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Genome-wide analysis of CD8 cells laser captured from skin biopsies during the mucocutaneous HSV-2 reactivation

ABSTRACT: Analysis of CD8 cells at the gene expression level during the genital HSV-2 reactivation. We would like to get a comprehensive overview on gene expression of CD8 T-cells during the post healing phases of HSV-2 reactivation. The hypothesis is that CD8 T-cells play a immune surveillance function during the post healing phases.

ORGANISM(S): Homo sapiens

PROVIDER: GSE39625 | GEO | 2012/07/26

SECONDARY ACCESSION(S): PRJNA171314

REPOSITORIES: GEO

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Genome-wide analysis of CD8 cells laser captured from skin biopsies during the mucocutaneous HSV-2 reactivation

Project description:Analysis of CD8 cells at the gene expression level during the genital HSV-2 reactivation. We would like to get a comprehensive overview on gene expression of CD8 T-cells during the post healing phases of HSV-2 reactivation. The hypothesis is that CD8 T-cells play a immune surveillance function during the post healing phases. CD8 cells were immunofluorescently stained and laser captured from post healing biopsies and contralateral control biopsies, total RNA were purified from captured cells and cDNA were amplified from the total RNA using NuGEN pico kits

2012-07-25 | E-GEOD-39625 | biostudies-arrayexpress

Genome-wide analysis of keratinocytes laser captured from skin biopsies during the mucocutaneous HSV-2 reactivation

Project description:We aim to analyze the transcriptional profiles of keratinocytes near dermal-epidemal junction during HSV-2 reactivation in humans, including lesion, post healed (4 and 8 weeks post healing) and contra-lateral control biopsies. Since keratinocytes are the main target cells for HSV-2 infection, the goal is to define the intrinsic immunity of keratinocytes during HSV-2 reactivation in humans.

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Genome-wide analysis of skin biopsies during the mucocutaneous HSV-2 reactivation

Project description:Analysis of host response at the gene expression level during the genital HSV-2 reactivation. We would like to get a comprehensive overview on host gene expression and correlate them with the infiltrated immune cell types during HSV-2 reactivation. The hypothesis is that HSV-2 may evade some aspects of host immune response during the recurrent infection.

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Transcriptional profiling of CD8 cells laser captured from skin biopsies at dermal-epidermal junction and at deeper dermal area near blood vessel during the healed phase of mucocutaneous HSV-2 reactivation

Project description:We aim to get a comprehensive analysis on gene expression of CD8a+ T-cells during the post healed phases of HSV-2 reactivation at different anatomic location (CD8a+ cells at dermal-epidermal junction versus CD8a+ cells in deeper dermal area near blood vessel). Our hypothesis is that JC_CD8 cells may function as a resident effector-like T-cells while BV_CD8 cells may be infiltrating cells from circulation. CD8 cells were immunofluorescently stained and laser captured from genital skin biopsies at dermal-epidermal junction at 2 and 8 weeks post healed (JC_2wks_CD8 and JC_8wks_CD8) and at deeper dermal area near blood vessel (BV_2wks_CD8 and BV_8wks_CD8); total RNA were purified from captured cells and cDNA were amplified from the total RNA using NuGEN pico kits

2013-04-11 | E-GEOD-44975 | biostudies-arrayexpress

Transcriptional profiling of human genital skin biopsies at lesion, 2- and 8-weeks post healing and contra-lateral control genital skin biopsies during recurrent HSV-2 infection

Project description:Analysis of host response at the gene expression level during lesion and post healing periods (2- & 8-weeks after HSV-2 reactivation induced lesion is resolved). We would like to get a comprehensive overview on host gene expression of genital skin during lesion and post healing periods of recurrent HSV-2 infection as compared to those in control skin.

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2013-04-11 | GSE44975 | GEO

Therapeutic Prime/Pull Vaccination of HSV-2 Infected Guinea Pigs with the Ribonucleotide Reductase 2 (RR2) and CXCL11 boosts TRM and TEM CD4+/CD8+ T cells to protect from recurrent genital herpes

Project description:Following acute herpes simplex virus type 2 (HSV-2) infection, the virus undergoes an asymptomatic latent infection of sensory neurons of dorsal root ganglia (DRG). Chemical and physical stress cause intermittent virus reactivation from latently infected DRG and recurrent virus shedding in the genital mucosal epithelium causing genital herpes in symptomatic patients. While T-cells appear to play a role in controlling virus reactivation from DRG and reducing the severity of recurrent genital herpes, the mechanisms for recruiting these T-cells into DRG and the vaginal mucosa (VM) remain to be fully elucidated. The present study investigates the effect of CXCL9, CXCL10, and CXCL11 T-cell-attracting chemokines on the frequency and function of DRG- and VM-resident CD4+ and CD8+ T cells and its effect on the frequency and severity of recurrent genital herpes in the recurrent herpes guinea pig model. HSV-2 latent-infected guinea pigs were immunized intramuscularly with the HSV-2 RR2 protein (Prime) and subsequently treated intravaginally with the neurotropic adeno-associated virus type 8 (AAV-8) expressing CXCL9, CXCL10, or CXCL11 chemokines to recruit CD4+ and CD8+ T cells into the infected DRG and VM (Pull). Compared to the RR2 therapeutic vaccine alone, the RR2/CXCL11 prime/pull therapeutic vaccine significantly increased the frequencies of functional tissue-resident (TRM cells) and effector (TEM cells) memory CD4+ and CD8+ T cells in both DRG and VM tissues. This was associated with less virus in the healed genital mucosal epithelium and reduced frequency and severity of recurrent genital herpes. These findings confirm the role of local DRG- and VM-resident CD4+ and CD8+ T cells in reducing virus shedding at the vaginal site of infection and the severity of recurrent genital herpes and propose the novel prime-pull vaccine strategy to protect against recurrent genital herpes.

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Effect of NK1.1+ cell depletion on vaginal neutrophil gene expression during HSV-1 infection

Project description:Depletion of NK1.1+ cells from mice prior to intravaginal HSV-1 infection results in severely increased genital disease and mucosal tissue damage. We determined that this loss of host protection in the absence of NK1.1+ cells was dependent on neutrophils, suggesting that NK1.1+ cells may be modulating neutrophil behavior during infection. To obtain an unbiased view of the transcriptional changes occuring in neutrophils that might explain this difference in disease outcomes, we performed sequencing of total RNA from neutrophils isolated from vaginas of intact or NK1.1 depleted mice 5 days after HSV-1 infection.

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Transcriptome analysis of HSV-1 infected dorsal root ganglia and large intestinal musculara

Project description:Following genital HSV-1 infection in mice, virus spreads to and infects the sensory neurons of the dorsal root ganglia (DRG) and the enteric neurons situated within the large intestinal musculara (LIM). We used RNA sequencing to simultaneously study the host and viral transcriptomes at these two distinct sites of infection. We find that transcription across the HSV-1 genome is weak and incomplete in sensory neurons whereas all known HSV-1 coding genes are robustly expressed in the enteric neurons. Furthermore, a T cell activation signature dominated the immune response to HSV-1 infection in the DRG. In contrast, an innate inflammatory signature was observed in the LIM.

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Human cellular models for Herpes Simplex Virus, type 1 (HSV-1) using induced pluripotent stem cell derived neuronal lineages

Project description:Herpes simplex virus type 1 (HSV-1) is a 152 Kb double stranded DNA alpha-herpesvirus, which establishes long life latent infection in sensory neurons. Most of our knowledge regarding HSV-1 latency comes from in vivo studies using small animal models, mainly rodents and rabbits, which are not naturally infected by HSV-1. Furthermore, these animal models do not fully recapitulate the species specific effects of human HSV-1 infection. Human cellular models utilize trigeminal ganglia removed from cadavers or, alternatively, neuron-like cells derived from cancerous cell lines that do not fully reflect effects on normal human neurons. This limitation poses the need to develop an in vitro model to investigate molecular details of the mechanisms underlying latency and reactivation in human neurons. Induced pluripotent stem (iPS) cell technologies offer an unprecedented opportunity to generate unlimited supplies of neurons and the facility to manipulate such cells in vitro. In this study, we developed an in vitro HSV-1 infection model in human iPS-derived neural progenitor cells (NPCs) and neurons, which displays the main hallmarks of latency defined in animal models and in humans. Induced pluripotent stem (iPS) cells were generated from human skin biopsy samples

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