|
Author:
Date:
2013-11-20
[Abstract] Dengue virus infection usually courses as a benign self-limited fever, called dengue fever. However, on occasions it can progress to a life-threatening complication known as severe dengue (SD). A hallmark of SD is a sharp increase in vascular permeability. Secondary infections are considered a risk factor to develop SD, presumably through a mechanism called Antibody-Dependent Enhancement (ADE) of infection in cells with the capacity to bind antigen-antibody complexes, such as macrophages, and to trigger a subsequent aberrant cytokine response. The massive release of cytokine from macrophages has been postulated to cause changes in vascular permeability. The vascular permeability assay presented in this protocol is designed to assess whether any compound or cell-secreted product ...
[摘要] 登革热病毒感染通常称为良性自限发热，称为登革热。然而，偶尔，它可以进展为严重登革热（SD）的危及生命的并发症。 SD的标志是血管通透性的急剧增加。二次感染被认为是发展SD的风险因素，可能是通过在具有结合抗原 - 抗体复合物（例如巨噬细胞）能力的细胞中感染的感染的机制，称为抗体依赖性增强（ADE）引发随后的异常细胞因子应答。已经假定巨噬细胞中细胞因子的大量释放导致血管通透性的变化。在该方案中呈现的血管渗透性测定设计为评估来自患者的血清中存在的任何化合物或细胞分泌的产物或可溶性因子是否可以在小鼠中诱导血浆渗漏。在实验室中使用该测试以确定在存在促进性抗体的情况下由登革热病毒或登革热病毒感染的巨噬细胞在体外产生的细胞因子和可溶性因子是否能够在体内诱导血浆渗漏。在存在促进抗体的情况下，将巨噬细胞用登革病毒或登革热病毒感染48小时。此后，收集含有由巨噬细胞释放的细胞因子和可溶性因子的条件上清液，并腹膜内接种到CD-1小鼠中。在第一次接种后二十四小时，用伊文思蓝染料以第二剂量再次接种小鼠。再过24小时后，对小鼠实施安乐死，通过分光光度分析测定血液和肺中存在的伊文思蓝的量。该测定能够显示条件培养基在接种的动物中诱导血管通透性改变的能力的差异（Puerta-Guardo等人，2013）。
|