期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 58, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/3382
关键词
Immunology; Issue 58; Candida albicans; vaginitis; mouse; lumbar lymph nodes; vaginal tissues; vaginal lavage
资金
- NIAID, National Institute of Health [R01 AI32556]
- Louisiana Vaccine Center
- South Louisiana Institute for Infectious Disease Research
- Louisiana Board of Regents
Vulvovaginal candidiasis (VVC), caused by Candida species, is a fungal infection of the lower female genital tract that affects approximately 75% of otherwise healthy women during their reproductive years(18,32-34). Predisposing factors include antibiotic usage, uncontrolled diabetes and disturbance in reproductive hormone levels due to pregnancy, oral contraceptives or hormone replacement therapies(33,34). Recurrent VVC (RVVC), defined as three or more episodes per year, affects a separate 5 to 8% of women with no predisposing factors(33). An experimental mouse model of VVC has been established and used to study the pathogenesis and mucosal host response to Candida(3,4,11,16,17,19,21,25,37). This model has also been employed to test potential antifungal therapies in vivo(13,24). The model requires that the animals be maintained in a state of pseudoestrus for optimal Candida colonization/infection(6,14,23). Under such conditions, inoculated animals will have detectable vaginal fungal burden for weeks to months. Past studies show an extremely high parallel between the animal model and human infection relative to immunological and physiological properties(3,16,21). Differences, however, include a lack of Candida as normal vaginal flora and a neutral vaginal pH in the mice. Here, we demonstrate a series of key methods in the mouse vaginitis model that include vaginal inoculation, rapid collection of vaginal specimens, assessment of vaginal fungal burden, and tissue preparations for cellular extraction/isolation. This is followed by representative results for constituents of vaginal lavage fluid, fungal burden, and draining lymph node leukocyte yields. With the use of anesthetics, lavage samples can be collected at multiple time points on the same mice for longitudinal evaluation of infection/colonization. Furthermore, this model requires no immunosuppressive agents to initiate infection, allowing immunological studies under defined host conditions. Finally, the model and each technique introduced here could potentially give rise to use of the methodologies to examine other infectious diseases of the lower female genital tract (bacterial, parasitic, viral) and respective local or systemic host defenses.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据