Ocular HSV-1: Turning On/Off LAT Expression In Latency
Guey-chuen Perng
Cedars-sinai Medical Center
los Angeles, Ca 90048
Grant 1R03EY013701-01 from National Eye Institute IRG: ZEY1
Abstract: Applicant´s ) Recurrent herpes simplex virus type I (HSV-1) infection is a major cause of viral induced blindness. Although the HSV-1 LAT gene is essential for efficient spontaneous reactivation, it remains unclear whether LAT´s main latency related function takes place during establishment of latency or reactivation from latency. Although LAT can enhance the rate at which latency is established by 2-3 fold, it can enhances spontaneous reactivation by up to 10-fold. In addition, a mutation in a gene other than LAT has been shown to reduce establishment of latency by 20-fold without altering spontaneous reactivation. We hypothesize that LAT has an important function during the reactivation stage of the HSV-1 latency reactivation cycle. This would make LAT an excellent target for therapeutic intervention to block recurrent disease, and would have tremendous clinical significance. Our specific aim is to construct and then infect rabbits with an HSV-1 mutant in which LAT is expressed only when Dox is present.in the rabbit´s water supply. Dox will be fed either (i)just prior to and then throughout the first 21 days of infection (LAT will be active only during the time of establishment of latency) or (ii)continuously from days 22 to 60 post infection (pi) (LAT will be active only during the time of reactivation from latency). Spontaneous reactivation will be determined from 30 to 60 days pi. We expect these two complementary experiments to show that LAT is important during reactivation. Thus, (i)when LAT is active only during establishment we expect spontaneous reactivation to be reduced to LAT null mutants levels; and (10when LAT is active only during reactivation we expect wild type spontaneous reactivation levels. These results would confirm that LAT is important at the reactivation step and suggest that blocking LAT´s function would be an outstanding therapeutic approach for eliminating or reducing recurrent HSV-1 ocular disease in individuals with a history of recurrent ocular HSV-1
Keywords: gene expression, gene induction /repression, herpes simplex virus 1, latent virus infection, ocular herpes, relapse /recurrence, virus infection mechanism gene mutation laboratory rabbit, molecular cloning, northern blotting, regulatory gene, southern blotting, transgenic animal
Project start date: 2001-09-01
Project end date: 2002-06-30
1R03EY013701-01 (2001): $153000
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Guey-chuen Perng
EARLY INNATE IMMUNE RESPONSE IN DENGUE VIRUS INFECTION
Guey-chuen Perng, Dr.
Emory University, 1599 Clifton Road, 4th Floor, Atlanta, Ga 30322
Abstract: This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The objective of this study has been to document the early dissemination and pathology of DENGUE virus infection in rhesus macaques. Human infection with Dengue virus is generally detected about one week post infection and there is therefore a dearth of information on the early event of virus infection in a model close to man. Infection of 6 rhesus macaque with a high dose of Dengue serotype 2 intravenously has for the first time induced reproducible albeit generally mild coagulopathies starting on day 3 and lasting to 7-10 days post infection. These coagulopathies have resulted in subcutaneous widely distributed ecchymoses as well as generally disseminated petechia in the most severe cases. The mechanisms in the coagulation cascade that are impaired by Dengue infection in vivo appear to center on protein C and may provide us a novel therapeutic window for fatal human dengue hemorrhagic fever in humans. These studies have allowed us to define the initial targets of Dengue virus which appear to target megakaryocytes and platelets. Finally the role of innate responses and dendritic cells and macrophage in particular can be evaluated during the acute phase of infection in vivo using this model
Keywords: Acute; Bizzozero`s corpuscle/cell; Blood Coagulation Disorders; Blood Platelets; Blood megakaryocyte; Breakbone Fever Virus; CRISP; Clotting; Coagulation; Coagulation Disorder; Coagulation Process; Coagulopathy; Computer Retrieval of Information on Scientific Projects Database; Deetjeen`s body; Dendritic Cells; Dengue; Dengue Fever; Dengue Hemorrhagic Fever; Dengue Virus; Dose; Ecchymosis; Event; Funding; Grant; Hayem`s elementary corpuscle; Human; Human, General; Immune response; Infection; Institution; Investigators; Macaca mulatta; Mammals, Primates; Man (Taxonomy); Man, Modern; Marrow platelet; Megakaryocytes; Megalokaryocyte; Modeling; NIH; National Institutes of Health; National Institutes of Health (U.S.); Pathology; Petechiae; Phase; Platelets; Primates; Protein C; Research; Research Personnel; Research Resources; Researchers; Resources; Reticuloendothelial System, Platelets; Rhesus; Rhesus Macaque; Rhesus Monkey; Role; Serotyping; Source; Thrombocytes; Time; United States National Institutes of Health; Veiled Cells; Viral Diseases; Virus Diseases; breakbone fever; clotting disorder; host response; immunoresponse; in vivo; macrophage; man; man`s; new therapeutics; next generation therapeutics; novel therapeutics; response; social role; subcutaneous; thrombocyte/platelet; viral infection; virus infection
Project start date: 2009-05-01
Project end date: 2010-04-30
Budget start date: 1-MAY-2009
Budget end date: 30-APR-2010
5P51RR000165-49_8426 (2009): $43907
NEUROLOGICAL DISORDER: ROLE OF THE UOL IN CNS IMMUNE SYSTEM
Guey-chuen Perng, Dr.
Emory University, 1599 Clifton Road, 4th Floor, Atlanta, Ga 30322
Grant 5R01NS049556-04 from National Institute Of Neurological Disorders And Stroke
Abstract: Herpes simplex virus (HSV) infects the eyes, replicates and travels through axons to the sensory neurons, where it becomes latent. Sporadically the virus can reactivate, return to the eye and cause corneal scarring and loss of vision. We have cloned a new HSV-1 gene, designated UOL, that is located in a genomic region previously suggested to be involved in viral virulence. Based upon two-hybrid system preliminary studies, UOL protein interacts strongly with ICAM-5 (intercellular adhesion molecules 5) from mouse brain. ICAM-5 is an important molecule in neuron-leukocyte interactions that is proposed to attract immune cells to the CMS. In pull-down experiments using lysates from mouse brains, UOL is able to pull-down ICAM-5. In addition, delayed immune response in immune lymphocytes infiltrating to CNS in wild type compared to that of UOL deletion virus infected mice. Thus, we hypothesize that UOL-ICAM-5 interactions play an important role in HSV virulence. To test this hypothesis, we propose the following specific aims (1) Confirm that UOL-protein binds ICAM-5 in vitro and in vivo by protein-protein pull-down assays, co-immunoprecipitation, and confocal microscopy. (2)Confirm that UOL-ICAM-5 interactions are involved in HSV-1 neurovirulence. Several viral mutants will be constructed including anti-sense ICAM-5 oligonucleotides that can block ICAM-5 expression, UOL-shut off, and UOL-turn on mutants. By blocking ICAM-5 will significantly increase virulence of the HSV UOL mutant, while over express UOL should produce increased virulence. (3) Dissect the effects of immune cells by the interaction of UOL with ICAM-5 in CNS. ICAM-5 regulates immune response in CNS on the setting of infection. Immune lymphocytes infiltrating into CNS will be characterized and cytokines profile will be documented. Taken together, theses aims will shed new light on how HSV evades CNS immune system and may lead to the development of new treatments for HSV encephalitis
Keywords: (Z)-2-[4(1, 2-diphenyl-1-butenyl)-phenoxyl]-N, N-dimethylethanamine; 1-p-beta-dimethylamino-ethoxyphenyl-trans-1, 2-diphenylbut-1-ene; 2-Naphthacenecarboxamide, 4-(dimethylamino)-1, 4, 4a, 5, 5a, 6, 11, 12a-octahydro-3, 5, 10, 12, 12a-pentahydroxy-6-methyl-1, 11-dioxo-, (4S-(4alpha, 4aalpha, 5alpha, 5aalpha, 6alpha, 12aalpha))-; 21+ years old; Adhesives; Adult; Affect; Afferent Neurons; Animals; Apoptosis; Apoptosis Pathway; Assay; Axon; Bioassay; Biologic Assays; Biological Assay; Blindness; Blood leukocyte; Blotting, Western; Brain; Cell Communication; Cell Death, Programmed; Cell Interaction; Cell-to-Cell Interaction; Cells; Cerebrum; Cessation of life; Chimera Protein; Chimeric Proteins; Clinical; Co-Immunoprecipitations; Complex; Confocal Microscopy; Data; Death; Development; Doxycycline; Encephalitis; Encephalon; Encephalons; Ethanamine, 2-(4-(1, 2-diphenyl-1-butenyl)phenoxy)-N, N-dimethyl-, (Z)-; Eye; Eyeball; Fusion Protein; Gasser`s Ganglion; Gasserian Ganglion; Genes; Genomics; Goals; HHV-1; HSV; HSV-1; HSV1; Herpes Simplex Virus; Herpes Simplex Virus 1; Herpes Simplex Virus Type 1; Herpes labialis Virus; Herpesvirus 1 (alpha), Human; Herpesvirus 1, Human; Herpesvirus hominis; Human herpes simplex virus type 1; Human herpesvirus 1; Human herpesvirus type 1; Human, Adult; ICAM-5; Immune; Immune response; Immune system; In Vitro; Incidence; Infection; Inflammation, Brain; Intercellular Adhesion Molecules; Investigators; Lead; Lesion; Leukocytes; Light; Location; Lymphocyte; Lymphocytic; Mammals, Mice; Marrow leukocyte; Mice; Moab, Clinical Treatment; Molecular; Monoclonal Antibodies; Murine; Mus; Nerve Cells; Nerve Unit; Nervous System Diseases; Nervous System, Brain; Neural Cell; Neurocyte; Neurologic Disorders; Neurological Disorders; Neurons; Neurons, Afferent; Neurons, Sensory; Nucleases, RNA; Oligo; Oligonucleotides; Pathogenesis; Pb element; Photoradiation; Play; Programs (PT); Programs [Publication Type]; Protein Binding; Proteins; Publications; RNase; Recurrence; Recurrent; Reporting; Research Personnel; Researchers; Reticuloendothelial System, Leukocytes; Ribonuclease Family Protein; Ribonucleases; Role; Scientific Publication; Semilunar Ganglion; Sensory Cell Afferent Neuron; Serologic; Serological; Simplexvirus; Staining method; Stainings; Stains; Stress; Structure of trigeminal ganglion; Survival Rate; Symptoms; System; System, LOINC Axis 4; TAM; Tamoxifen; Testing; Thick; Thickness; Time; Transcript; Transgenes; Transgenic Organisms; Travel; Trigeminal Ganglias; Trigeminal Ganglion; Two Hybrid; Vibramycin; Viral; Virulence; Virus; Virus Replication; Viruses, General; Western Blotting; Western Blottings; Western Immunoblotting; White Blood Cells; White Cell; Yeast One Hybrid System; Yeast One/Two-Hybrid System; adult human (21+); alpha-6-Deoxyoxytetracycline; base; body system, allergic/immunologic; cDNA Library; cell type; cicatrix corneal; corneal scar; cryostat; cytokine; experiment; experimental research; experimental study; gene product; heavy metal Pb; heavy metal lead; herpes simplex i; herpes virus 1, human; herpesvirus; host response; human alphaherpesvirus 1; human herpesvirus 1 group; immunoresponse; in vivo; latency associated transcript; lymph cell; mutant; nervous system disorder; neurological disease; neuronal; neurotropic; neurovirulence; organ system, allergic/immunologic; overexpression; programs; protein blotting; research study; response; social role; telencephalin; transgenic; virus multiplication; white blood cell; white blood corpuscle; yeast two hybrid system
Project start date: 2007-09-30
Project end date: 2011-06-30
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5R01NS049556-04 (2010): $306727
5R01NS049556-03 (2009): $309825
5R01NS049556-02 (2008): $309825
1R01NS049556-01A1 (2007): $330075
Ocular HSV-1: Turning On/Off LAT Expression In Latency
Guey-chuen Perng
University Of California Irvine Irvine, Ca 926977600
Grant 5R03EY013701-04 from National Eye Institute IRG: ZEY1
Abstract: Applicant s ) Recurrent herpes simplex virus type I (HSV-1) infection is a major cause of viral induced blindness. Although the HSV-1 LAT gene is essential for efficient spontaneous reactivation, it remains unclear whether LAT s main latency related function takes place during establishment of latency or reactivation from latency. Although LAT can enhance the rate at which latency is established by 2-3 fold, it can enhances spontaneous reactivation by up to 10-fold. In addition, a mutation in a gene other than LAT has been shown to reduce establishment of latency by 20-fold without altering spontaneous reactivation. We hypothesize that LAT has an important function during the reactivation stage of the HSV-1 latency reactivation cycle. This would make LAT an excellent target for therapeutic intervention to block recurrent disease, and would have tremendous clinical significance. Our specific aim is to construct and then infect rabbits with an HSV-1 mutant in which LAT is expressed only when Dox is present.in the rabbit s water supply. Dox will be fed either (i)just prior to and then throughout the first 21 days of infection (LAT will be active only during the time of establishment of latency) or (ii)continuously from days 22 to 60 post infection (pi) (LAT will be active only during the time of reactivation from latency). Spontaneous reactivation will be determined from 30 to 60 days pi. We expect these two complementary experiments to show that LAT is important during reactivation. Thus, (i)when LAT is active only during establishment we expect spontaneous reactivation to be reduced to LAT null mutants levels; and (10when LAT is active only during reactivation we expect wild type spontaneous reactivation levels. These results would confirm that LAT is important at the reactivation step and suggest that blocking LAT s function would be an outstanding therapeutic approach for eliminating or reducing recurrent HSV-1 ocular disease in individuals with a history of recurrent ocular HSV-1.
Keywords: gene expression, gene induction /repression, herpes simplex virus 1, latent virus infection, ocular herpes, relapse /recurrence, virus infection mechanism, gene mutation, laboratory rabbit, molecular cloning, northern blotting, regulatory gene, southern blotting, transgenic animal
Project start date: 2001-09-01
Project end date: 2004-08-31
5R03EY013701-04 (2003): $151500
5R03EY013701-03 (2002): $151500