Recurrent herpes simplex virus type 1 (HSV-1) infection is a major cause of viral induced blindness. Elucidation of the underlying molecular mechanisms behind the HSV-1 latency-reactivation cycle, should lead to development of a means for reducing the incidence of HSV-1 induced blindness. The HSV-1 LAT gene is essential for the high wild type reactivation phenotype and this appears to be related to LAT's antiapoptosis activity. We plan to confirm that LAT functions as an RNA, that LAT's ability to block multiple apoptosis pathways is critical for its ability to enhance reactivation, and that LAT plays an important role at the time of reactivation.
Our specific aims are: 1. Confirm the hypothesis that LAT blocks apoptosis via its RNA rather than via a LAT protein. We will: a) Knock out all 4 potential LAT ORFs within the functional region of LAT and confirm that these plasmids still efficiently block apoptosis and that introducing these knock outs into the virus do not reduce the reactivation phenotype; b) Confirm that expression of the LAT ORFs (each in a separate plasmid) do not block apoptosis, either individually or together; c) Show that LAT can block apoptosis even when protein synthesis is blocked. 2. Confirm the hypothesis that LAT enhances the reactivation phenotype by blocking apoptosis at multiple steps and/or pathways. We will replace LAT in the virus with alternative anti-apoptosis genes that block specific apoptotic pathways. We predict that only alternative anti-apoptosis genes that block multiple apoptosis steps will be able to substitute for LAT and restore the high reactivation phenotype. We will also investigate the mechanism by which LAT RNA blocks apoptosis. 3. Test the hypothesis that LAT's anti-apoptotic activity plays an important role at the time of reactivation. HSV-1 mutants will be engineered such that LAT expression will be turned on or off in specific transgenic mice following treatment with doxycycline or Tamoxifen. The reactivation phenotype of these mutant viruses will be determined when LAT is expressed only during establishment of latency compared to when LAT is expressed only during reactivation from latency.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY013191-06
Application #
7099512
Study Section
Special Emphasis Panel (ZRG1-AED (01))
Program Officer
Shen, Grace L
Project Start
2000-07-01
Project End
2009-05-31
Budget Start
2006-06-01
Budget End
2007-05-31
Support Year
6
Fiscal Year
2006
Total Cost
$489,087
Indirect Cost
Name
University of California Irvine
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Perng, Guey-Chuen; Osorio, Nelson; Jiang, Xianzhi et al. (2016) Large Amounts of Reactivated Virus in Tears Precedes Recurrent Herpes Stromal Keratitis in Stressed Rabbits Latently Infected with Herpes Simplex Virus. Curr Eye Res 41:284-91
Jiang, Xianzhi; Brown, Don; Osorio, Nelson et al. (2016) Increased neurovirulence and reactivation of the herpes simplex virus type 1 latency-associated transcript (LAT)-negative mutant dLAT2903 with a disrupted LAT miR-H2. J Neurovirol 22:38-49
BenMohamed, Lbachir; Osorio, Nelson; Khan, Arif A et al. (2016) Prior Corneal Scarification and Injection of Immune Serum are Not Required Before Ocular HSV-1 Infection for UV-B-Induced Virus Reactivation and Recurrent Herpetic Corneal Disease in Latently Infected Mice. Curr Eye Res 41:747-56
Srivastava, Ruchi; Dervillez, Xavier; Khan, Arif A et al. (2016) The Herpes Simplex Virus Latency-Associated Transcript Gene Is Associated with a Broader Repertoire of Virus-Specific Exhausted CD8+ T Cells Retained within the Trigeminal Ganglia of Latently Infected HLA Transgenic Rabbits. J Virol 90:3913-3928
Jester, James V; Morishige, Naoyuki; BenMohamed, Lbachir et al. (2016) Confocal Microscopic Analysis of a Rabbit Eye Model of High-Incidence Recurrent Herpes Stromal Keratitis. Cornea 35:81-8
Khan, Arif A; Srivastava, Ruchi; Spencer, Doran et al. (2015) Phenotypic and functional characterization of herpes simplex virus glycoprotein B epitope-specific effector and memory CD8+ T cells from symptomatic and asymptomatic individuals with ocular herpes. J Virol 89:3776-92
Carpenter, Dale; Hsiang, Chinhui; Jiang, Xianzhi et al. (2015) The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) protects cells against cold-shock-induced apoptosis by maintaining phosphorylation of protein kinase B (AKT). J Neurovirol 21:568-75
Srivastava, Ruchi; Khan, Arif A; Spencer, Doran et al. (2015) HLA-A02:01-restricted epitopes identified from the herpes simplex virus tegument protein VP11/12 preferentially recall polyfunctional effector memory CD8+ T cells from seropositive asymptomatic individuals and protect humanized HLA-A*02:01 transgenic mice J Immunol 194:2232-48
Jiang, Xianzhi; Brown, Don; Osorio, Nelson et al. (2015) A herpes simplex virus type 1 mutant disrupted for microRNA H2 with increased neurovirulence and rate of reactivation. J Neurovirol 21:199-209
BenMohamed, Lbachir; Osorio, Nelson; Srivastava, Ruchi et al. (2015) Decreased reactivation of a herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) mutant using the in vivo mouse UV-B model of induced reactivation. J Neurovirol 21:508-17

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