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流式细胞术检测方案：用于定量分析单纯疱疹病毒-1 感染的小鼠角膜分离的上皮和基质中的免疫细胞。

Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea.

机构信息

Department of Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, MI 48201, USA.

Department of Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, MI 48201, USA.

出版信息

STAR Protoc. 2023 Mar 17;4(1):102056. doi: 10.1016/j.xpro.2023.102056. Epub 2023 Jan 25.

DOI:10.1016/j.xpro.2023.102056
PMID:36790766
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9898781/
Abstract

Existing flow cytometry approaches identify immune cells using the whole infected/inflamed cornea, which limits its ability to distinguish the immune cells infiltrating the corneal epithelium from the corneal stroma. Here, we present a protocol to analyze immune cells in the separated epithelium and stroma from naïve and herpes simplex virus-1 (HSV-1)-infected mouse corneas. We describe steps for viral infection, separation of corneal epithelium from stroma, preparation of a single-cell suspension of the individual epithelium and stroma, and flow cytometry assay.

摘要

现有的流式细胞术方法使用整个感染/炎症的角膜来识别免疫细胞，这限制了它区分浸润角膜上皮和基质的免疫细胞的能力。在这里，我们提出了一种从正常和单纯疱疹病毒 1（HSV-1）感染的小鼠角膜中分离的上皮和基质中分析免疫细胞的方案。我们描述了病毒感染、角膜上皮与基质分离、单个上皮和基质单细胞悬液制备以及流式细胞术检测的步骤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/280d87e0a537/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/c860c502be3f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/5ea343825e31/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/a479d5f41888/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/d4517026cf59/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/f94022a15e2d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/280d87e0a537/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/c860c502be3f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/5ea343825e31/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/a479d5f41888/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/d4517026cf59/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/f94022a15e2d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d76a/9898781/280d87e0a537/gr5.jpg

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Flow cytometry protocol to quantify immune cells in the separated epithelium and stroma of herpes simplex virus-1-infected mouse cornea.流式细胞术检测方案：用于定量分析单纯疱疹病毒-1 感染的小鼠角膜分离的上皮和基质中的免疫细胞。
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Comparison of an immortalized human corneal epithelial cell line with Vero cells in the isolation of Herpes simplex virus-1 for the laboratory diagnosis of Herpes simplex keratitis.在单纯疱疹性角膜炎实验室诊断中，用于分离单纯疱疹病毒1型的永生化人角膜上皮细胞系与Vero细胞的比较。
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引用本文的文献

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Differential homing of monocytes and neutrophils in the epithelial layer of HSV-1 infected cornea regulates viral dissemination and wound healing.单纯疱疹病毒1型感染角膜上皮层中单核细胞和中性粒细胞的差异性归巢调节病毒传播和伤口愈合。
Ocul Surf. 2025 Apr;36:69-82. doi: 10.1016/j.jtos.2025.01.002. Epub 2025 Jan 8.
2
Herpes stromal keratitis erodes the establishment of tissue-resident memory T cell pool in HSV-1 infected corneas.疱疹性基质性角膜炎破坏了单纯疱疹病毒1型感染角膜中组织驻留记忆T细胞池的建立。
Mucosal Immunol. 2025 Feb;18(1):188-204. doi: 10.1016/j.mucimm.2024.11.003. Epub 2024 Nov 22.
3
Foxp3 regulatory T cells reside within the corneal epithelium and co-localize with limbal stem cells.
Foxp3 调节性 T 细胞存在于角膜上皮内，并与角膜缘干细胞共定位。
Exp Eye Res. 2024 Dec;249:110123. doi: 10.1016/j.exer.2024.110123. Epub 2024 Oct 11.
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Novel characterization of CXCR4 expressing cells in uninfected and herpes simplex virus-1 infected corneas.在未感染和单纯疱疹病毒-1 感染的角膜中对 CXCR4 表达细胞的新特征描述。
Ocul Surf. 2023 Apr;28:99-107. doi: 10.1016/j.jtos.2023.02.006. Epub 2023 Feb 20.