Guinea Pig and Mouse Models for Genital Herpes Infection.
Guinea Pig and Mouse Models for Genital Herpes Infection.
复制标题
DOI：
10.1002/cpz1.332
复制
发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Awasthi S
中科院分区：
文献类型：
--
作者：
Hook LM;Friedman HM;Awasthi S
本单元描述了生殖器疱疹感染的两个临床前动物模型的程序。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、自限性的原发外阴阴道炎、自发复发、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)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表征，然而，由于可获得的免疫试剂有限，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病毒感染的细胞免疫反应方面发挥了重要作用。小鼠和豚鼠模型对于评价预防和免疫治疗控制HSV感染和复发疾病的方法都是非常有用的。在本单元中，我们描述了用HSV-2感染豚鼠和小鼠、对随后的生殖器疾病进行评分以及测量复制病毒以确认感染的程序。我们还提供了详细的方案来解剖和分离DRG(HSV-2潜伏期的位置)，定量HSV-2在DRG中的基因组拷贝，以及评估HSV-2在阴道中的症状和亚临床脱落。豚鼠原发和复发性生殖器疱疹模型小鼠模型初发生殖器感染单纯疱疹病毒2型感染小鼠模型经隐静脉和心脏穿刺术(安乐死后)采血分离分离豚鼠背根神经节阴道拭子和背根节DNA提取及聚合酶链式反应从小鼠背根神经节提取HSV-2基因组DNA
This unit 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 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 to evaluate preventative and immunotherapeutic approaches for controlling HSV infection and recurrent disease. In this unit, 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), quantitating HSV-2 genomic copies in DRG, and assessing symptomatic and subclinical shedding of HSV-2 in the vagina. Primary and Recurrent Genital Herpes Infection in the Guinea Pig Model Primary Genital Infection in the Mouse Model Flank Infection with HSV-2 in the Mouse Model Blood Collection via the Lateral Saphenous Vein and Cardiac Puncture (following Euthanasia) Dissection and Isolation of DRG from Guinea Pigs DNA Isolation from Vaginal Swab Samples and DRG and PCR Amplification of HSV-2 genomic DNA Isolation of Mouse DRG