Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Nat Protoc. 2010 Feb;5(2):239-46. doi: 10.1038/nprot.2009.227. Epub 2010 Jan 21.
The development of an HIV vaccine will require a more precise understanding of the immunological and virological underpinnings of HIV infection. Magnetofection, the process of magnetizing HIV by coupling it to ferrous nanoparticles and coordinating infection using a magnetic field, synchronizes the viral replication cycle at attachment while recapitulating the events of natural infection. Although spinoculation also concentrates virus onto target cells to increase infection, it does not synchronize infection. The synchronization of HIV infection in vitro facilitates the study of events in the viral replication cycle and the antiviral immune response on timelines previously impossible. Furthermore, by infecting a high percentage of cells in a short time frame, magnetofection increases the throughput of in vitro assays. Once a virus stock is generated, magnetofection of target cells is rapid, requiring only 1-2 h. Here we present a detailed protocol for this assay and review its applications for studying the immune response to HIV.
HIV 疫苗的开发需要更精确地了解 HIV 感染的免疫学和病毒学基础。磁转染是将 HIV 与亚铁纳米粒子偶联并使用磁场协调感染的过程,它在附着时使病毒复制周期同步,同时再现自然感染的事件。尽管自旋感染也将病毒浓缩到靶细胞上以增加感染,但它不能使感染同步。HIV 体外感染的同步化有助于研究病毒复制周期中的事件和抗病毒免疫反应,这在以前是不可能的。此外,通过在短时间内感染高比例的细胞,磁转染增加了体外分析的通量。一旦生成病毒库存,转染靶细胞的速度很快,仅需 1-2 小时。在这里,我们提出了该测定的详细方案,并回顾了其在研究 HIV 免疫反应中的应用。