Iversen Marie Beck, Paludan Søren Riis, Holm Christian Kanstrup
Department of Biomedicine, Faculty of Health Sciences, Aarhus University, Aarhus, Denmark.
Bio Protoc. 2017 Jul 5;7(13):e2383. doi: 10.21769/BioProtoc.2383.
The vaginal murine HSV-2 infection model is very useful in studying mucosal immunity against HSV-2 ( Overall , 1975 ; Renis , 1976 ; Parr and Parr, 2003). Histologically, the vagina of Depo-Provera-treated mice is lined by a single layer of mucus secreting columnar epithelial cells overlying two to three layers of proliferative cells. Even though this is morphologically different from the human vagina, it closely resembles the endocervical epithelium, which is thought to be the primary site of HSV-2 infection in women ( Parr , 1994 ; Kaushic , 2011). In the protocol presented here, mice are pre-treated with Depo-Provera before intra-vaginal inoculation with HSV-2. The virus replicates in the mucosal epithelium from where it spreads to and replicates in the CNS including the spinal cord, brain stem, cerebrum and cerebellum. Cytokine responses can be detected in vaginal washings using ELISA or in vaginal tissues using qPCR. Further, the recruitment of leukocytes to the vagina can be determined by flow cytometry. The model is suitable for research of both innate and adaptive immunity to HSV-2 infection.
阴道小鼠单纯疱疹病毒2型（HSV - 2）感染模型在研究针对HSV - 2的黏膜免疫方面非常有用（总体而言，1975年；雷尼斯，1976年；帕尔和帕尔，2003年）。从组织学上看，接受醋酸甲羟孕酮治疗的小鼠阴道由单层分泌黏液的柱状上皮细胞构成，其下有两到三层增殖细胞。尽管这在形态上与人类阴道不同，但它与子宫颈内膜上皮非常相似，子宫颈内膜上皮被认为是女性HSV - 2感染的主要部位（帕尔，1994年；考希克，2011年）。在本文介绍的实验方案中，小鼠在经阴道接种HSV - 2之前先用醋酸甲羟孕酮进行预处理。病毒在黏膜上皮中复制，然后从那里扩散到包括脊髓、脑干、大脑和小脑在内的中枢神经系统并在其中复制。可以使用酶联免疫吸附测定（ELISA）在阴道灌洗液中或使用实时定量聚合酶链反应（qPCR）在阴道组织中检测细胞因子反应。此外，可以通过流式细胞术确定白细胞向阴道的募集情况。该模型适用于对HSV - 2感染的固有免疫和适应性免疫的研究。
