Herpes simplex virus 1 (HSV-1) infects the cornea and then establishes latency in sensory neurons of the trigeminal ganglia (TG). Sporadic spontaneous reactivation of HSV-1 causes shedding of virus in tears leading to spread of virus to other individuals, and can also cause recurrent Herpes Stromal Keratitis (HSK), a blinding ocular disease. A major gap in our current knowledge is: "How can we prevent or significantly reduce virus shedding in tears and HSV-induced ocular disease due to spontaneous reactivation of latent virus in the TG?" HSV-specific CD8+ T-cells appear to decrease in vitro induced HSV-1 reactivation in explanted mouse TG. Unfortunately, spontaneous reactivation of HSV-1 in mice is extremely rare so the relevance of these findings to in vivo HSV-1 spontaneous reactivation cannot be determined in mice. We now have a "humanized" HLA transgenic rabbit model of ocular HSV-1 that mounts "human-like" CD8 T-cell immune responses (HLA Tg rabbits). In a Preliminary Study we found that therapeutic immunization of latently infected HLA Tg rabbits with 3 human CD8 T-cell epitopes from HSV-1 gD decreased spontaneous reactivation 4-fold. This novel animal model will now allow us for the first time to test the hypothesis that a therapeutic vaccine that induces appropriate human T-cell responses to HSV-1 can decrease the effects of spontaneous reactivation (virus shedding in eyes and HSV-induced ocular disease).
Our specific Aims i nclude: (1). Test the hypothesis that therapeutic immunization with HSV-1 human CD8+ T-cell epitopes can decrease spontaneous reactivation in latently infected HLA Transgenic rabbits. CD4-CD8 lipopeptide vaccines, bearing different combinations of human CD4+ and CD8+ T cell epitopes from glycoprotein B and D (gB &gD), will be used to immunize latently infected HLA Tg rabbits. Protection against virus shedding in eyes (due to spontaneous reactivation) and HSV-induced ocular disease will be determined. (2). Test the hypothesis that the protective immunity induced by the therapeutic vaccination of HLA Transgenic rabbits in Aim 1 correlates with the presence of effector and memory CD8+ T-cells in the TG, conjunctiva, and/or draining lymph nodes. We will assess whether the number/function of HSV- and epitope- specific CD8+ T cells induced in vivo correlates with protection from spontaneous virus shedding in tears and ocular disease. We will assess the CD8+ T cell mechanism that correlates with protection. (3). Test the hypothesis that decreasing CD8+ T cells in latently infected HLA Tg rabbits will abrogate vaccine efficacy and also increase spontaneous reactivation in unvaccinated rabbits. These studies will provide important new information regarding the role of CD8+ T cells specific to human epitopes in immune control of HSV-1 spontaneous reactivation. This may lead to the development of new paradigms for immunotherapeutic strategies against ocular herpes.

Public Health Relevance

This project is aimed at developing a lipopeptide therapeutic vaccine against ocular herpes (a leading cause of blindness in developed countries) using a novel human leukocyte antigen (HLA) transgenic rabbit model.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01EY019896-05
Application #
8710229
Study Section
Vaccines Against Microbial Diseases Study Section (VMD)
Program Officer
Mckie, George Ann
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
City
Irvine
State
CA
Country
United States
Zip Code
92697
Farid, Marjan; Agrawal, Anshu; Fremgen, Daniel et al. (2016) Age-related Defects in Ocular and Nasal Mucosal Immune System and the Immunopathology of Dry Eye Disease. Ocul Immunol Inflamm 24:327-47
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
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
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
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 90:3913-28
Khan, Arif A; Srivastava, Ruchi; Chentoufi, Aziz A et al. (2015) Therapeutic immunization with a mixture of herpes simplex virus 1 glycoprotein D-derived “asymptomatic” human CD8+ T-cell epitopes decreases spontaneous ocular shedding in latently infected HLA transgenic rabbits: association with low frequency of loc J Virol 89:6619-32
Ben Haij, Nawal; Planès, Rémi; Leghmari, Kaoutar et al. (2015) HIV-1 Tat Protein Induces Production of Proinflammatory Cytokines by Human Dendritic Cells and Monocytes/Macrophages through Engagement of TLR4-MD2-CD14 Complex and Activation of NF-κB Pathway. PLoS One 10:e0129425
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
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
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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