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
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
Project description: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: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.
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.
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.
Project description:Tissue resident memory T cells (TRM) provide superior protection against infection localised to extra-lymphoid compartments in the body. Here we show that CD103+CD8+ TRM cells develop in skin from killer cell lectin-like receptor (KLR)G1-negative precursors that selectively infiltrate the epithelial layer. In the skin, a combination of chemokine-guided epithelial entry, local interleukin (IL)-15 and transforming growth factor (TGF)-β signalling is required for formation and survival of these long-lived memory cells. Importantly, TRM differentiation results in the gradual acquisition of a unique transcriptional profile that differs from that expressed by memory cells in the circulation and other types of skin-resident intra-epithelial T cells, such as the dendritic epidermal T cells (DETC). We provide a comprehensive molecular and developmental framework for the local differentiation of a distinct type of peripheral memory T cell that contributes to an important first-line of immune defence in barrier tissues such as skin and mucosa. 24 samples were analyzed: 3 replicates of memory gB-T CD8+. CD103+ T cells isolated from the skin of C57/BL6 mice on day 30 p.i. with HSV KOS. 3 replicates of memory P14 CD8+ T cells isolated from gut of mice on day 60 p.i. with LCMV Armstrong. 3 replicates of memory gB-T CD8+ T cells from the lung of mice on day 30 p.i. with influenza WSN. 3 replicates of memory CD62L high CD8+ T cells from the spleen of mice on day 30 p.i. with HSV KOS. 3 replicates of memory CD62L low CD8+ T cells from the spleen of mice of day 30 p.i. with HSV KOS. 3 replicates of γδ-DETC isolated from the skin of C57/BL6 mice on day 30 p.i. with HSV KOS. 3 replicates of αβ-DETC from naive TCRδ-/- mice; and 3 replicates of naive gB-T CD8+ T cells from the spleen of naive gB-T transgenic mice.
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.
