CARD proteins Phagocytosis is the process by which cells such as macrophages and dendritic cells bind, internalize, and kill microbes. Several different types of receptors are known to recognize microbes (either directly or through opsonization) and to trigger phagocytosis. The cell biological mechanisms by which these receptors drive phagocytosis are dependent the specific receptor. Thus, Fc-receptor mediated phagocytosis recruits a distinct set of cytoskeletal components than Dectin-1 (beta-glucan receptor) or complement receptor-mediated phagocytosis. Phagocytosis is tightly coupled to the initiation of inflammatory cytokine and chemokine production, although the mechanisms by which these processes are coupled are still being elucidated. Caspase Activation and Recruitment Domains (CARDs) are conserved protein-protein interaction domains found in a variety of cytoplasmic proteins key to microbial recognition and inflammatory signaling including Nod (nuclear oligomerization domain) proteins, Bcl10, Nalp1 and many others. We have observed that several CARD proteins are recruited to phagosomes. We hypothesize that CARD recruitment to phagosomes is a key mechanism by which signals for phagocytosis and inflammation are integrated. In this study, we will define the mechanism(s) by which CARD proteins are recruited to phagosomes and whether CARD proteins are recruited only to specific types of phagosomes (Aim1). We will determine what protein-protein interactions are required to get CARD proteins to phagosomes and whether these interactions are important for inflammatory signaling (Aim 2). We will determine whether activating a CARD protein (Nod2) on phagosomes alters its inflammatory signaling consequences, and whether such targeted activation influences antigen presentation and the type of adaptive immune response that is promoted. LAY SUMMARY White blood cells eat and kill infectious microbes. They also initiate inflammatory responses that are crucial for defense against infection. We are defining how the molecular signaling pathways that control the processes of eating and killing microbes are connected to the signaling pathways required to activate inflammation. Clinical manipulation of these pathways may help suppress unwanted inflammation, or stimulate effective immune defenses
Keywords: ATGN; Actinin; Actins; Adjuvant; Affect; Agonist; Antigen Presentation; Antigen Presentation Pathway; Antigen Processing and Presentation; Antigens; Autophagocytosis; Binding; Binding (Molecular Function); Biological; Blood leukocyte; Cell Communication and Signaling; Cell Signaling; Cell-Death Protease; Cells; Cellular Matrix; Clinical; Complement Receptor; Coupled; Crohn`s disease; Crohn`s disorder; Cytokines, Chemotactic; Cytoplasmic Protein; Cytoskeletal System; Cytoskeleton; Dendritic Cells; Diathesis; Disease susceptibility; Eating; Enteritis, Granulomatous; Fc Receptor; FcR; Food Intake; Gene Proteins; Genes; Genetic Alteration; Genetic Change; Genetic defect; Genome; Homologous Chemotactic Cytokines; ICE-like protease; INFLM; Immune; Immune response; Immunologic Receptors; Immunological Receptors; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Intercrines; Intracellular Communication and Signaling; Intracellular Membranes; Killings; Lead; Leukocytes; Link; Macrophage Activation; Marrow leukocyte; Mediating; Membrane; Microbe; Molecular; Molecular Interaction; Movement; Mucosal Immunity; Mutation; Nuclear; Organelles; Pathway interactions; Pb element; Peptide Domain; Phagocytosis; Phagolysosome; Phagosomes; Predisposition gene; Process; Production; Property; Property, LOINC Axis 2; Protein Binding Domain; Protein Binding Motif; Protein Domains; Protein Gene Products; Protein-Protein Interaction Domain; Proteins; Receptor Protein; Receptors, Immunologic; Recruitment Activity; Reticuloendothelial System, Leukocytes; Role; Route; SIS cytokines; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Susceptibility Gene; Tertiary Protein Structure; Veiled Cells; White Blood Cells; White Cell; ing; antibody receptor; autophagy; beta-glucan receptor; biological signal transduction; body movement; caspase; chemoattractant cytokine; chemokine; cystein protease; cystein proteinase; cysteine endopeptidase; cytokine; dectin 1; disease/disorder proneness/risk; eleocolitis; gene product; genome mutation; granulomatous enterocolitis; heavy metal Pb; heavy metal lead; host response; immune receptor; immunogen; immunoresponse; in vivo; intracellular skeleton; liability to disease; macrophage; mannose receptor; membrane structure; microbial; novel; particle; pathway; predisposing gene; protein protein interaction; receptor; recruit; regional enteritis; social role; white blood cell; white blood corpuscle
Project start date: 2010-06-15
Project end date: 2011-05-31
Budget start date: 15-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
3R01GM085796-03S1 (2010): $48256
TOLL-LIKE RECEPTORS: ACTIVATORS OF INNATE IMMUNITY
David M Underhill, Assistant Professor
Cedars-sinai Medical Center Los Angeles, Ca 90048
Grant 7R01GM062995-07 from National Institute Of General Medical Sciences IRG: EI
Abstract: Adapted from applicant s ) The Toll-like receptors (TLRs) mediate innate immune recognition of pathogens in species as diverse as flies and humans. The purpose of this proposal is to analyze how the intracellular signaling domains of TLRs interact with each other to generate distinct pro-inflammatory signals in response to bacterial products. We demonstrated previously that while dimerization of the cytoplasmic tail of TLR4 is sufficient to induce the production of TNF-a, dimerization of the cytoplasmic tails of either TLR2 or TLR6 does not. In contrast, heterodimerization of the cytoplasmic tails of TLR2 and TLR6 does induce TNF-a. Thus, the mechanism of dimer-induced signaling is different between these two receptor pairs. We propose to map the elements of the cytoplasmic domain of TLR4 that mediate homodimer-induced signaling, and to map the elements of the cytoplasmic domains of TLR2/6 that mediate heterodimer-induced signaling. The biological consequences of these two types of signaling are different; while TLR4 homodimers induce the chemokine IP-lO, TLR2/6 heterodimers do not. We will define the regions of the cytoplasmic domain of TLR4 that specifically mediate the induction of IP- 10, and identify signaling molecules that bind to TLR4 homodimers, and not to TLR2/6 heterodimers. Although we have demonstrated previously that TLR4 mediates LPS-responses in macrophages, while TLR2/6 mediates responses to peptidoglycan, there is tantalizing evidence that under certain circumstances, TLR2 may participate in LPS-induced responses. One such circumstance is the induction of IL-12; one dominant negative mutant of TLR2 specifically ablates this response while a distinct TLR2 mutant does not. Interestingly, neither mutant blocks LPS-induced TNF-a. This distinction should permit us to map distinct areas of the cytoplasmic domain of TLR2 that participate in LPS-induced IL-12 production. We have used a rapid and robust method for mapping the signaling capacity of the cytoplasmic domains of TLR 2,4, and 6 dimers, and we will extend these studies to examine the capacity of the cytoplasmic domains of all ten TLRS to generate pro-inflammatory signals. This proposal will therefore define the signaling repertoire of different pairs of TLRs, as well as the regions of the cytoplasmic domains of these molecules responsible for triggering distinct responses such as those leading to TNF-cz, IP-lO, and IL-12.
Keywords: biological signal transduction, inflammation, microorganism immunology, receptor, CD4 molecule, IP 10 protein, chimeric protein, gene induction /repression, interleukin 12, lipopolysaccharide, macrophage, protein biosynthesis, tumor necrosis factor alpha, laboratory mouse, protein sequence, tissue /cell culture
Project start date: 2001-04-03
Project end date: 2007-03-31
7R01GM062995-07 (2005): $208547
5R01GM062995-06 (2005): $72716
5R01GM062995-05 (2004): $302861
7R01GM062995-02 (2001): $303297
David M Underhill
Cedars-sinai Medical Center
Project start date: 2008-04-01
Project end date: 2016-02-29
PHAGOSOMAL TARGETING OF CARD PROTEINS
David M Underhill, Associate Professor
Cedars-sinai Medical Center, Los Angeles, Ca 90048
Grant 5R01GM085796-03 from National Institute Of General Medical Sciences
Keywords: ATGN; Actinin; Actins; Adjuvant; Affect; Agonist; Antigen Presentation; Antigen Presentation Pathway; Antigen Processing and Presentation; Antigens; Autophagocytosis; Binding; Binding (Molecular Function); Biological; Blood leukocyte; Cell Communication and Signaling; Cell Signaling; Cell-Death Protease; Cells; Cellular Matrix; Clinical; Complement Receptor; Coupled; Crohn`s disease; Crohn`s disorder; Cytokines, Chemotactic; Cytoplasmic Protein; Cytoskeletal System; Cytoskeleton; Dendritic Cells; Diathesis; Disease susceptibility; Eating; Enteritis, Granulomatous; Fc Receptor; FcR; Food Intake; Gene Proteins; Genes; Genetic Alteration; Genetic Change; Genetic defect; Genome; Homologous Chemotactic Cytokines; ICE-like protease; INFLM; Immune; Immune response; Immunity, Mucosal; Immunologic Receptors; Immunological Receptors; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Intercrines; Intracellular Communication and Signaling; Intracellular Membranes; Killings; Lead; Leukocytes; Link; Macrophage Activation; Marrow leukocyte; Mediating; Membrane; Microbe; Molecular; Molecular Interaction; Movement; Mucosal Immunity; Mutation; Nuclear; Organelles; Pathway interactions; Pb element; Peptide Domain; Phagocytosis; Phagolysosome; Phagosomes; Predisposition gene; Process; Production; Property; Property, LOINC Axis 2; Protein Binding Domain; Protein Binding Motif; Protein Domains; Protein Gene Products; Protein-Protein Interaction Domain; Proteins; Receptor Protein; Receptors, Immunologic; Recruitment Activity; Reticuloendothelial System, Leukocytes; Role; Route; SIS cytokines; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Signaling Molecule; Susceptibility Gene; Tertiary Protein Structure; Veiled Cells; White Blood Cells; White Cell; ing; antibody receptor; autophagy; beta-glucan receptor; biological signal transduction; body movement; caspase; chemoattractant cytokine; chemokine; cystein protease; cystein proteinase; cysteine endopeptidase; cytokine; dectin 1; disease/disorder proneness/risk; eleocolitis; gene product; genome mutation; granulomatous enterocolitis; heavy metal Pb; heavy metal lead; host response; immune receptor; immunogen; immunoresponse; in vivo; intracellular skeleton; liability to disease; macrophage; mannose receptor; membrane structure; microbial; novel; particle; pathway; predisposing gene; protein protein interaction; receptor; recruit; regional enteritis; social role; white blood cell; white blood corpuscle
Project start date: 2008-04-01
Project end date: 2012-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
5R01GM085796-03 (2010): $324546