{"title":"Guinea Pig and Mouse Models for Genital Herpes Infection.","authors":"Lauren M Hook, Harvey M Friedman, Sita Awasthi","doi":"10.1002/cpz1.332","DOIUrl":null,"url":null,"abstract":"<p><p>This article describes procedures for two preclinical animal models for genital herpes infection. The guinea pig model shares many features of genital herpes in humans, including a natural route of inoculation, self-limiting primary vulvovaginitis, spontaneous recurrences, symptomatic and subclinical shedding of HSV-2, and latent infection of the associated sensory ganglia (lumbosacral dorsal root ganglia, DRG). Many humoral and cytokine responses to HSV-2 infection in the guinea pig have been characterized; however, due to the limited availability of immunological reagents, assessments of cellular immune responses are lacking. In contrast, the mouse model has been important in assessing cellular immune responses to herpes infection. Both the mouse and guinea pig models have been extremely useful for evaluating preventative and immunotherapeutic approaches for controlling HSV infection and recurrent disease. In this article, we describe procedures for infecting guinea pigs and mice with HSV-2, scoring subsequent genital disease, and measuring replicating virus to confirm infection. We also provide detailed protocols for dissecting and isolating DRG (the site of HSV-2 latency), quantifying HSV-2 genomic copies in DRG, and assessing symptomatic and subclinical shedding of HSV-2 in the vagina. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Primary and recurrent genital herpes infection in the guinea pig model Support Protocol 1: Blood collection via lateral saphenous vein or by cardiac puncture after euthanasia Support Protocol 2: Dissection and isolation of dorsal root ganglia from guinea pigs Support Protocol 3: PCR amplification and quantification of HSV-2 genomic DNA from samples Basic Protocol 2: Primary genital herpes infection in the mouse model Alternate Protocol: Flank infection with HSV-2 in the mouse model Support Protocol 4: Dissection and isolation of mouse dorsal root ganglia.</p>","PeriodicalId":11174,"journal":{"name":"Current Protocols","volume":" ","pages":"e332"},"PeriodicalIF":0.0000,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8747063/pdf/nihms-1759051.pdf","citationCount":"5","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpz1.332","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 5
Abstract
This article describes procedures for two preclinical animal models for genital herpes infection. The guinea pig model shares many features of genital herpes in humans, including a natural route of inoculation, self-limiting primary vulvovaginitis, spontaneous recurrences, symptomatic and subclinical shedding of HSV-2, and latent infection of the associated sensory ganglia (lumbosacral dorsal root ganglia, DRG). Many humoral and cytokine responses to HSV-2 infection in the guinea pig have been characterized; however, due to the limited availability of immunological reagents, assessments of cellular immune responses are lacking. In contrast, the mouse model has been important in assessing cellular immune responses to herpes infection. Both the mouse and guinea pig models have been extremely useful for evaluating preventative and immunotherapeutic approaches for controlling HSV infection and recurrent disease. In this article, we describe procedures for infecting guinea pigs and mice with HSV-2, scoring subsequent genital disease, and measuring replicating virus to confirm infection. We also provide detailed protocols for dissecting and isolating DRG (the site of HSV-2 latency), quantifying HSV-2 genomic copies in DRG, and assessing symptomatic and subclinical shedding of HSV-2 in the vagina. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Primary and recurrent genital herpes infection in the guinea pig model Support Protocol 1: Blood collection via lateral saphenous vein or by cardiac puncture after euthanasia Support Protocol 2: Dissection and isolation of dorsal root ganglia from guinea pigs Support Protocol 3: PCR amplification and quantification of HSV-2 genomic DNA from samples Basic Protocol 2: Primary genital herpes infection in the mouse model Alternate Protocol: Flank infection with HSV-2 in the mouse model Support Protocol 4: Dissection and isolation of mouse dorsal root ganglia.
豚鼠和小鼠生殖器疱疹感染模型。
本文介绍了两种生殖器疱疹感染的临床前动物模型的制作过程。豚鼠模型具有人类生殖器疱疹的许多特征,包括自然接种途径,自限性原发性外阴阴道炎,自发性复发,HSV-2的症状和亚临床脱落,以及相关感觉神经节(腰骶背根神经节,DRG)的潜伏感染。豚鼠对HSV-2感染的许多体液和细胞因子反应已经被描述;然而,由于免疫试剂的可用性有限,缺乏对细胞免疫反应的评估。相比之下,小鼠模型在评估疱疹感染的细胞免疫反应方面很重要。小鼠和豚鼠模型对于评估控制HSV感染和复发性疾病的预防和免疫治疗方法都非常有用。在这篇文章中,我们描述了用HSV-2感染豚鼠和小鼠的程序,对随后的生殖器疾病进行评分,并测量复制病毒以确认感染。我们还提供了详细的解剖和分离DRG (HSV-2潜伏期部位)的方案,定量DRG中HSV-2的基因组拷贝,以及评估阴道中HSV-2的症状和亚临床脱落。©2021 Wiley期刊有限责任公司基本方案1:豚鼠模型的原发性和复发性生殖器疱疹感染支持方案1:通过侧隐静脉或安乐死后心脏穿刺采血支持方案2:解剖和分离豚鼠背根神经节支持方案3:从样本中提取HSV-2基因组DNA的PCR扩增和定量基本方案2:小鼠模型的原发性生殖器疱疹感染备用方案:支持方案4:小鼠背根神经节解剖分离。
本文章由计算机程序翻译,如有差异,请以英文原文为准。