Zhang Yongli, Zhang Hangjie, Ma Wenqiang, Liu Kefang, Zhao Min, Zhao Yingze, Lu Xuancheng, Zhang Fuping, Li Xiangdong, Gao George F, Liu William J
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention.
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention.
J Vis Exp. 2018 Oct 17(140):58110. doi: 10.3791/58110.
The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barré syndrome and damaging the testes. During an infection with the ZIKV, immune cells have been shown to infiltrate into the tissues. However, the cellular mechanisms that define the protection and/or immunopathogenesis of these immune cells during a ZIKV infection are still largely unknown. Herein, we describe methods to evaluate the virus-specific T-cell functionality in these immunoprivileged organs of ZIKV-infected mice. These methods include a) a ZIKV infection and vaccine inoculation in Ifnar1 mice; b) histopathology, immunofluorescence, and immunohistochemistry assays to detect the virus infection and inflammation in the brain, testes, and spleen; c) the preparation of a tetramer of ZIKV-derived T-cell epitopes; d) the detection of ZIKV-specific T cells in the monocytes isolated from the brain, testes, and spleen. Using these approaches, it is possible to detect the antigen-specific T cells that have infiltrated into the immunoprivileged organs and to evaluate the functions of these T cells during the infection: potential immune protection via virus clearance and/or immunopathogenesis to exacerbate the inflammation. These findings may also help to clarify the contribution of T cells induced by the immunization against ZIKV.
寨卡病毒（ZIKV）可在免疫赦免器官（如大脑和睾丸）中引发炎症，导致吉兰 - 巴雷综合征并损害睾丸。在寨卡病毒感染期间，免疫细胞已被证明会浸润到组织中。然而，在寨卡病毒感染期间，界定这些免疫细胞的保护作用和/或免疫发病机制的细胞机制仍大多未知。在此，我们描述了评估寨卡病毒感染小鼠这些免疫赦免器官中病毒特异性T细胞功能的方法。这些方法包括：a）在Ifnar1小鼠中进行寨卡病毒感染和疫苗接种；b）组织病理学、免疫荧光和免疫组织化学检测，以检测大脑、睾丸和脾脏中的病毒感染和炎症；c）制备寨卡病毒衍生的T细胞表位四聚体；d）检测从大脑、睾丸和脾脏分离的单核细胞中的寨卡病毒特异性T细胞。使用这些方法，可以检测浸润到免疫赦免器官中的抗原特异性T细胞，并评估这些T细胞在感染期间的功能：通过病毒清除实现潜在的免疫保护和/或通过免疫发病机制加剧炎症。这些发现也可能有助于阐明针对寨卡病毒免疫诱导的T细胞的作用。
