Colony Stimulating Factor-1 Receptor Expressing Cells Infiltrating the Cornea Control Corneal Nerve Degeneration in Response to HSV-1 Infection

Chucair-Elliott, Ana J.; Gurung, Hem R.; Carr, Meghan M.; Carr, Daniel J.J.

doi:10.1167/iovs.17-22159

Cited by 18 publications

(18 citation statements)

References 66 publications

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“…Flow cytometric characterization of the cellular component that infiltrates the corneas of mice following reactivation revealed that the total number of inflammatory cells in wild-type mice significantly eclipsed that seen in their CD28KO counterparts. This observation was exactly as we predicted from the clinical scores and consistent with previous literature on this aspect of disease (3,(33)(34)(35). While B6-CD28KO mice did develop an inflammatory infiltrate consisting of CD4 ϩ T cells, CD8 ϩ T cells, neutrophils, and macrophages, their numbers were significantly less that that observed for wildtype B6 mice.…”

Section: Discussionsupporting

confidence: 92%

“…Recurrent infection causes an immunopathologic condition that leads to corneal inflammation and chronic damage. Previous studies have demonstrated that HSK requires intact T cells (17)(18)(19)(20) and an inflammatory infiltrate that consists of both neutrophils (15,33) and macrophages (34,35). Our present study extends those studies by indicating that CD28 costimulation of T cells is necessary for both primary and recurrent disease.…”

Section: Discussionsupporting

confidence: 83%

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CD28 Costimulation Is Required for Development of Herpetic Stromal Keratitis but Does Not Prevent Establishment of Latency

Yin

Baugnon

Potter

et al. 2019

J Virol

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Corneal infection with herpes simplex virus 1 (HSV-1) leads to infection of trigeminal ganglia (TG), typically followed by the establishment of latency in the infected neurons. When latency is disrupted, the virus reactivates and migrates back to the cornea, where it restimulates the immune response, leading to lesions in a disease called herpetic stromal keratitis (HSK). HSK requires T cell activation, as in the absence of T cells there is no disease. We decided to determine if CD28 costimulation of T cells was required in HSK. The results indicated that C57BL/6 CD28−/− and BALB/c CD28−/− mice failed to develop recurrent HSK, while their wild-type counterparts did. In order to better understand the dynamics of TG infection in these mice, we evaluated the amount of virus in infected TG and the number of individual neurons harboring latent virus. The results indicated that CD28−/− mice possessed significantly increased genome levels in their TG but many fewer LAT-positive cells than wild-type mice from day 7 to day 30 but that after day 30 these differences became nonsignificant. We next evaluated total and antigen-specific CD8+ T cells in TG. The results indicated that there were significantly fewer CD8 T cells in TG from day 10 to day 25 but that after that the differences were not significant. Taken together, these data suggest that CD28 costimulation is required for HSK but that while initial infection of TG is greater in CD28−/− mice, this begins to normalize with time and this normalization is concurrent with the delayed development of antigen-specific CD8+ T cells. IMPORTANCE We study the pathogenesis of herpes simplex virus-mediated corneal disease. T cells play a critical role both in disease and in the maintenance of latency in neurons. Consequently, the focus of this study was to evaluate the role that T cell costimulation plays both in corneal disease and in controlling the ability of the virus to maintain a stable infection of the ganglia that innervate the cornea. We demonstrate that in the absence of costimulation with CD28, corneal disease does not take place. However, this costimulation does not prevent the ability of CD8+ T cells to develop and, thus, control latent infection of neurons. We conclude from these studies that CD28 costimulation is required for corneal destructive immune responses but that CD8+ T cells develop over time and help to maintain latency.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 83%

CD28 Costimulation Is Required for Development of Herpetic Stromal Keratitis but Does Not Prevent Establishment of Latency

Yin

Baugnon

Potter

et al. 2019

J Virol

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“…The supernatants from tissue homogenates were assayed for protein content using a BCA protein method (#23225; ThermoFisher Scientific) and along with diluted plasma samples, used for protein analyses. Suspension array analyte concentrations were determined using a Bio-Rad Bio-Plex System Luminex 100 and Bio-Plex manager 5.0 (Bio-Rad Laboratories, Hercules, CA) 40 . Milliplex Map luminex-based assays were used to quantify the mouse inflammatory cytokines IL-6, TNFα, and INFɣ (#MCYTOMAG-70K; EMD Millipore, Billerica, MA).…”

Section: Methodsmentioning

confidence: 99%

Inducible cell-specific mouse models for paired epigenetic and transcriptomic studies of microglia and astroglia

Chucair-Elliott

Ocañas

Stanford

et al. 2019

Preprint

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23Epigenetic regulation of gene expression occurs in a cell type-specific manner. Current cell-type 24 specific neuroepigenetic studies rely on cell sorting methods that can alter cell phenotype and 25 introduce potential confounds. Here we demonstrate and validate a Nuclear Tagging and 26Translating Ribosome Affinity Purification (NuTRAP) approach for temporally controlled labeling 27 and isolation of ribosomes and nuclei, and thus RNA and DNA, from specific CNS cell types. 28Paired analysis of the transcriptome and DNA modifications in astrocytes and microglia 29 demonstrates differential usage of DNA methylation and hydroxymethylation in CG and non-CG 30 contexts that corresponds to cell type-specific gene expression. Application of this approach in 31 LPS treated mice uncovers microglia-specific transcriptome and epigenome changes in 32 inflammatory pathways that cannot be detected with tissue-level analysis. The NuTRAP model 33 and the validation approaches presented can be applied to any CNS cell type for which a cell 34 type-specific cre is available. 36Significant advances are being made in understanding the epigenome and its relationship with 37 gene expression in the brain 1-3 . However, the lack of approaches for paired analysis of DNA and 38RNA profiles at the cell type-specific level within the same animal is a significant limitation for the 39 field, given that epigenetic processes differ across CNS cell types at the level of chromatin 40 organization and DNA modifications 1,4 . Obtaining enriched cell populations by flow sorting 41 requires cell surface markers but these markers can change with experimental conditions and cell 42 sorting causes molecular, morphological, and functional changes, such as cell activation, that 43 could confound studies 3,5,6 . Single cell approaches 7 may overcome some of the challenges of 44 cell sorting but the scale of such studies, partial genomic coverage, restriction to only certain types 45 of endpoints, and continued potential for brain dissociation artifacts are limitations. 46This has led to development of transgenic labeling approaches to isolate RNA or DNA from 47 specific cell types. Ribosome labeling and RNA isolation methods, such as Translating Ribosome 48Affinity Purification (TRAP 8 ), and ribosome tagging (RiboTag 9 ), are gaining acceptance across 49 neuroscience studies examining the transcriptome. Similar approaches have been developed to 50 transgenically tag and allow isolation of nuclei and thus DNA (Isolation of Nuclei TAgged in 51 Specific Cell Types, INTACT) 10 . However, using separate transgenic mouse strains for DNA and 52RNA endpoints is a complicated and resource intensive approach. 53Here we describe an approach where Nuclear Tagging and Translating Ribosome Affinity 54Purification (NuTRAP) 11 is combined with well-established cell-specific inducible cre-recombinase 55 expressing systems 12,13 to perform paired transcriptomic and epigenomic analyses of specific 56 CNS cell types in a temporally controllable manner from a single mouse. ...

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“…For secondary chemiluminiscent detection, blots were incubated with corresponding HRP-conjugated secondary antibodies (GE Healthcare Bio-Sciences, Pittsburgh, PA). Imaging of the blots and analysis of band intensities were performed by conventional image analysis using a Kodak in vivo imaging system F Pro and Carestream MI SE 4.4 SE version software (Carestream Health, Inc., Rochester, NY) [(Chucair-Elliott et al 2017)]. Two-way ANOVA analyses were performed with the factors of sex and treatment.…”

Section: Protein Extraction and Western Blottingmentioning

confidence: 99%

Tamoxifen induction of Cre recombinase does not cause long-lasting or sexually divergent responses in the CNS epigenome or transcriptome: implications for the design of aging studies

et al. 2019

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The systemic delivery of tamoxifen (Tam) to activate inducible CreERT2-loxP transgenic mouse systems is now widely used in neuroscience studies. This critical technological advancement allows temporal control of DNA-cre recombination, avoidance of embryonically lethal phenotypes, and minimization of residual cell labeling encountered in constitutively active drivers. Despite its advantages, the use of Tam has the potential to cause long-lasting, uncharacterized side effects on the transcriptome and epigenome in the CNS, given its mixed estrogen receptor (ER) agonist/ antagonist actions. With the welcome focus on including both sexes in biomedical studies and efforts to understand sex differences, Tam administration could also cause sexually divergent responses that would confound studies. To examine these issues, epigenetic and transcriptomic profiles were compared in C57BL/6 J female and male hippocampus, cortex, and retina 1 month after a 5-day Tam treatment typical for cre induction, or vehicle control (sunflower seed oil). Cytosine methylation and hydroxymethylation levels, in both CG and non-CG contexts, were unchanged as

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Colony Stimulating Factor-1 Receptor Expressing Cells Infiltrating the Cornea Control Corneal Nerve Degeneration in Response to HSV-1 Infection

Cited by 18 publications

References 66 publications

CD28 Costimulation Is Required for Development of Herpetic Stromal Keratitis but Does Not Prevent Establishment of Latency

CD28 Costimulation Is Required for Development of Herpetic Stromal Keratitis but Does Not Prevent Establishment of Latency

Inducible cell-specific mouse models for paired epigenetic and transcriptomic studies of microglia and astroglia

Tamoxifen induction of Cre recombinase does not cause long-lasting or sexually divergent responses in the CNS epigenome or transcriptome: implications for the design of aging studies

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